Rat Renal Tissue Research Articles

Phytochemical investigation and nephroprotective potential of Sida cordata in rat

BackgroundPlants are an efficient source of natural antioxidant against free radicals causing kidney damages. Sida cordata ethyl acetate fraction has been reported for strong in vitro antioxidant potency, previously. In the present study, our objective was to evaluate its in vivo antioxidant potency against CCl4 induced nephrotoxicity and investigates the bioactive phytochemicals by HPLC-DAD analysis.MethodsPhytochemical analysis was performed by HPLC-DAD methodology. For in vivo study, 42 male Sprague-Dawley rats were treated with alternatively managed doses for 60 days. Group I animals were remained untreated. Group II animals were treated with vehicle (1 mL of olive oil) by intragastric route on alternate days. Group III was treated with 30% CCl4 (1 mL/kg b.w.) i.p. Group IV was treated with 30% CCl4 (1 mL/kg b.w.) i.p and silymarin intragastric. Group V and VI rats were treated with 30% CCl4 and SCEE (150 and 300 mg/kg b.w., respectively) intragastric. Group VII animals were treated with SCEE (300 mg/kg b.w.) intragastrically. Blood parameters, Serum proteins and urine profile were investigated. Activities of tissue enzyme i.e. catalase, peroxidase, superoxide dismutase, glutathione-S-transferase, glutathione reductase, GSH and γ-GT were evaluated. Histopathological observations, total protein contents, lipid peroxidation, DNA damage and relative weight were also analyzed.ResultsGallic acid, catechin and caffeic acid were identified in SCEE fraction by HPLC-DAD. Decrease in the count of red blood cells, neutrophils, eosinophils and concentration of hemoglobin whereas increase in lymphocyte count and estimation of sedimentation rate (ESR) with 1 mL CCl4 (30% in Olive oil) administration (30 doses in 60 days) was restored dose dependently with co-treatment of SCEE (150 and 300 mg/kg b.w.). Treatment of rats with CCl4 markedly (P < 0.01) increased the count of urinary red blood cells and leucocytes, concentration of urea, creatinine and urobilinogen and specific gravity whereas creatinine clearance was reduced. Serum level of total protein, albumin, globulin, nitrite, creatinine and blood urea nitrogen (BUN) was significantly increased (P < 0.01) by CCl4 treatment. The activity of antioxidant enzymes; catalase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and glutathione reductase and content of reduced glutathione was decreased (P < 0.01) significantly. However, increased concentration (P < 0.01) of thiobarbituric acid reactive substances and histopathological injuries were noticed in the renal tissues of rats after the treatment with CCl4. Co-administration of SCEE, dose dependently, protected the alterations in the studied parameters of rats at 150 and 300 mg/kg b.w. The present study revealed that SCEE could be used as a possible remedy for renal toxicity abnormalities.ConclusionThese results are an evidence of the renal protective role of S.cordat ethyl acetate fraction against CCl4 induced nephrotoxicity in rats which may be due to its antioxidant compounds.

Panax notoginseng saponins mitigate cisplatin induced nephrotoxicity by inducing mitophagy via HIF-1α.

We investigated the role of HIF-1α in the mitigation of cisplatin-induced nephrotoxicity by Panax notoginseng saponins (PNS) in a rat model. Serum creatinine (Scr), blood urea nitrogen (BUN) and urinary N-acetyl-β-D-glucosaminidase (NAG) levels were all elevated in cisplatin treated rats. PNS reduced Scr, BUN and NAG levels in the presence or absence of the HIF-1α inhibitor 2-methoxyestradiol (2ME2). PNS also reduced the high tubular injury scores, which corresponded to renal tubular damage in cisplatin-treated rats and which were exacerbated by 2ME2. Renal tissues from PNS-treated rats showed increased HIF-1α mRNA and nuclear localized HIF-1α protein. Moreover, PNS treatment increased BNIP3 mRNA as well as LC3-II, BNIP3 and Beclin-1 proteins and the LC3-II/LC3-I ratio in rat renal tissues. This suggested that PNS treatment enhanced HIF-1α, which in turn increased autophagy. This was confirmed in transmission electron micrographs of renal tissues that showed autophagosomes in PNS-treated renal tissues. These findings demonstrate that PNS mitigates cisplatin-induced nephrotoxicity by enhancing mitophagy via a HIF-1α/BNIP3/Beclin-1 signaling pathway.

