Rat Kidney Research Articles

Effects of organophosphates on precision-cut kidney slices.

Organophosphate (OP) poisoning, both accidental and with suicidal intent, is a global medical challenge. While the primary toxicity of these pesticides is based on the inhibition of acetylcholinesterase (AChE), case reports describe patients developing OP-mediated renal insufficiency. We set out to investigate possible pathomechanisms utilizing rat precision-cut kidney slices (PCKS). Depending on the method of investigation, PCKS were observed for a maximum of 10 days. PCKS exposed to OP compounds (malaoxon, malathion, paraoxon, parathion) showed a dose-dependent loss of viability and a reduction of total protein content over the course of 10 days. A concentration of 500 µM OP showed the most differences between OP compounds. After two days of incubation parathion showed a significantly lower level of viability than malathion. The respective effects of paraoxon and malaoxon were not significantly different from control. However, effects of OP were only observed in concentrations exceeding those that were needed to achieve significant AChE inhibition in rat kidney tissue. In addition, we observed histological changes, without inducing LDH leakage. Overall, results suggest that OP exert effects in kidney tissue, that exceed those expected from the sole inhibition of AChE and vary between compounds. Without signs of necrosis, findings call for studies that address other possible pathomechanisms including inflammatory response, oxidative stress or activation of apoptosis to further understand nephrotoxicity of OP compounds. Monitoring oxon concentration over time, we demonstrated reduced enzyme inhibiting properties in the presence of PCKS, suggesting interactions between OP compound and kidney tissue.

Magnitude of Urine Albumin and KIM-1 Changes Can be Used to Differentiate Glomerular Injury From Tubular Injury in Rats.

Emerging urinary kidney safety biomarkers have been evaluated in recent years and have been shown to be superior to the serum parameters blood urea nitrogen (BUN) and creatinine (sCr) for monitoring kidney injury in the proximal tubule. However, their potential application in differentiating the location of the initial kidney injury (eg, glomerulus vs tubule) has not been fully explored. Here, we assessed the performance of two algorithms that were constructed using either an empirical or a mathematical model to predict the site of kidney injury using a data set consisting of 22 rat kidney toxicity studies with known urine biomarker and histopathologic outcomes. Two kidney safety biomarkers used in both models, kidney injury molecule 1 (KIM-1) and albumin (ALB), were the best performers to differentiate glomerular injury from tubular injury. The performance of algorithms using these two biomarkers against the gold standard of kidney histopathologic examination showed high sensitivity in differentiating the location of the kidney damage to either the glomerulus or the proximal tubules. These data support the exploration of such an approach for use in clinical settings, leveraging urinary biomarker data to aid in the diagnosis of either glomerular or tubular injury where histopathologic assessments are not conducted.

Effects of short‐ and long‐term use of propolis extracts on liver and kidney in rats

AbstractPropolis is widely used as a supplementary food product for its health benefits. The aim of this study was to determine the effects of commercial propolis extracts on the liver and kidney. Propolis extracts (250 mg/kgbw/day) were administered orally to adult male Wistar albino rats in solvents of ethanol, propylene glycol, water, and olive oil. Liver enzyme levels were determined biochemically in blood samples, and histopathological examinations were performed on the liver. Damage rate in both kidney tissue in the propolis‐ethanol extract group increased significantly compared with the other groups after 30 and 90 days of application (p < .05). According to the results, ethanol, used as a common solvent in propolis products, may adversely affect the liver in long‐term use. The data indicate that propolis‐olive oil extract may be an essential alternative due to its effective and reliable properties.

Evaluation of the feeding safety of Moringa (Moringa oleifera L.) in the Sprague Dawley rat

