Introduction
Acute kidney harm (AKI) is a extreme medical situation of abrupt kidney dysfunction related to structural damage and failure to keep the acid-base steadiness or remove waste merchandise.1 Many aetiological components are concerned in the pathophysiology of AKI, together with renal ischemia, nephrotoxic medication, endogenous toxins, or infections.2,3 It was reported that the lack of renal operate had been reversed in most survived sufferers. However, the deaths are nonetheless excessive (over 50%).4 To date, there aren’t any environment friendly therapies to stop or speed up the restoration from AKI. Hence, the seek for an efficient AKI remedy has gained a lot consideration.5
Intramuscular glycerol injection evokes AKI by induction of rhabdomyolysis (RM).3 This experimental mannequin is usually used for understanding the mechanisms of renal harm. The intensive breakdown of skeletal muscle mass generates a excessive quantity of myoglobin, surpassing plasma protein-binding capability, ensuing in glomerular filtration and tubular reabsorption of myoglobin. Myoglobin precipitation in the renal tubules aggravates tubular necrosis and obstruction with subsequent renal dysfunction.6,7 Consequent renal failure from RM-induced myoglobinuria represents about 10–40% of the whole AKI circumstances.8
Although the exact mechanisms implicated in RM-related AKI are unclear, current research have demonstrated that the pathogenesis of AKI encompasses the era of dangerous free radicals and the discharge of inflammatory mediators.1 Furthermore, tubular cell apoptosis is the dominant type of mobile loss of life in the AKI mannequin and is concerned in renal harm. In addition to apoptosis, necroptosis is a extremely regulated necrosis that will depend on phosphorylation of receptor-interacting protein kinase 3 (RIPK3) and strongly contributes to the pathogenesis of AKI.7,9 Former in vitro and in vivo investigations discovered that necroptosis inhibitors as necrostatin-1 or knockdown of RIP3 provoked noteworthy renoprotection in totally different AKI fashions.10–12
Abundance of experimental proof was reported that AKI-associated tissue damage could be ameliorated or averted by pure antioxidants supplementation through suppression of lipid peroxidation and scavenging of extra free radicals.1,3,4,13
Lycopene (LYC) is a naturally occurring carotenoid, richly discovered in tomato and tomato merchandise with potent antioxidant efficacy exceeding different antioxidants akin to β-carotene and α-tocopherol.14 Its outstanding antioxidant capability is attributed to quite a few double bonds allotted in its chemical construction.15 LYC has a powerful potential to quench singlet-oxygen and break peroxyl radicals.16 Earlier research have illustrated the antioxidant impact of LYC towards renal insult induced by cisplatin,17 gentamicin,18 methotrexate,15 colistin,19 and mercuric chloride16 toxicities. Also, LYC was proven to markedly relieve the renal irritation and apoptotic modifications in response to diabetic nephropathy and weight problems.20,21
Selenium, an important aspect, is a structural element of antioxidant enzymes as glutathione peroxidases and thioredoxin reductase.22 The use of conventional selenium compounds is proscribed as a result of its secure dose stage is slender, which restricts its utility. Therefore, selenium nanoparticles (SeNPs) space extra environment friendly and secure selenium type with excessive bioavailability and low toxicity.3 Preparation of SeNPs is often completed by chemical discount, utilizing a lowering agent and stabilizer. The use of stabilizers could hamper the organic utilization of SeNPs in addition to their chemical toxicity.23 Instead of the chemical methodology, nice consideration has been drawn lately to the eco-friendly or inexperienced synthesis of SeNPS utilizing biomolecules (plant proteins and polysaccharides) as a capping agent throughout processing.24–26 The eco-friendly strategy for SeNPs synthesis is healthier than different bodily and chemical strategies due to its low price, minimal hazard, and excessive efficiency.26 Recent research acknowledged that remedy with inexperienced synthesized SeNPs had noticeable antioxidant and anti-apoptotic actions.24,27
Based on our information, no analysis exists on the palliative impact of inexperienced synthesized SeNPs on the acute renal harm. Hence, this research was carried out to elucidate the potential renoprotective properties of LYC-capped SeNPs on renal damage relative to the only real remedy with both sodium selenite or LYC through evaluation of oxidative stress, irritation, apoptotic, and necroptotic signaling pathways.
Materials and Methods
Chemicals and Reagents
Sodium selenite and glycerol have been bought from Sigma Chemical Company, St. Louis, MO, USA. Lycopene was bought from Puritan’s Pride (Ronkonkoma, NY, USA). All different used chemical substances have been of excessive analytical grade.
Synthesis of Lycopene Coated-Selenium Nanoparticles (LYC-SeNPs)
Ten milliliters of 10 mM sodium selenite (Na2SeO3) was combined with 3.5 mg/mL LYC (10 mL) underneath magnetic stirring for twenty-four h. The obtained resolution of LYC-SeNPs was lyophilized by vacuum freeze dryer (Labconco Freezone 4.5 Liter Freeze Dry System, Marshall Scientific, Hampton NH, USA), and the obtained strong powder was utilized in the current research.
Characterization of the Prepared Nanoparticles (LYC-SeNPs)
The common diameter, dimension distribution, and floor costs of the LYC-SeNPs have been measured by the Zetasizer Nano ZS particle analyzer (Zetasizer Nano ZS90, Malvern Panalytical, UK). The absorbance of the LYC-SeNPs was measured at wavelengths ranging between 200 and 800 nm with 1 nm wavelength intervals utilizing UV–seen spectrophotometer (V-730 UV-visible spectrophotometer, JASCO, Japan). The crystalline dimension was characterised by X-ray diffraction (XRD; Malvern Panalytical, UK). Furthermore, transmission electron micrographs have been recorded utilizing a high-resolution transmission electron microscope (TEM; JEOL Ltd., Japan) outfitted with an electron diffraction sample. The molecular construction of LYC-SeNPs was analyzed utilizing Fourier Transform InfraRed spectroscopy (FTIR; PerkinElmer, USA).
Stability of Nanoparticles in Decreased or Increased pH
The stability of nanoparticles in decreased or elevated pH was studied by reducing or growing the pH regularly with 0.01 M HCl or 0.01 M NaOH, respectively, and the looks of LYC-SeNPs was noticed. The beginning pH was roughly 7.0 and the pH was decreased to 1.0 or elevated to 9.0 or till full dissolution or aggregation of nanoparticles. The pH was measured with the pH meter.