Diabetes increases interleukin-17 levels in periapical, hepatic, and renal tissues in rats

ObjectivesThis study aimed to evaluate the association between endodontic infection and diabetes on interleukin-17 levels in periapical, hepatic, and renal tissues of rats. DesignForty male rats were divided into groups: normoglycemic rats (N), normoglycemic rats with apical periodontitis (N-AP), rats with experimental diabetes (ED), and rats with experimental diabetes and apical periodontitis (ED-AP). Diabetes was induced by intravenous streptozotocin injection, and blood sugar levels were monitored to confirm disease development. Apical periodontitis (AP) was induced by pulp exposure to the oral environment during 30days. After 30days, hepatic and renal tissues were obtained, and IL-17 levels were quantified by ELISA. The right hemi-jaw was used to quantify IL-17 levels by immunohistochemistry. The values obtained in parametric tests were tabulated and analyzed statistically by analysis of variance (ANOVA) and Tukey tests, and the values obtained for scores were statistically analyzed by using the Kruskal-Wallis and Dun tests. The level of significance was set at 5%. ResultsED and ED-AP groups expressed significantly higher IL-17 levels in both hepatic and renal tissues (p<0.05), compared to N and N-AP groups. Apical periodontitis (AP) in ED-AP group was significantly more severe than that in N-AP group (p<0.05). Furthermore, there was a significantly larger increase in the IL-17 levels in ED-AP group compared to N group (p<0.05). ConclusionOur results indicate that diabetes increases IL-17 levels in hepatic and renal tissues and also enhances IL-17 production in apical periodontitis area of rats.

Histoenzymic determination of activity of alkaline phosphatase and succinate dehydrogenase in the renal tissue of rats at the early stage of formation of experimental diabetes mellitus

Histoenzymic determination of activity of alkaline phosphatase and succinate dehydrogenase in the renal tissue of rats at the early stage of formation of experimental diabetes mellitus

Protective effect of Sargussum vulgare sulfated polysaccharide against molecular, biochemical and histopathological damage caused by alloxan in experimental diabetic rats

Diabetes is a serious condition that is linked to the development of oxidative stress. In the context of enhancing the biodiversity of Tunisia's flora, this study aimed to evaluate the effect of Sargussum vulgare sulfated polysaccharide (SVSP) on hyperglycemia and diabetes complications in the alloxan-induced diabetic rats.Our results showed a disturbance of carbohydrate, lipid, hematological and histopathological parameters, an increase in the α-amylase enzyme activity and damage to the pancreatic, hepatic and renal tissues in rats rendered diabetic by alloxan. In contrast, treatment with SVSP resulted in a correction of fasting and postprandial blood glucose and HbA1c through inhibition of pancreatic α-amylase. We also noticed an improvement in hemogram parameters and an attenuation of the pancreatic oxidative stress markers as well as histological protections. On the other hand, the administration of SVSP to diabetic rats caused, besides the correction of glycemic and lipid parameters, a good defense against hepatotoxicity and nephrotoxicity of diabetes as well as good antioxidant status and histological protections.

Effects of early overnutrition on the renal response to Ang II and expression of RAAS components in rat renal tissue

Background and aimsThe aim of this study was to analyze the effects of early overnutrition (EON) on the expression of the renin angiotensin aldosterone system (RAAS) components in renal cortex, renal arteries and renal perivascular adipose tissue (PVAT), as well as the vascular response of renal arteries to Angiotensin II (Ang II). Methods and resultsOn birth day litters were adjusted to twelve (L12-control) or three (L3-overfed) pups per mother. Half of the animals were sacrificed at weaning (21 days old) and the other half at 5 months of age. Ang II-induced vasoconstriction of renal artery segments increased in young overfed rats and decreased in adult overfed rats. EON decreased the gene expression of angiotensinogen (Agt), Ang II receptors AT1 and AT2 and eNOS in renal arteries of young rats, while it increased the mRNA levels of AT-2 and ET-1 in adult rats. In renal PVAT EON up-regulated the gene expression of COX-2 and TNF-α in young rats and the mRNA levels of renin receptor both in young and in adult rats. On the contrary, Ang II receptors mRNA levels were downregulated at both ages. Renal cortex of overfed rats showed increased gene expression of Agt in adult rats and of AT1 in young rats. However the mRNA levels of AT1 were decreased in the renal cortex of overfed adult rats. ConclusionEON is associated with alterations in the vascular response of renal arteries to Ang II and changes in the gene expression of RAAS components in renal tissue.