This study aimed to evaluate the safety of Moringa by comparing the effects of different gavage doses of Moringa. The general behavior, body weight, food intake, blood indexes, serum biochemical indexes, and histopathology of rats were used to determine the safety threshold and to provide a reference for the further development and use of Moringa as animal feed. 40 Sprague Dawley rats were selected and given transoral gavage for 28 consecutive days. The T1, T2 and T3 groups were observed for general behavior, body weight, and food intake. Blood and serum biochemical indices were quantified, and histopathology was performed to evaluate the effect and safety of Moringa. The results of the toxicological test showed that (1) Only T1 groups experienced diarrhea. (2) The body weight and food intake of rats in each group were normal compared with the control group. (3) The hematological and serum biochemical indices of rats in the T1 group were significantly different from those of CK but were in the normal range; (4) The results of microscopic examination of the heart, liver, spleen, lung, and kidney of rats in each group were normal, but inflammation occurred in stomach and jejunum of rats in the T1 group, but not in the ileum. The gastrointestinal tract of rats in the T2 and T3 groups were normal. (5) No abnormal death occurred in any of the treatment groups.The results of this study revealed that gavage of Moringa homogenate at a dose of 6 g/kg BW can cause diarrhea in rats. Although there is no pathological effect on weight, food intake, blood and serum biochemical indicators in rats, there are pathological textures in the gastrointestinal tissue caused by diarrhea. Therefore, the safety threshold of Moringa homogenate should be ≤ 3 g/kg BW.

Protective effect of cinnamon extract against cobalt-induced multiple organ damage in rats

BackgroundThe role of oxidative stress and inflammation in cobalt (Co) toxicity has been the focus of previous studies. Cinnamon and its main components have been reported to have protective effects in various tissues with antioxidant and anti-inflammatory effects.AimsIn this study, the protective effect of cinnamon extract (CE) against possible Co-induced heart, kidney, and liver damage in rats was investigated biochemically.MethodsEighteen albino Wistar-type male rats were categorized into three groups (n = 6 per group): control (CG), CoCL2-administered (CoCL2), and CE + CoCL2-administered (CE + Co) groups. The CE + CoCL2 group was administered CE (100 mg/kg), and the CoCL2 and CG groups were administered distilled water orally by gavage. One hour after the administration, Co (150 mg/kg) was administered orally to the CE + CoCL2 and CoCL2 groups. This procedure was repeated once daily for 7 days. Then, biochemical markers were studied in the excised heart, kidney, and liver tissues.ResultsCoCL2 increased oxidants and proinflammatory cytokines and decreased antioxidants in heart, kidney, and liver tissues. Heart, kidney, and liver tissue were affected by Co damage. CE treatment suppressed the CoCL2-induced increase in oxidants and proinflammatory cytokines and decrease in antioxidants in heart, kidney, and liver tissues. CE treatment has been shown to attenuate cardiac damage by reducing serum troponin I (TpI) and creatine kinase-MB (CK-MB), renal damage by reducing creatinine and blood urea nitrogen (BUN), and liver damage by reducing alanine aminotransferase (ALT) and aspartate aminotransferase (AST).ConclusionCo induced the production of oxidants and proinflammatory parameters and antioxidant depletion in heart, kidney, and liver tissues of rats. Our experimental results show that CE protects heart, kidney, and liver tissues against oxidative and inflammatory changes induced by CoCLl2.

N1F(Improved-Nephropathy 1 Formula) Ameliorates Renal Interstitial Fibrosis via Inhibiting Extracellular Matrix Deposition and Regulating the FGF23/P38MAPK/Wnt Pathway.

Renal fibrosis is the primary pathway in the progression of chronic kidney disease (CKD) towards end-stage renal failure. The currently used drugs currently are ineffective, and their mechanisms of action remain unclear. This study aims to investigate the nephroprotective effect of Improved-Nephropathy 1 Formula (N1F) in a rat model of unilateral ureteral obstruction (UUO) and explore the potential mechanisms of N1F-containing serum in treating TGF-ß1-induced human renal tubular epithelial cells (HK-2). SD rats received 2-week continuous N1F gavage starting on day 2 after UUO. HK-2 cells were pretreated with a P38MAPK inhibitor for 1 h in vitro, followed by induction of the cells with TGF-ß1 and treatment with N1F 48 h later. The chemical composition of N1F was analyzed using high-performance liquid chromatography-Q-Orbitrap high-resolution liquid mass spectrometry. Renal function was assessed by measuring serum creatinine (Scr), blood urea nitrogen (BUN) and urine protein (Upro) levels. Hematoxylin and eosin (HE) and Masson's trichrome (Masson) staining were used to evaluate the extent of renal tissue damage and fibrosis. Western blotting, immunohistochemistry, and immunofluorescence were used to detect the protein levels of relevant indices. The RNA levels of the relevant indices were detected using real-time fluorescence quantitative PCR (RT-qPCR). We identified 361 chemical components in the water extract of N1F. These chemical components of N1F significantly reduced the area associated with interstitial fibrosis in the kidneys of UUO rats and the levels of serum creatinine, urea nitrogen, and urinary protein. Additionally, N1F decreased the protein levels of FGF23, Wnt1, ß-catenin and p-P38MAPK/P38MAPK, along with the expression of renalfibrosis-associated proteins, α-SMA, FN, Collagen III, and Vimentin in the renal tissues of the UUO rats, while enhancing klotho and DKK1 protein levels. In vitro experiments revealed that inhibition of P38MAPK signaling significantly suppressed the expression of proteins related to the Wnt signaling pathway, with a concomitant decrease in the expression of FGF23 and an increase in the expression of Klotho. Notably, the P38MAPK inhibitor (SB203580) had similar effects to N1F in altering the above-mentioned indices in vitro. N1F may exhibit potential therapeutic efficacy against renal fibrosis by inhibiting the FGF23/P38MAPK/Wnt signaling pathway, consequently inhibiting extracellular matrix deposition due to renal injury.