Animals and Ethics Statement
Wistar male albino rats at three months outdated (150–180 g) have been used for this research. They have been housed in the animal facility of the Zoology Department, Science of Faculty, Helwan University (Cairo, Egypt) underneath managed environmental circumstances (24 ± 2°C temperature, 50–60% relative humidity, and a daily light-dark cycle). Rats have been ate up customary rodent-chew diets (Harlan Laboratories Inc., Harlan, Indianapolis, IN; Teklad Diets T.2918) with free entry to water. Animal dealing with and all experimental procedures have been carried out following the rules of the Zoology Department, Faculty of Science, Helwan University (Approval quantity: HU2019/Z/AER0319-04).
AKI was induced in examined rats through intramuscular (IM) injection of fifty% glycerol (10 mL/kg) diluted in saline (0.9% NaCl) into the hind limbs with water restriction 24 hours earlier than the injection.28
Experimental Protocol
Rats have been allotted into 4 equal teams (7 rats per every) as follows:
Group 1 (CON): rats served as management and acquired intramuscular injection with physiological saline (0.9% NaCl).
Group 2 (AKI): rats acquired car for 14 days.
Group 3 (Na2SeO3+AKI): rats have been orally administered with Na2SeO3(0.5 mg/kg) for 14 days earlier than being subjected to AKI.
Group 4 (LYC+AKI): rats have been orally administered with LYC (10 mg/kg) for 14 days earlier than being subjected to AKI.
Group 5 (LYC-SeNPs+AKI): rats have been orally administered with LYC-SeNPs (0.5 mg/kg) for 14 days earlier than being subjected to AKI.
Twenty-four hours after the final remedy, all animals acquired an overdose of pentobarbital (100 mg/kg i.p.) and have been euthanized by decapitation. Blood samples have been collected for biochemical assessments. Kidneys have been then instantly eliminated and weighed. The proper kidney was used for performing the biochemical and molecular analyses, whereas the left one was examined for histopathological alterations.
Determination of Kidney Weight
The relative kidney weight was assessed primarily based on the next mathematical calculation:29
Relative kidney weight = (Left kidney weight/Body weight) x 100.
Preparation of Kidney Homogenate
A ten% (w/v) renal tissue was homogenized in 50 mM Tris–HCl (pH 7.4). The homogenate was centrifuged at 4°C for 10 min at 3,000 × g. The obtained supernatant was saved at −80°C for biochemical evaluation. The renal protein content material was measured utilizing bovine serum albumin as a reference protein in accordance to Lowry’s methodology.30
Determination of Intensity of Rhabdomyolysis
Lactate dehydrogenase (LDH) and creatine kinase (CK) have been evaluated utilizing kits provided by Randox/Laboratory, Crumlin, United Kingdom, following the producer’s instructions.
Assessment of Renal Function Biomarkers
Levels of urea, creatinine have been measured in serum samples by kits (Randox/Laboratory, Crumlin, United Kingdom) primarily based on the producer’s data.
Measurement of Kidney Injury Molecule-1 (Kim-1) and Neutrophil Gelatinase-Associated Lipocalin (NGAL)
The plasma ranges of Kim-1 (R&D Systems, Catalogue Number: AF3689) and NGAL (MyBioSource, Catalogue Number: MBS260195) have been analyzed by ELISA kits following the producer’s protocol.
Evaluation of Renal Non-Enzymatic Oxidative Stress Markers
Peroxidation of lipids was estimated by measured malondialdehyde (MDA).31 Further, nitric oxide (NO) content material in renal samples was measured by Griess reagent.32 Additionally, decreased glutathione (GSH) in renal tissue was evaluated.33
Assessment of Kidney Antioxidant Enzymatic Activities
Superoxide dismutase (SOD) and catalase (CAT) enzymatic actions have been measured in accordance to Nishikimi et al34 and Aebi,35 respectively. Moreover, glutathione peroxidase (GPx) and glutathione reductase (GR) enzymatic actions have been assessed in accordance to Paglia and Valentine36 and De Vega et al37 respectively.
Measurement of Inflammatory Biomarkers
Renal inflammatory response was assayed by quantification of tumor necrosis factor-α (TNF-α; Catalogue Number: EZRTNFA), interleukin-1β (IL-1β; Catalogue Number: RAB0278), and IL-6 (Catalogue Number: EZRIL6) utilizing kits provided by Merck Millipore (Toronto, Ontario, Canada) and Sigma-Aldrich (St. Louis, MO, USA), respectively, following the producers’ directions. The ranges of measured cytokines have been normalized by complete protein stage.
Assessment of the Renal Apoptotic Markers
Pro-apoptotic proteins (Cyt c, Bax and caspase-3), in addition to the anti-apoptotic protein (Bcl-2), have been measured by ELISA equipment (Cusabio, Wuhan, China) following the producer’s procedures. The catalogue Number for Cyt c: CSB-EL006328RA, Bax: CSB-EL002573RA, caspase-3: CSB-E08857r and Bcl-2: CSB-E08854r.
Quantitative Real-Time PCR
Extraction of complete RNA was completed from renal tissue by TRIzol reagent, adopted by cDNA synthesis utilizing RevertAid™ H Minus Reverse Transcriptase (Fermentas, Thermo Fisher Scientific Inc., Canada) as talked about in producer’s protocol. qrt-PCR was employed utilizing the QuantiFast SYBR Green RT-PCR equipment (Qiagen, Hilden, Germany). All reactions have been performed in duplicate utilizing the ViiA™ 7 System (Thermo Fisher Scientific, CA, USA). The PCR biking circumstances included preliminary denaturation at 95°C for 12 min, adopted by 40 cycles of denaturation at 94°C for 60s and annealing at 58°C for 60s, extension at 72 °C for 90s, then held for a remaining extension at 72°C for 10 min. The relative gene expression was decided between the totally different teams utilizing the ΔΔCt methodology.38 Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was used as a housekeeping gene. The primer sequences (Jena Bioscience [Jena, Germany]) for estimation of Nef2, Hmox-1, Nos2, and Ripk3 gene expressions are listed in Table 1.39
|
Table 1 Primer Sequences of Genes Analyzed in Real Time-PCR |
Light Microscopy and Histopathological Screening
The left kidneys from all studied teams have been mounted in 10% impartial buffered formalin for twenty-four h. Tissue samples have been dehydrated, embedded in paraffin, and eventually sectioned at 8 μm thick sections. Renal specimens have been stained with hematoxylin and eosin (H&E) and examined underneath a Nikon Eclipse E200-LED (Tokyo, Japan) microscope at 400× magnification.