RETRACTED: Diallyl trisulfide, a garlic polysulfide protects against As-induced renal oxidative nephrotoxicity, apoptosis and inflammation in rats by activating the Nrf2/ARE signaling pathway

Arsenic (As) contamination is an extremely dangerous global environmental problem as it can enter into the food chain and become bio-accumulated, endangering human health. Chronic As intoxication leads to undesirable toxic effects in various organ systems of the body, especially the kidney. Diallyl trisulfide (DATS) is an organosulfur compound which has been widely known for its uses as antibacterial, antitumorogenic, antioxidant agent and has been also reported to have anti-apoptotic and anti-inflammatory properties. In the present work, we intend to investigate the protective role of DATS, a garlic organosulfur compound in preventing the As-induced oxidative stress mediated renal injury in rats. Study design The activity of DATS to antagonize As-induced renal oxidative toxicity was analyzed using rats as an in vivo model. We investigated the nephroprotective effect of DATS on As treated rats by performing various serological, biochemical, molecular and histological studies. The activation of Nrf2 was investigated using western blot. The data showed that As exposure significantly increased the serum and urine nephritic, oxidative stress, apoptosis and inflammatory markers in the renal tissue of rats. As intoxication also decreased the antioxidant status of the renal tissue along with the disturbances in the membrane bound ATPases. As nephrotoxicity was further confirmed with the altered morphological and ultrastructural changes in the renal tissue. Conversely, the DATS pre-administration effectively recuperate the altered renal variables by As, which has been further supported by the histological and ultrastructural observations. This counteraction was achieved partially via the activation of Nrf2-ARE pathway through the activation of Akt. These findings explicate the prospective use of DATS as a promising organosulfur compound against As-induced renal oxidative dysfunction in rats.

Rotenone induces nephrotoxicity in rats: oxidative damage and apoptosis

Previous studies have examined rotenone toxicity on the human central nervous system, especially in the pathogenesis of Parkinson’s disease, but few have investigated the effects of rotenone on the kidney. Here, rotenone-induced nephrotoxicity was evaluated by determining morphological, biochemical, oxidative stress-related, and apoptotic factor alterations in rat renal tissue. Morphological and biochemical analyzes showed that rotenone administration to rats damaged renal tissue. Western blot results revealed that rotenone-induced oxidative damage, causing overproduction of glutathione, malonaldehyde, and reactive oxygen species (ROS), and inhibiting superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity. Rotenone also decreased the mitochondrial membrane potential and increased voltage-dependent anion channel (VDAC), caspase-3, and caspase-9 protein levels, indicating an association of apoptosis with renal damage. Our results suggest that glutathione, malonaldehyde, and ROS may be signals of rotenone-induced oxidative damage, and that the mitochondrial pathway plays a key role in apoptosis of renal cells following rotenone administration.

Implication of hyperoxaluria on osteopontin and ER stress mediated apoptosis in renal tissue of rats

Hyperoxaluria is a stress that leads to calcium oxalate crystal deposition which further causes inflammation and renal cell necroptosis. Many studies have linked osteopontin expression with apoptosis and inflammation but so far its association with apoptosis with regard to hyperoxaluria is undiscovered. Moreover, a recent report has suggested that osteopontin induces endoplasmic reticulum stress and subsequently apoptosis in myocytes. In this study, the impact of hyperoxaluria on the modulation of osteopontin expression and endoplasmic reticulum (ER) stress mediated apoptosis in rats is explored. Hyperoxaluria was induced in rats by three different doses viz. ethylene glycol alone, ethylene glycol and ammonium chloride together and third group were fed with hydroxyl-l-proline. After hyperoxaluria induction rats were sacrificed and renal tissue was analysed for crystal depositions, osteopontin expression, inflammation, ER stress and subsequent unfolded protein response intermediates (UPR). Altered histoarchitecture of renal tissue and elevated levels of reactive oxygen species (ROS) along with the presence of calcium oxalate crystals were observed in the hyperoxaluric groups. As expected, inflammation and apoptosis was significantly high in all hyperoxaluria groups. Osteopontin expression showed significant up-regulation following hyperoxaluria. Further, a similar trend between expression of osteopontin and elevated ER stress level was observed. Moreover, UPR intermediates expression was also concurrent with osteopontin levels. It is observed that the extent of calcium oxalate crystal deposition is directly associated with the expression of osteopontin, inflammation and ER stress. The results advocate possible association of osteopontin with ER stress, thus suggesting that the ER could be a new target for developing therapeutic regimes for kidney stones.