Exploring the metabolic implications of blue light exposure during daytime in rats

Excessive exposure to light is a global issue. Artificial light pollution has been shown to disrupt the body's natural circadian rhythm. To investigate the impacts of light on metabolism, we studied Sprague-Dawley rats chronically exposed to red or blue light during daytime or nighttime. Rats in the experimental group were exposed to extended light for 4 hours during daytime or nighttime to simulate the effects of excessive light usage. Strikingly, we found systemic metabolic alterations only induced by blue light during daytime. Furthermore, we conducted metabolomic analyses of the cerebrospinal fluid, serum, heart, liver, spleen, adrenal, cerebellum, pituitary, prostate, spermatophore, hypothalamus and kidney from rats in the control and blue light exposure during daytime. Significant changes in metabolites have been observed in cerebrospinal fluid, serum, hypothalamus and kidney of rats exposed to blue light during daytime. Metabolic alterations observed in rats encompassing pyruvate metabolism, glutathione metabolism homocysteine degradation, phosphatidylethanolamine biosynthesis, and phospholipid biosynthesis, exhibit analogous patterns to those inherent in specific physiological processes, notably neurodevelopment, cellular injury, oxidative stress, and autophagic pathways. Our study provides insights into tissue-specific metabolic changes in rats exposed to blue light during the daytime and may help explain potential mechanisms of photopathogenesis.

A Comprehensive Study to Determine the Residual Elimination Pattern of Major Metabolites of Amoxicillin–Sulbactam Hybrid Molecules in Rats by UPLC–MS/MS

Amoxicillin and sulbactam are widely used in animal food compounding. Amoxicillin–sulbactam hybrid molecules are bicester compounds made by linking amoxicillin and sulbactam with methylene groups and have good application prospects. However, the residual elimination pattern of these hybrid molecules in animals needs to be explored. In the present study, the amoxicillin–sulbactam hybrid molecule (AS group) and a mixture of amoxicillin and sulbactam (mixture group) were administered to rats by gavage, and the levels of the major metabolites of amoxicillin, amoxicilloic acid, amoxicillin diketopiperazine, and sulbactam were determined by UPLC–MS/MS. The residue elimination patterns of the major metabolites in the liver, kidney, urine, and feces of rats in the AS group and the mixture group were compared. The results showed that the total amount of amoxicillin, amoxicilloic acid, amoxicillin diketopiperazine, and the highest concentration of sulbactam in the liver and kidney samples of the AS group and the mixture group appeared at 1 h after drug withdrawal. Between 1 h and 12 h post discontinuation, the total amount of amoxicillin, amoxicilloic acid, and amoxicillin diketopiperazine in the two tissues decreased rapidly, and the elimination half-life of the AS group was significantly higher than that in the mixture group (p < 0.05); the residual amount of sulbactam also decreased rapidly, and the elimination half-life was not significantly different (p > 0.05). In 72 h urine samples, the total excretion rates were 60.61 ± 2.13% and 62.62 ± 1.73% in the AS group and mixture group, respectively. The total excretion rates of fecal samples (at 72 h) for the AS group and mixture group were 9.54 ± 0.26% and 10.60 ± 0.24%, respectively. These results showed that the total quantity of amoxicillin, amoxicilloic acid, and amoxicillin diketopiperazine was eliminated more slowly in the liver and kidney of the AS group than those of the mixture group and that the excretion rate through urine and feces was essentially the same for both groups. The residual elimination pattern of the hybrid molecule in rats determined in this study provides a theoretical basis for the in-depth development and application of hybrid molecules, as well as guidelines for the development of similar drugs.