Statistical Analysis
Obtained knowledge have been analyzed by one-way evaluation of variance (ANOVA) and Duncan’s post-hoc a number of assessments to assess the variations between the teams. The outcomes have been displayed as imply ± customary error of the imply (SEM). The stage of statistical significance was set at P-values decrease than 0.05 (P< 0.05).
Results
Physical and Chemical Characterization of the Prepared LYC-SeNPs
When including LYC to Na2SeO3, the colour modified from colorless to pink coloration (Figure 1A). SeNPs have been characterised with a median diameter of 129.3 nm (Figure 1B) and a imply zeta potential of −25.3 mV (Figure 1C). The UV–seen spectroscopic evaluation of the LYC-SeNPs revealed that the utmost absorption (λmax) was recorded at 353nm after in the future of preparation (knowledge not proven). Furthermore, the XRD sample end result confirmed a broader sample with none particular Braggs peaks. The obtained outcomes point out that LYC-SeNPs aren’t crystalline; slightly they’re extra amorphous (Figure 1D). The ready LYC-SeNPs-calculated crystalline dimension is <102 nm utilizing Scherrer’s equation. This amorphous character in settlement with the sooner research carried out with Withania somnifera39 and Bacillus sp.40 Moreover, TEM is employed to analyze the form of the LYC-SeNPs biosynthesized LYC. TEM picture of LYC-SeNPs revealed spherical particles throughout the diameter <130 nm. These particles are effectively distributed with no aggregation (Figure 1E). The FT-IR evaluation results of synthesized LYC-SeNPs is depicted in Figure 1F. A broad peak noticed at 3306.17 cm−1 corresponds to O–H stretch alcohols and phenols. The absorption peak at 2129.66 cm−1 corresponds to C–H stretch alkynes. The band at 1635.65 cm−1 is due to C–O uneven stretch carbon compounds. The absorption peak at 1039.05 cm−1is attributed to the C–N stretching of the amines. C–X stretching in alkyl halides causes a band at 602.98 cm−1. These outcomes reveal the presence of varied useful teams which may be accountable for each discount and stabilization of the SeNPs.
 |
Figure 1 Characterization of lycopene-coated selenium nanoparticles (LYC-SeNPs). (A) Color of LYC-SeNPs at 24 h. (B) Hydrodynamic diameter of LYC-SeNPs by Zetasizer. (C) Surface cost of LYC-SeNPs by Zeta potential. (D) XRD spectra of LYC-SeNPs. (E) Morphological form of LYC-SeNPs as noticed by TEM. (F) FT-IR spectra of LYC-SeNPs. |
The LYC-SeNPs nanoparticles have been steady in decreased pH until 1.0. However, the nanoparticles aggregated and/or precipitated as pH was elevated 11.3, however when the pH was decreased once more, the LYC-SeNPs have been resolved and the colour modified from colorless to pink coloration.
LYC-Capped SeNPs Improved RM-Related Biomarkers in the AKI Model
The sequences of LYC, Na2SeO3, and LYC-SeNPs supplementation on creatine kinase and LDH in glycerol-injected rats are proven in Figure 2. Marked will increase (P< 0.05) in RM indicators have been noticed in the AKI group associated to the management group. In distinction, when glycerol-injected rats have been co-administrated with LYC or Na2SeO3, substantial declines (P< 0.05) in RM parameters have been detected in contrast to the AKI group. Interestingly, the co-treatment with LYC-SeNPs resulted in marked declines (P> 0.05) in LDH stage in contrast to AKI and LYC administered group however not totally different from Na2SeO3 group. Creatine kinase stage displayed a pronounced lower (P<0.05) in the LYC-SeNPs group relative to the AKI and the handled teams with LYC and Na2SeO3. These outcomes urged that administration of LYC-SeNPs exerted higher safety on muscle fiber damage than the impact of both LYC or selenium alone.
 |
Figure 2 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on the rhabdomyolysis associated parameters in glycerol-induced AKI mannequin in rats. Data are expressed as imply ± SEM, n = 7. The statistical distinction between teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). |
Effect of LYC-Loaded SeNPs Administration on Kidney Weight and Function Markers
The mitigating impact of LYC-SeNPs on renal damage in glycerol-injected rats is illustrated in Figure 3. Significant increments (P< 0.05) have been detected in each kidney weight and relative kidney weight in the glycerol group relative to the management one, whereas their ranges confirmed notable decreases (P< 0.05) in the handled teams in contrast to the AKI group. No vital modifications (P> 0.05) have been observed upon evaluating the handled teams with LYC, Na2SeO3, or LYC-SeNPs in phrases of kidney weight or relative kidney weight. Regarding the serum kidney operate markers (urea and creatinine), markedly elevated ranges (P< 0.05) have been recorded in the AKI group with respect to the management group (Figure 3). However, their ranges exhibited marked decreases (P< 0.05) in teams that acquired Na2SeO3, LYC, or LYC-SeNPs in contrast to the AKI group. It is noteworthy that LYC-SeNPs co-administration evoked vital declines (P< 0.05) in creatinine ranges in contrast with the only real LYC or Na2SeO3 remedy.
 |
Figure 3 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on kidney weight and renal operate markers in glycerol-induced acute kidney harm. Data are expressed as imply ± SEM, n = 7. The statistical distinction between teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). |
Levels of NGAL and Kim-1 confirmed vital elevations (P< 0.05) following glycerol injection (Figure 3). Meanwhile, co-treatment with Na2SeO3, LYC, or LYC-SeNPs considerably decreased the concentrations of NGAL and Kim-1 in contrast to the untreated AKI group. Furthermore, the group co-treated with LYC-SeNPs had the bottom ranges (P< 0.05) of each NGAL and Kim-1 in contrast to the handled teams.