EVALUATION OF ACUTE AND SUB-ACUTE TOXICITY OF METHANOLIC LEAF EXTRACT OF BAMBUSA VULGARIS ON MALE WISTAR RATS

Objectives: The study aims to evaluate the potential toxicity of methanolic extract of leaves of Bambusa vulgaris in male Wistar rats. Methods: The methanolic extract was prepared for acute and sub-acute toxicity to determine the lethal dose. During the acute toxicity study, male rats were orally administered with leaf extracts at single doses of 100, 500,800, and 2000 mg/kg according to OECD Guidelines 425 for 14 days. Sub-acute toxicity dose at 200, 400, and 800 mg/kg was studied by daily dosing on male rats for 30 days. Results: The acute toxicity study revealed no lethal effects and behavioral signs of toxicity at the tested doses indicating that LD50 is <2000 mg/kg. In a sub-acute study, non-significant changes in the body weight (p<0.05), body weight gain percentage (p<0.05) food intake (p<0.05), and relative organ weight percentage (p<0.05), and hematological parameters (p<0.05) of the rats were observed. A significant (p<0.05) increase in ALT level was observed dose at 800 mg/kg. Histopathological analysis showed mild cell distortion in the liver and kidney in male rats treated at 800 mg/kg of extract. Conclusion: Our study suggests that oral administration of methanolic leaf extract Bambusa vulgais is safe. However, a dose of 800 mg/kg may be toxic for the liver and kidney.

Transcriptomics and UHPLC-QQQ-MS analyses reveal the dysregulation of branched chain amino acids metabolism in renal fibrotic rats

The dysregulated levels of branched chain amino acids (BCAA) contribute to renal fibrosis in chronic kidney disease (CKD), yet specific analysis of BCAA contents and how they are regulated still remain unclear. It is therefore of great scientific interest to understand BCAA catabolism in CKD and develop a sensitive method for simultaneous determination of individual BCAA and their metabolites branched chain α-ketoacids (BCKA). In this work, the important role of BCAA metabolism that drives renal fibrosis in the process of CKD was first revealed by using transcriptomics. The key target genes controlling BCAA metabolism were then validated, that is, mRNA levels of BCKDHA and BCKDHB, the regulating rate-limiting enzymes during BCAA metabolism were abnormally reduced by quantitative PCR (qPCR), and a similar drop-off trend of protein expression of BCKDH, HIBCH and MCCC2 that are closely related to BCAA metabolism was also confirmed by western blotting. Furthermore, we established a novel strategy that simultaneously determines 6 individual BCAA and BCKA in serum and tissue. The method based on dansylhydrazine derivatization and ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-QQQ-MS) achieved to simultaneously determine the contents of BCAA and BCKA, which is efficient and stable. Compared with normal rats, levels of BCAA including leucine, isoleucine and valine in serum and kidney of CKD rats was decreased, while BCKA including α-ketoisocaproic acid, α-ketomethylvaleric acid and α-ketoisovaleric acid was increased. Together, these findings revealed the abnormality of BCAA metabolism in driving the course of kidney fibrosis and CKD. Our current study sheds new light on changes in BCAA metabolism during CKD, and may facilitate development of drugs to treat CKD and renal fibrosis.

The Possible Side Effects of Ziziphus spina-christi Extract on the Liver, Kidneys of Female Rats

Background: It has become important to provide advice and education to herbal remedies users to increase evidence-based awareness of their potential side effects. This research explores the side effects of the Ziziphus spina-christi (ZSC) aqueous extract on biochemical indicators and histopathological changes of the liver, kidneys of female rats. Methods: Fifteen female rats (five in each group) were orally administrated with the extract of ZSC at dosages of 10, 50 and 100 mg/kg bw/day for 14 days. Five other females were considered a control group, given only water. Clinical observations of toxicity were recorded during fifteen days. The biochemical biomarkers of liver (ALT and ALP enzymes) and kidneys (creatinine, uric acid and blood urea nitrogen) were evaluated in blood serum. Result: In addition, the histopathological alterations in organs were evaluated. The results did not show any clear toxicity on the animal’s weight or body organs. The alkaline phosphatase was elevated in female rats who received 50 mg and 100 mg of ZSC aqueous extract in comparison with the control group. The results did not show any significant changes in the serum glucose, BUN, creatinine, uric acid, alanine aminotransferase levels in female rats who received ZSC extract for 14 days. In conclusion, the results showed indications of side effects of the ZSC extract on liver enzymes after treating female rats with ZSC extract for 14 days.

A comparative study on the protective effects of cuminaldehyde, thymoquinone, and gallic acid against carbon tetrachloride-induced pulmonary and renal toxicity in rats by affecting ROS and NF-κB signaling

CCl4 toxicity is a fatal condition that can cause numerous organ dysfunctions. We evaluated and compared the protective effects of cuminaldehyde (CuA), thymoquinone (TQ), and gallic acid (GA) on CCl4-induced pulmonary and renal toxicity in rats. The impacts of these compounds on CCl4-induced oxidative stress, inflammation, and morphological alterations were examined. The results showed that the compounds under investigation prevented CCl4 from significantly increasing pulmonary and renal lipid peroxidation and NO levels, as well as massively depleting GSH levels and GPX and SOD activities. Moreover, they suppressed the CCl4-induced increase in mucus secretion in the lung and upregulated the gene expression of pulmonary and renal NF-ҡB, iNOS, TNF-α, and COX-2. The heatmap cluster plots showed that GA and TQ had better protective potencies than CuA. The external organ morphology, histopathological results, and chest X-ray analysis confirmed the toxicity of CCl4 and the protective influences of the tested compounds in both the lungs and kidneys of rats. These compounds displayed predicted competitive inhibitory effects on iNOS activity and may block the IL-13α2 receptor, as revealed by molecular docking analysis. Thus, CuA, TQ, and GA, particularly the latter two, are prospective protective compounds against the pulmonary and renal toxicity caused by CCl4.

Effects of ethanolic extract of <i>Aristolochia repens</i> on biochemical and histological changes in high-fat diet and streptozotocin-induced rats

The study evaluated the lipid profile, electrolytes and histological changes in diabetic Wistar rats treated with stem bark extract of Aristolochia repens. Type 2 diabetes was induced in rats with high-fat diet and low dose of streptozotocin (35 mg/kg body weight). A total of thirty-five Wistar rats were fed with high-fat diet for eight (8) weeks, while five (5) Wistar rats (Sham control) were fed with low-fat diet. Non-induced male Wistar rats were treated with distilled water (Sham control); while the induced male Wistar rats were treated with distilled water, metformin (100 mg/kg), 50, 100 and 200 mg/kg ethanolic extract of Aristolonchia repens stem bark for 28 days. The rats were sacrificed; liver, kidney and serum were collected and analyzed for serum glucose, lipid profile, electrolytes and histological changes. Treatment with ethanolic extract of Aristolonchia repens stem bark significantly reduced the body weight, glucose level, total cholesterol concentration, triglycerides, low-density lipoprotein concentration and atherogenic indices. Mild pathological alterations were observed in the liver and kidney of diabetic rats at dosage above 50 mg/kg. However, the extract reduced the blood glucose, lipids and improve the selected electrolyte balance. This study suggests that ethanolic extract of stem bark of A. repens possesses antihyperglycaemia and antilipidaemic potential and might not be safe at dosage above 50 mg/kg.

Eplerenone reduces lymphangiogenesis in the contralateral kidneys of UUO rats.

Inflammation and fibrosis often occur in the kidney after acute injury, resulting in chronic kidney disease and consequent renal failure. Recent studies have indicated that lymphangiogenesis can drive renal inflammation and fibrosis in injured kidneys. However, whether and how this pathogenesis affects the contralateral kidney remain largely unknown. In our study, we uncovered a mechanism by which the contralateral kidney responded to injury. We found that the activation of mineralocorticoid receptors and the increase in vascular endothelial growth factor C in the contralateral kidney after unilateral ureteral obstruction could promote lymphangiogenesis. Furthermore, mineralocorticoid receptor activation in lymphatic endothelial cells resulted in the secretion of myofibroblast markers, thereby contributing to renal fibrosis. We observed that this process could be attenuated by administering the mineralocorticoid receptor blocker eplerenone, which, prevented the development of fibrotic injury in the contralateral kidneys of rats with unilateral ureteral obstruction. These findings offer valuable insights into the intricate mechanisms underlying kidney injury and may have implications for the development of therapeutic strategies to mitigate renal fibrosis in the context of kidney disease.