Augmentation of Renal Antioxidant Response After LYC-SeNPs Supplementation
The influence of various therapies on the renal tissue’s redox standing is represented in Figures 4 and 5. Marked oxidant/antioxidant imbalance was noticed in the AKI group, evidenced by vital elevations (P< 0.05) in MDA and NO ranges paralleled with marked depletion (P< 0.05) in GSH stage with respect to the management group. Moreover, outstanding suppressions (P< 0.05) in the antioxidant enzymatic actions of SOD, CAT, GPx, and GR after glycerol injection have been noticed. In distinction, co-treatment of the AKI group with Na2SeO3, LYC, or LYC-SeNPs improved the redox standing in the renal tissue through enhancement (P< 0.05) of the antioxidant protection members and GSH content material (P< 0.05) accompanied by diminished (P< 0.05) MDA and NO ranges when put next with the AKI untreated group. Co-administration with LYC-SeNPs resulted in vital GSH stage will increase and GPx accompanied by a decline in MDA stage with respect to the opposite handled teams. Remarkably, LYC-SeNPs supplementation reversed the oxidative stress in renal tissue as indicated by the restoration of antioxidant enzymatic actions (SOD, CAT, and GR) and GSH stage close to to their corresponding values in the management group.
 |
Figure 4 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on non-enzymatic antioxidant parameters [malondialdehyde (MDA), nitric oxide (NO), and glutathione (GSH)] ranges in glycerol-induced AKI in rats. Data are expressed as imply ± SEM, n = 7. The statistical distinction between teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). |
 |
Figure 5 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on antioxidant enzymatic actions in glycerol-induced acute kidney harm. Data are expressed as imply ± SEM, n = 7. The statistical distinction between the management and glycerol injected teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). Abbreviations: GPx, glutathione peroxidase, GR, glutathione reductase; SOD, superoxide dismutase, CAT, catalase. |
On the molecular foundation, marked downregulations (P< 0.05) have been detected in mRNA expression of nuclear issue, erythroid derived 2, like 2 (Nfe212), and heme oxygenase 1 (Hmox1) in the glycerol-induced AKI with respect to the management teams. In distinction, Na2SeO3 or LYC-treated rats had increased expression ranges (P< 0.05) of those antioxidant-related genes than rats with AKI. Lycopene capped SeNPs administration upregulated their expressions markedly (P< 0.05) with respect to the LYC or Na2SeO3 handled teams apart from the non-significant distinction in Hmox-1 expression stage in Na2SeO3 handled group (Figure 6).
 |
Figure 6 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on mRNA expression of Nfe212 and Hmox-1 in glycerol-induced AKI in rats. Data are expressed as imply ± SEM, n = 7. The obtained outcomes have been demonstrated because the imply ± SEM of triplicate experiments and have been referenced to Gapdh and represented as a fold change (log2 scale), with respect to mRNA ranges in the management group. The statistical distinction between teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). |
LYC-Coated SeNPs Alleviated Glycerol-Induced Renal Inflammation in the AKI Model
Compared to the renal tissue in the management group, vital will increase (P< 0.05) have been noticed in the IL-1β, IL-6, and TNF-α ranges related to upregulated Nos2 expression stage (P< 0.05) in the kidney tissue of glycerol-injected rats. Surprisingly, this irritation state of affairs in renal tissue was attenuated (P< 0.05) by the Na2SeO3, LYC, or LYC-SeNPs administration, as proven by the reversal in all parameters talked about above. Selenium nanoparticles loaded on LYC differed considerably (P< 0.05) from the LYC-treated group in phrases of IL-1β and TNF-α ranges (Figure 7).
 |
Figure 7 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on the degrees of inflammatory biomarkers (TNF-α, IL-1β and IL-6) and Nos2 mRNA expression in glycerol-induced AKI in rats. For ELISA outcomes, knowledge are expressed as imply ± SEM, n = 7. The statistical distinction between the management and glycerol injected teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). For qRT-PCR findings, the obtained outcomes have been demonstrated because the imply ± SEM of triplicate experiments and have been referenced to Gapdh and represented as a fold change (log2 scale), with respect to mRNA ranges in the management group. |
Amelioration of LYC-Conjugated SeNPs on Apoptosis and Necroptosis in Glycerol-Induced AKI
Glycerol-mediated AKI is related to vital induction of apoptotic and necroptotic pathways. As proven in Figure 8, vital elevations (P< 0.05) in pro-apoptotic proteins (caspase-3, Bax, and cyt-c) ranges accompanied by a marked decline (P< 0.05) in anti-apoptotic protein (Bcl-2) have been detected in kidneys with glycerol-induced tissue damage. Further, marked upregulation in mRNA expression of Ripk3, a necroptotic marker, in glycerol injected rats in contrast to the management group. In distinction, these parameters have been markedly decreased (P< 0.05) when glycerol-injected rats have been administered with Na2SeO3, LYC, or LYC-SeNPs in contrast to the AKI group. Notably, the appliance of LYC capped SeNPs remedy in the AKI group and reversed considerably (P< 0.05) the apoptotic damage in caspase-3, cyt-c, and Bcl-2 in contrast to rats administered with LYC or Na2SeO3 alone. These knowledge urged that when LYC coated SeNPs formulation was used, it supplied higher safety towards AKI-induced renal damage than both agent alone.
 |
Figure 8 The impact of lycopene coated selenium nanoparticles (LYC-SeNPs) on the degrees of apoptotic and necroptotic associated markers in glycerol-induced AKI in rats. For ELISA outcomes, knowledge are expressed as imply ± SEM, n = 7. The statistical distinction between teams was estimated utilizing Duncan’s post-hoc check at P < 0.05. Bars that don't share similar letters (superscripts) are considerably totally different from one another (p < 0.05). For qRT-PCR findings, the obtained outcomes have been demonstrated because the imply ± SEM of triplicate experiments and have been referenced to Gapdh and represented as a fold change (log2 scale), with respect to mRNA ranges in the management group. |
Effect of LYC-Loaded SeNPs Administration on Renal Histopathological Changes
Microscopic screening of stained renal sections from the management group confirmed regular tissue structure of the renal cortex and medulla (Figure 9). In distinction, sections from the glycerol-treated group revealed intensive tissue damage demonstrated by tubular dilation, vacuolation, necrosis, and particles accumulation in the tubular lumina. However, renal sections from rats handled with Na2SeO3 or LYC confirmed some enchancment of the renal histological modifications. Interestingly, LYC-SeNPs supplementation contributed to a notable enchancment in the renal tissue alteration.
 |
Figure 9 Histopathological alterations in the renal tissue following glycerol injection and totally different therapies. (A) Control, (B) AKI, (C) Na2SeO3+AKI, (D) LYC+AKI, and (E) LYC-SeNPs+AKI. Hematoxylin and eosin (H&E), scale bar= 50 μm. |
Discussion
Selenium nanoparticles have attracted intensive curiosity in the fields of nanomedicine owing to their distinctive organic actions. In specific, biogenic SeNPs synthesized by inexperienced nanotechnology characterize promising and secure selenium dietary supplements with low toxicity, excessive bioavailability, low price, and eco-friendly processing.41 In this regard, our research herein illustrated the potential of SeNPs synthesized utilizing LYC as a therapeutic strategy to alleviate AKI-related tissue damage in phrases of oxidative stress, irritation, apoptosis, and necroptosis.
Glycerol injection is usually used to induce AKI mannequin due to skeletal muscle degeneration and leakage of its content material into the circulation, which induces a myoglobinuric state comparable to medical RM. Consistent with earlier research, our outcomes revealed marked will increase in muscle damage markers (serum creatine kinase and LDH) in the glycerol-injected group in contrast to the management group.1,3,5 The launched merchandise have been reported to endure glomerular filtration inflicting intratubular obstruction, renal vasoconstriction, inflammatory response, and manufacturing of reactive oxygen species main to AKI.3,4 Renal impairment was evident in the AKI group as witnessed by outstanding elevations in serum urea and creatinine ranges related to elevated relative kidney weight as beforehand reported.5,7 In help, renal tissue histopathological alterations revealed tubular ischemia, vacuolation, and subsequent tubular necrosis and glomerular harm. Additionally, KIM-1, as a particular marker of renal harm positioned in broken renal epithelial cells in the proximal tubule, displayed a marked improve in the AKI group, which is in keeping with former research.7,42 NGAL, stress protein, belongs to the lipocalin superfamily and is taken into account an early diagnostic marker for AKI.43 In settlement with Sharawy et al43 a big increment was discovered in its stage in the glycerol-treated group that ensures renal damage.
Notably, LYC or Na2SeO3remedy demonstrated marked alleviation in biomarkers associated to RM and renal damage and renal pathological alterations supporting their capacities to enhance kidney operate and safeguard glycerol-induced damage. Earlier investigations indicated LYC remedy’s renoprotective impact towards nephrotoxic brokers.16–18 Additionally, Tsitsimpikou et al44 discovered that LYC effectively restored the degrees of LDH and creatine kinase ranges to nearly regular ranges in exercise-induced myocardial damage. The carotenoid LYC’s renoprotection could also be attributed to distinguished antioxidant and free radical scavenging capability and its cytoprotective efficacy towards proteins, lipids, and DNA oxidation.19 Similarly, sodium selenite co-treatment improved the renal capabilities and relieved the histological alterations induced by glycerol. These findings are attributed to its potent antioxidant capability and are in keeping with earlier outcomes.3,45,46 Interestingly, inexperienced synthesized SeNPs utilizing LYC supplied extra vital renoprotection than utilizing sodium selenite or LYC alone. Krishnan et al24 reported that SeNPs loaded with Spermacoce hispida exhibited many nephroprotective and hepatoprotective results towards acetaminophen toxicity in contrast to the only real remedy with SeNPs in rats. Similarly, Ramamurthy et al26 acknowledged that inexperienced synthesized SeNPs utilizing fenugreek seed extract notably inhibited the cell development in human breast most cancers cells in dose-dependent method. Further, LYC-rich nanoemulsions recorded the perfect safety of cardiomyoblast towards doxorubicin toxicity through its glorious cytoprotective properties.47
Multiple mechanisms are contributing to the pathogenesis of glycerol-induced AKI. The muscle damage induced by glycerol outcomes in the discharge of merchandise akin to myoglobin and heme derivates into the bloodstream. These merchandise have a catalytic iron metallic middle, which initiates free radicals manufacturing with consequent lipid peroxidation and oxidative stress in the proximal renal tubules.1,42 In the current research, renal tissue of the AKI mannequin exhibited marked elevations in MDA and NO ranges paralleled with a decline in GSH and suppression of SOD, CAT, GPx, and GR enzymatic actions. These findings agree with former authors.4,42 Supporting earlier stories, the present research revealed notable downregulations in mRNA expressions of each Nrf2 and HO-1 in renal tissue of the AKI group.4,48 Nrf2/HO-1 signaling pathway is reported to contribute to the oxidative harm in rhabdomyolysis-induced AKI.4,48 Nrf2 is a grasp transcriptional issue regulating a battery of genes encoding antioxidant proteins, like HO-1.49 HO-1 can convert heme to biliverdin and produce carbon monoxide with a cytoprotective position.1,49 Because of its significance in the catabolic technique of heme protein, it’s urged that HO-1 is included in the catabolism of myoglobin in broken renal tissues.48
LYC is taken into account the simplest antioxidant carotenoid, owing to single oxygen radical and different reactive species’ scavenging potential. During the quenching of1 O2, the LYC converts to a excessive energy-rich molecule, able to trapping different ROS, like OH, NO2, or peroxynitrite. However, additional trapping to different radicals leads to the oxidative breakdown of LYC.16,19 In our research, LYC recovered the oxidant/antioxidant imbalance in glycerol-induced renal harm markedly. These outcomes are in keeping with former stories.16,17,50 Furthermore, the molecular evaluation confirmed the antioxidant potential of LYC by notable stimulation for the Nrf2/HO-1 signaling pathway, which is concomitant with Dai et al.19
Owing to its important incorporation into selenium-dependent enzymes as GPx and thioredoxin reductases, selenium can adequately deal with the free radicals and keep the redox steadiness contained in the cells.3,46 Our findings revealed marked enchancment in the mobile antioxidant capability in renal tissue of sodium selenite-treated group, which is in settlement with earlier authors.46 Using LYC for the synthesis of SeNPs in our research supplied distinguished antioxidant actions in contrast to the co-treatment with LYC or sodium selenite. Zhang et al25 acknowledged that the SeNPs capped with Lycium barbarum polysaccharides had highly effective antioxidant potential, they usually have a protecting impact towards H2O2-induced cell loss of life in the PC-12 cell line. It is assumed that the helpful impact of LYC-SeNPs is endorsed for the synergistic results that may be generated between plant polysaccharides and SeNPs in addition to the elevated bioavailability and bioactivity.27
Activation of pro-inflammatory pathways and renal irritation is a typical pathophysiological characteristic of rhabdomyolysis-induced AKI, which can exacerbate tubular epithelium damage and fibrosis.3,6,7 In our research, vital increments have been detected in the degrees of IL-1β, TNF-α and IL-6, in addition to the fold change of Nos2 in the glycerol-injected group. Such a powerful inflammatory response in the AKI mannequin could also be attributed to extra ROS manufacturing and activation of nuclear issue kappa-B (NF-κB), which controls the expression of iNOS and pro-inflammatory cytokines.2 Additionally, iNOS triggers the era of reactive nitrogen species that additionally work together with superoxide anions to produce peroxynitrite radical and improve the manufacturing of pro-inflammatory cytokines.43