Dual-specificity phosphatase 26 protects against kidney injury caused by ischaemia-reperfusion through restraint of TAK1-JNK/p38-mediated apoptosis and inflammation of renal tubular epithelial cells

Dual-specificity phosphatase 26 (DUSP26) acts as a pivotal player in the transduction of signalling cascades with its dephosphorylating activity. Currently, DUSP26 attracts extensive attention due to its particular function in several pathological conditions. However, whether DUSP26 plays a role in kidney ischaemia-reperfusion (IR) injury is unknown. Aims of the current work were to explore the relevance of DUSP26 in kidney IR damage. DUSP26 levels were found to be decreased in renal tubular epithelial cells following hypoxia-reoxygenation (HR) and kidney samples subjected to IR treatments. DUSP26-overexpressed renal tubular epithelial cells exhibited protection against HR-caused apoptosis and inflammation, while DUSP26-depleted renal tubular epithelial cells were more sensitive to HR damage. Upregulation of DUSP26 in rat kidneys by infecting adenovirus expressing DUSP26 markedly ameliorated kidney injury caused by IR, while also effectively reducing apoptosis and inflammation. The mechanistic studies showed that the activation of transforming growth factor-β-activated kinase 1 (TAK1)-JNK/p38 MAPK, contributing to kidney injury under HR or IR conditions, was restrained by increasing DUSP26 expression. Pharmacological restraint of TAK1 markedly diminished DUSP26-depletion-exacebated effects on JNK/p38 activation and HR injury of renal tubular cells. The work reported a renal-protective function of DUSP26, which protects against IR-related kidney damage via the intervention effects on the TAK1-JNK/p38 axis. The findings laid a foundation for understanding the molecular pathogenesis of kidney IR injury and provide a prospective target for treating this condition.

Time Efficient Ultrasound Localization Microscopy Based on A Novel Radial Basis Function 2D Interpolation.

Ultrasound localization microscopy (ULM) allows for the generation of super-resolved (SR) images of the vasculature by precisely localizing intravenously injected microbubbles. Although SR images may be useful for diagnosing and treating patients, their use in the clinical context is limited by the need for prolonged acquisition times and high frame rates. The primary goal of our study is to relax the requirement of high frame rates to obtain SR images. To this end, we propose a new time-efficient ULM (TEULM) pipeline built on a cutting-edge interpolation method. More specifically, we suggest employing Radial Basis Functions (RBFs) as interpolators to estimate the missing values in the 2-dimensional (2D) spatio-temporal structures. To evaluate this strategy, we first mimic the data acquisition at a reduced frame rate by applying a down-sampling (DS = 2, 4, 8, and 10) factor to high frame rate ULM data. Then, we up-sample the data to the original frame rate using the suggested interpolation to reconstruct the missing frames. Finally, using both the original high frame rate data and the interpolated one, we reconstruct SR images using the ULM framework steps. We evaluate the proposed TEULM using four in vivo datasets, a Rat brain (dataset A), a Rat kidney (dataset B), a Rat tumor (dataset C) and a Rat brain bolus (dataset D), interpolating at the in-phase and quadrature (IQ) level. Results demonstrate the effectiveness of TEULM in recovering vascular structures, even at a DS rate of 10 (corresponding to a frame rate of sub-100Hz). In conclusion, the proposed technique is successful in reconstructing accurate SR images while requiring frame rates of one order of magnitude lower than standard ULM.

Effect of Ethanolic Leaf Extract of Piper guineense (Uziza) on the Kidneys of Paraquat-Induced Adult Male Wistar Rats

Effect of Ethanolic Leaf Extract of Piper guineense (Uziza) on the Kidneys of Paraquat-Induced Adult Male Wistar Rats

Reversible assembly and disassembly of V-ATPase during the lysosome regeneration cycle.

Regulation of the luminal pH of late endocytic compartments in continuously fed mammalian cells is poorly understood. Using normal rat kidney fibroblasts, we investigated the reversible assembly/disassembly of the proton pumping V-ATPase when endolysosomes are formed by kissing and fusion of late endosomes with lysosomes and during the subsequent reformation of lysosomes. We took advantage of previous work showing that sucrosomes formed by the uptake of sucrose are swollen endolysosomes from which lysosomes are reformed after uptake of invertase. Using confocal microscopy and subcellular fractionation of NRK cells stably expressing fluorescently tagged proteins, we found net recruitment of the V1 subcomplex during sucrosome formation and loss during lysosome reformation, with a similar time course to RAB7a loss. Addition of invertase did not alter mTORC1 signalling, suggesting that the regulation of reversible V-ATPase assembly/disassembly in continuously fed cells differs from that in cells subject to amino acid depletion/refeeding. Using live cell microscopy, we demonstrated recruitment of a fluorescently tagged V1 subunit during endolysosome formation and a dynamic equilibrium and rapid exchange between the cytosolic and membrane bound pools of this subunit. We conclude that reversible V-ATPase assembly/disassembly plays a key role in regulating endolysosomal/lysosomal pH in continuously fed cells.

Antioxidant Vitamins (Vit C and E) Protect the Kidney from Glyphosate- Exposed Renal Damage in Experimental Rats - A Molecular Study

Glyphosate is a widely used herbicide and the active ingredient in Monsanto's Roundup, among other herbicide formulations. Vitamin C (ascorbic acid) and vitamin E (tocopherol) are both antioxidants that play important roles in reducing oxidative DNA damage and protecting cells from the harmful effects of free radicals. The study aimed to analyse the effect of antioxidant vitamins (vitamins C and E) on the expression of IGF-1 and KIM-1 molecules in the kidneys of glyphosate-exposed rats. Adult male Albino Wistar rats were grouped into 3. Group I: Control; Group II: Glyphosate treated (100mg); Group III: Glyphosate+ vitamin E & C treated. Serum, insulin, and mRNA expression of insulin-like growth factor (IGF-1) and kidney injury molecules (KIM-1) mRNA expression by RT-PCR. Serum insulin was measured by the ELISA method. Glyphosate-exposed rats developed hyperglycaemia and hyper compared to control but antioxidant vitamins (Vitamin C and E) supplementation, reduced the glyphosate mediate increased fasting blood glucose and insulin near to that of the control level. Similarly, mRNA expression of both IGF-1 and KIM-1 were also significantly altered due to glyphosate induction whose effects were found to be reduced when vitamins C and E were treated. It is concluded that glyphosate exposure causes renal damage and develops diabetic nephropathy by modulating the expression of IGF-1 and KIM-1 molecules in the kidney. Vitamin C and E potentially were able to reduce the detrimental changes caused by glyphosate. Hence, vitamin C and E supplements could be considered therapeutic drugs for diabetic nephropathy.

Comparison of the effects of gasoline burn and chromic acid burn on different internal organs and immune functions in rats.

Burn as physical injury ranks as the fourth most prevalent trauma across the world. In this study, we aimed to compare the impact of gasoline burn and chromic acid burn on the internal organs and immune functions in rats. The results showed that the levels of methemoglobin (MHb) to total hemoglobin (Hb) as well as the Cr6+ content showed significant elevation in the chromic acid burn group relative to the gasoline burn group. HE staining was used to evaluate the histological changes in the injured tissues as well as the tissues excised from internal organs. We found that chromic acid burn-induced more severe damage to rat tissues. Gasoline burn showed no significant impact on the intestinal tissues of rats, while the chromic acid burn-induced increased cell death in rat intestines. Moreover, the results of HE staining also revealed that gasoline burn and chromic acid burn showed no evident impact on rat hearts. Gasoline burn also showed no significant effects on the liver, lungs and kidneys of rats, while the chromic acid burn caused injuries to such internal organs in comparison with the control and gasoline burn groups. In addition, the MPO activity was higher in the liver, intestine, lungs and kidneys of rats with chromic acid burn. Furthermore, the expression of inflammation response cytokines was examined in the serum of rats. The results demonstrated that the levels of IL-6, IL-1β and TNF-α showed a significant increase in both the gasoline burn and chromic acid burn groups of rats relative to the control, and the levels were higher in the chromic acid burn group in comparison with the gasoline burn group. In conclusion, the chromic acid burn-induced more severe organ injury, inflammation and immune response compared with the gasoline burn, which may provide reference data for the clinical treatment of patients with different burn injuries.