Marked lessening of the renal irritation was noticed in LYC or sodium selenite co-treated rats as indicated by lowered IL-1β, IL-6, and TNF-α and downregulated iNOS expression, in keeping with the findings of earlier research.20,21 The anti-inflammatory motion of LYC refers to the inhibition of nuclear issue kappa B and the manufacturing of pro-inflammatory mediators. Likewise, earlier stories acknowledged that elenium exerts its anti-inflammatory exercise through suppressingNF-κB,3 iNOS, and COX-2.49 Nanoformulation of selenium primarily based on LYC achieved a marked discount in TNF-α stage than the person LYC or selenium did, indicating the optimized bioavailability of SeNPs. Quagliariello et al47 acknowledged that lycopene-rich nanoemulsion probably lessened the inflammatory cytokines and NO in cardiomyocytes uncovered to doxorubicin. Also, SeNPs coated with Ulva lactuca polysaccharide markedly diminished the irritation in acute colitis.51
In addition to the mentioned pivotal position of oxidative stress and irritation in renal harm, apoptosis and necroptosis of renal tubular epithelial cells are additionally essential in glycerol-induced AKI’s pathophysiology. In our research, noteworthy will increase have been recorded in renal Bax, caspase-3, and Cyt-c ranges related to a marked decline in Bcl2 in the glycerol-injected group indicating a pronounced shift in the direction of the path of apoptosis. Myoglobin-induced oxidative stress alters the mitochondrial permeability with the following launch of cytochrome-c and caspase activation.52 These findings have been in line with former research.3,5,7 Necroptosis, a sort of regulated necrosis, is a caspase-independent loss of life pathway concerned in glycerol induced AKI.52 Also, renal harm induced by glycerol was accompanied by a marked upregulation in Ripk3 expression. It was previously documented that TNF-α has an important position in triggering the necroptotic signaling pathway through interplay with TNF receptor 1 (TNFR1) and phosphorylation of RIPK3 by RIPK1. Phosphorylated RIPK1 and RIPK3 activate combined lineage kinase domain-like pseudokinase (MLKL), which additionally management necroptosis with RIPK.5 This end result got here in line with earlier authors.5,7,52
Co-treatment with LYC evoked marked anti-apoptotic and anti-necroptotic actions in the glycerol-injected group, which is in settlement with earlier research.53,54 The vital cytoprotective impact of LYC in renal tissue could refer to its antioxidant potential, activation of the Nrf2 pathway paralleled with suppression of the NF-κB, and caspase/Bcl2 apoptotic pathways.19 It was reported that administration of necrostatin-1, a RIPK1 inhibitor, decreased creatinine plasma ranges and tubular necrosis.55 Accordingly, the anti-necroptotic impact of LYC or selenium is assumed to be associated to their inhibitory motion on Ripk3. In help of former outcomes,27,45 LYC-SeNPs exerted an anti-apoptotic impact in contrast to LYC or selenium solely due to suppression of caspases.
Conclusion
Taken collectively, biogenic SeNPs synthesized by LYC exhibited appreciable nephroprotective exercise towards AKI-caused tissue damage in rat fashions. The attainable mechanisms of LYC-SeNPs results could also be attributed to free radical scavenging potential with the enhancement of the endogenous antioxidant system, attenuating the renal irritation, anti-apoptotic and anti-necroptotic actions. These findings urged that the synthesis of SeNPs by LYC is a promising selenium complement, which is a novel therapeutic technique in the amelioration of AKI-related tissue damage.
Funding
This work was supported by Taif University Researchers Supporting Program (Project quantity: TURSP-2020/151), Taif University, Saudi Arabia.
Disclosure
The authors declare no conflicts of curiosity in this work.
References
1. Yin M, Jiang N, Guo L, et al. Oleuropein suppresses oxidative, inflammatory, and apoptotic responses following glycerol-induced acute kidney harm in rats. Life Sci. 2019;232:116634. doi:10.1016/j.lfs.2019.116634
2. Makris Okay, Spanou L. Acute kidney harm: definition, pathophysiology and medical phenotypes. Clin Biochem Rev. 2016;37(2):85.
3. AlBasher G, Alfarraj S, Alarifi S, et al. Nephroprotective position of selenium nanoparticles towards glycerol-induced acute kidney harm in rats. Biol Trace Elem Res. 2020;194(2):444–454. doi:10.1007/s12011-019-01793-5
4. Wu J, Pan X, Fu H, et al. Effect of curcumin on glycerol-induced acute kidney harm in rats. Sci Rep. 2017;7(1):1–11. doi:10.1038/s41598-016-0028-x
5. Abd-Ellatif RN, Hegab II, Atef MM, Sadek MT, Hafez YM. Diacerein protects towards glycerol-induced acute kidney harm: modulating oxidative stress, irritation, apoptosis and necroptosis. Chem Biol Interact. 2019;306:47–53. doi:10.1016/j.cbi.2019.04.008
6. Reis NG, Francescato HDC, de Almeida LF, da Silva CGA, Costa RS, Coimbra TM. Protective impact of calcitriol on rhabdomyolysis-induced acute kidney harm in rats. Sci Rep. 2019;9(1):1–10. doi:10.1038/s41598-019-43564-1
7. Li YF, Xu BY, An R, et al. Protective impact of anisodamine in rats with glycerol-induced acute kidney harm. BMC Nephrol. 2019;20(1):223. doi:10.1186/s12882-019-1394-y
8. Al Asmari AK, Al Sadoon KT, Obaid AA, Yesunayagam D, Tariq M. Protective impact of quinacrine towards glycerol-induced acute kidney harm in rats. BMC Nephrol. 2017;18(1):41. doi:10.1186/s12882-017-0450-8
9. Linkermann A, Chen G, Dong G, Kunzendorf U, Krautwald S, Dong Z. Regulated cell loss of life in AKI. J Am Soc Nephrol. 2014;25(12):2689–2701. doi:10.1681/ASN.2014030262
10. Wang S, Zhang C, Hu L, Yang C. Necroptosis in acute kidney harm: a shedding mild. Cell Death Dis. 2016;7(3):e2125. doi:10.1038/cddis.2016.37
11. Zhang L, Jiang F, Chen Y, et al. Necrostatin-1 attenuates ischemia harm induced cell loss of life in rat tubular cell line NRK-52E by decreased Drp1 expression. Int J Mol Sci. 2013;14(12):24742–24754. doi:10.3390/ijms141224742
12. Linkermann A, Bräsen JH, Darding M, et al. Two unbiased pathways of regulated necrosis mediate ischemia–reperfusion harm. Proc Natl Acad Sci. 2013;110(29):12024–12029. doi:10.1073/pnas.1305538110
13. Guo LP, Liu SX, Yang Q, et al. Effect of thymoquinone on acute kidney harm induced by sepsis in BALB/c mice. Biomed Res Int. 2020;2020:1594726.
14. Ramadan SS, Almeer R, Albasher G, Abdel Moneim AE. Lycopene mitigates arsenic-induced nephrotoxicity with activation of the Nrf2 pathway in mice. Toxin Rev. 2021;1–11. doi:10.1080/15569543.2021.1891938
15. Stojiljkovic N, Ilic S, Jakovljevic V, et al. The encapsulation of lycopene in nanoliposomes enhances its protecting potential in methotrexate-induced kidney harm mannequin. Oxid Med Cell Longev. 2018;2018:1–11. doi:10.1155/2018/2627917
16. Augusti PR, Conterato GM, Somacal S, et al. Effect of lycopene on nephrotoxicity induced by mercuric chloride in rats. Basic Clin Pharmacol Toxicol. 2007;100(6):398–402. doi:10.1111/j.1742-7843.2007.00067.x
17. Erman F, Tuzcu M, Orhan C, Sahin N, Sahin Okay. Effect of lycopene towards cisplatin-induced acute renal harm in rats: natural anion and cation transporters analysis. Biol Trace Elem Res. 2014;158(1):90–95. doi:10.1007/s12011-014-9914-x
18. Bayomy NA, Elbakary RH, Ibrahim MAA, Abdelaziz EZ. Effect of lycopene and rosmarinic acid on gentamicin induced renal cortical oxidative stress, apoptosis, and autophagy in grownup male albino rat. Anat Rec. 2017;300(6):1137–1149. doi:10.1002/ar.23525
19. Dai C, Tang S, Deng S, et al. Lycopene attenuates colistin-induced nephrotoxicity in mice through activation of the Nrf2/HO-1 pathway. Antimicrob Agents Chemother. 2015;59(1):579–585. doi:10.1128/AAC.03925-14
20. Guo Y, Liu Y, Wang Y. Beneficial impact of lycopene on anti-diabetic nephropathy by diminishing inflammatory response and oxidative stress. Food Funct. 2015;6(4):1150–1156. doi:10.1039/C5FO00004A
21. Pierine DT, Navarro ME, Minatel IO, et al. Lycopene supplementation reduces TNF-α through RAGE in the kidney of overweight rats. Nutr Diabetes. 2014;4(11):e142. doi:10.1038/nutd.2014.39
22. Liu L, Liu C, Hou L, et al. Protection towards ischemia/reperfusion‑induced renal harm by co‑remedy with erythropoietin and sodium selenite. Mol Med Rep. 2015;12(6):7933–7940. doi:10.3892/mmr.2015.4426
23. Sowndarya P, Ramkumar G, Shivakumar M. Green synthesis of selenium nanoparticles conjugated Clausena dentata plant leaf extract and their insecticidal potential towards mosquito vectors. Artif Cells, Nanomed Biotechnol. 2017;45(8):1490–1495. doi:10.1080/21691401.2016.1252383
24. Krishnan V, Loganathan C, Thayumanavan P. Green synthesized selenium nanoparticles utilizing Spermacoce hispida as provider of s-allyl glutathione: to accomplish hepatoprotective and nephroprotective exercise towards acetaminophen toxicity. Artif Cells, Nanomed Biotechnol. 2019;47(1):56–63. doi:10.1080/21691401.2018.1543192
25. Zhang W, Zhang J, Ding D, et al. Synthesis and antioxidant properties of Lycium barbarum polysaccharides capped selenium nanoparticles utilizing tea extract. Artif Cells, Nanomed Biotechnol. 2018;46(7):1463–1470. doi:10.1080/21691401.2017.1373657
26. Ramamurthy C, Sampath KS, Arunkumar P, et al. Green synthesis and characterization of selenium nanoparticles and its augmented cytotoxicity with doxorubicin on most cancers cells. Bioprocess Biosyst Eng. 2013;36(8):1131–1139. doi:10.1007/s00449-012-0867-1
27. Wang L, Li C, Huang Q, Fu X. Biofunctionalization of selenium nanoparticles with a polysaccharide from Rosa roxburghii fruit and their protecting impact towards H(2)O(2)-induced apoptosis in INS-1 cells. Food Funct. 2019;10(2):539–553. doi:10.1039/C8FO01958D
28. Kim JH, Lee SS, Jung MH, et al. N-acetylcysteine attenuates glycerol-induced acute kidney harm by regulating MAPKs and Bcl-2 household proteins. Nephrol Dial Transplant. 2010;25(5):1435–1443. doi:10.1093/ndt/gfp659
29. Almeer RS, AlBasher GI, Alarifi S, Alkahtani S, Ali D, Abdel Moneim AE. Royal jelly attenuates cadmium-induced nephrotoxicity in male mice. Sci Rep. 2019;9(1):5825. doi:10.1038/s41598-019-42368-7
30. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265–275. doi:10.1016/S0021-9258(19)52451-6
31. Ohkawa H, Ohishi N, Yagi Okay. Assay for lipid peroxides in animal tissues by thiobarbituric acid response. Anal Biochem. 1979;95(2):351–358. doi:10.1016/0003-2697(79)90738-3
32. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in organic fluids. Anal Biochem. 1982;126(1):131–138. doi:10.1016/0003-2697(82)90118-X
33. Ellman GL. Tissue sulfhydryl teams. Arch Biochem Biophys. 1959;82(1):70–77. doi:10.1016/0003-9861(59)90090-6
34. Nishikimi M, Appaji N, Yagi Okay. The incidence of superoxide anion in the response of decreased phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun. 1972;46(2):849–854. doi:10.1016/S0006-291X(72)80218-3
35. Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121–126.
36. Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med. 1967;70(1):158–169.
37. De Vega L, Fernandez RP, Mateo MC, Bustamante JB, Herrero AM, Munguira EB. Glutathione willpower and a research of the exercise of glutathione-peroxidase, glutathione-transferase, and glutathione-reductase in renal transplants. Ren Fail. 2002;24(4):421–432. doi:10.1081/JDI-120006769
38. Livak KJ, Schmittgen TD. Analysis of relative gene expression knowledge utilizing real-time quantitative PCR and the two(-Delta Delta C(T)) methodology. Methods. 2001;25(4):402–408. doi:10.1006/meth.2001.1262
39. Alagesan V, Venugopal S. Green synthesis of selenium nanoparticle utilizing leaves extract of withania somnifera and its organic purposes and photocatalytic actions. BioNanoScience. 2019;9(1):105–116. doi:10.1007/s12668-018-0566-8
40. Rajkumar Okay, Mvs S, Koganti S, Burgula S. Selenium nanoparticles synthesized utilizing pseudomonas stutzeri (MH191156) present antiproliferative and anti-angiogenic exercise towards cervical most cancers cells. Int J Nanomedicine. 2020;15:4523–4540. doi:10.2147/IJN.S247426
41. Qiao L, Dou X, Yan S, Zhang B, Xu C. Biogenic selenium nanoparticles synthesized by Lactobacillus casei ATCC 393 alleviate diquat-induced intestinal barrier dysfunction in C57BL/6 mice by their antioxidant exercise. Food Funct. 2020;11(4):3020–3031. doi:10.1039/D0FO00132E
42. Siddiqui RA, Simjee SU, Kabir N, Ateeq M, Shah MR, Hussain SS. N-(2-hydroxyphenyl)acetamide and its gold nanoparticle conjugation stop glycerol-induced acute kidney harm by attenuating irritation and oxidative harm in mice. Mol Cell Biochem. 2019;450(1–2):43–52. doi:10.1007/s11010-018-3371-3
43. Sharawy MH, Abdelrahman RS, El-Kashef DH. Agmatine attenuates rhabdomyolysis-induced acute kidney harm in rats in a dose dependent method. Life Sci. 2018;208:79–86. doi:10.1016/j.lfs.2018.07.019
44. Tsitsimpikou C, Kioukia-Fougia N, Tsarouhas Okay, et al. Administration of tomato juice ameliorates lactate dehydrogenase and creatinine kinase responses to anaerobic coaching. Food Chem Toxicol. 2013;61:9–13. doi:10.1016/j.fct.2012.12.023
45. Li X, Wang Q, Deng G, et al. Porous [email protected] SiO2 nanospheres attenuate cisplatin-induced acute kidney harm through activation of Sirt1. Toxicol Appl Pharmacol. 2019;380:114704. doi:10.1016/j.taap.2019.114704
46. Randjelovic P, Veljkovic S, Stojiljkovic N, et al. Protective impact of selenium on gentamicin-induced oxidative stress and nephrotoxicity in rats. Drug Chem Toxicol. 2012;35(2):141–148. doi:10.3109/01480545.2011.589446
47. Quagliariello V, Vecchione R, Coppola C, et al. Cardioprotective results of nanoemulsions loaded with anti-inflammatory nutraceuticals towards doxorubicin-induced cardiotoxicity. Nutrients. 2018;10(9):1304. doi:10.3390/nu10091304
48. Zhao W, Huang X, Zhang L, et al. Penehyclidine hydrochloride pretreatment ameliorates rhabdomyolysis-induced AKI by activating the Nrf2/HO-1 pathway and assuaging endoplasmic reticulum stress in rats. PLoS One. 2016;11(3):e0151158. doi:10.1371/journal.pone.0151158
49. Albarakati AJA, Baty RS, Aljoudi AM. et al. Luteolin protects towards lead acetate-induced nephrotoxicity by antioxidant, anti-inflammatory, anti-apoptotic, and Nrf2/HO-1 signaling pathways. Mol Biol Rep. 2020:1–13. doi:10.1007/s11033-019-04608-x
50. Yu Okay, Zhang J, Cao Z, et al. Lycopene attenuates AFB 1-induced renal harm with the activation of the Nrf2 antioxidant signaling pathway in mice. Food Funct. 2018;9(12):6427–6434. doi:10.1039/C8FO01301B
51. Zhu C, Zhang S, Song C, et al. Selenium nanoparticles embellished with Ulva lactuca polysaccharide probably attenuate colitis by inhibiting NF-κB mediated hyper irritation. J Nanobiotechnology. 2017;15(1):1–15. doi:10.1186/s12951-017-0252-y
52. Guerrero-Hue M, García-Caballero C, Palomino-Antolín A, et al. Curcumin reduces renal damage related to rhabdomyolysis by reducing ferroptosis-mediated cell loss of life. FASEB J. 2019;33(8):8961–8975. doi:10.1096/fj.201900077R
53. Dogukan A, Tuzcu M, Agca CA, et al. A tomato lycopene advanced protects the kidney from cisplatin-induced harm through affecting oxidative stress in addition to Bax, Bcl-2, and HSPs expression. Nutr Cancer. 2011;63(3):427–434. doi:10.1080/01635581.2011.535958
54. Çevik Ö, Oba R, Macit Ç, et al. Lycopene inhibits caspase-3 exercise and reduces oxidative organ damage in a rat mannequin of thermal harm. Burns. 2012;38(6):861–871. doi:10.1016/j.burns.2012.01.006
55. Homsi E, Andreazzi DD, Faria JBLD, Janino P. TNF-α-mediated cardiorenal harm after rhabdomyolysis in rats. Am J Physiol Renal Physiol. 2015;308(11):F1259–F1267. doi:10.1152/ajprenal.00311.2014