EXPRESSION miRNA-21 IN RENAL TISSUE AND URINE IN RATS WITH UNILATERAL URETERAL OBSTUCTION

THE AIM: to estimate the level of expression miRNA-21 inthe urine and renal tissue in rats with unilateral ureteral obstruction (UUO). MATHERIAL AND METHODS. UUO was induced by ligation of the left ureter in male Wistar rats (n=10). Follow-up period was 14 days after UUO modeling. Urine was collected one day before the operation (UmiRNA21C), and one day before the end of experiment (UmiRNA21I ) during 24 hours. Before releasing animal out of experiment collected urine from left kidney pelvis (UmiRNA21O) and tissue of left kidney (KmiRNA21O) and right kidney (KmiRNA21I). MiRNA-21 expression in kidney tissues and urine was carried out with reaction amplification (RealTime PCR-protocol). Calculation was realized by 2-deltaCtmethod. Statistical analysis was performed with Wilcoxon test and Spearman correlation coefficient. Results are demonstrated as median [low – upper quartile]. RESULTS. UmiRNA21I(3.78[2.0-5.28]) and UmiRNA21O(3.78[3.25-3.82]) were significantly higher than UmiRNA21C (1.15[0.71-1.74]; P=0.0125 and P=0.0069, respectively). UmiRNA21I and UmiRNA21O values were practically equal. In kidneys with UUO the tissue levels of miRNA21 expression was a higher than in contralateral organ (P=0,0926). Revealed direct correlation between KmiRNAIand KmiRNAO(RS=0,770, P=0,0092). CONCLUSION. UOO can cause specific changes in the expression, distribution and excretion of micro RNA-21 and its role in the development of renal tubulointestitsial fibrosis requires further studies. Key words: miRNA-21, tubulointerstitial fibrosis, unilateral ureteral obstruction

BMP-7 enhances SnoN mRNA expression in renal tubular epithelial cells under high-glucose conditions.

The present study aimed to identify any association between bone morphogenetic protein-7 (BMP-7) and the expression of the transcriptional co-repressor Ski-related novel protein N (SnoN), in addition to alterations in tubulointerstitial fibrosis, during the development and progression of diabetic nephropathy (DN). Streptozotocin was injected into the tail veins of 20 healthy and specific pathogen-free male Sprague-Dawley rats. The rats were sacrificed to detect the appropriate biochemical indicators of renal pathological alterations following 24 weeks. Then, various doses of human recombinant BMP-7 were added to high glucose-cultured NRK-52E cells. Immunohistochemistry, immunofluorescence staining and western blotting were used to determine the expression of SnoN, BMP-7, Smad ubiquitin regulatory factor (Smurf)2, Arkadia, E-cadherin, α-smooth muscle actin and Collagen III. Reverse transcription-quantitative polymerase chain reaction was used to detect SnoN mRNA expression. With the progression of DN, the expression of BMP-7 in rat renal tissue was downregulated, whereas the expression of Smurf2 and Arkadia increased. Furthermore, the expression of SnoN mRNA increased however the expression of SnoN protein decreased, accompanied by renal tubular epithelial cell mesenchymal transition, extracellular matrix (ECM) deposition and severe renal function disorder. The exogenous recombinant human BMP-7 alleviated high glucose-induced phenotypic transformation and ECM synthesis of NRK-52E in vitro and upregulated SnoN transcription and protein expression, however no effect was observed on the expression of Smurf2 and Arkadia. BMP-7 may ameliorate DN and renal fibrosis via increasing the expression of SnoN mRNA and protein in renal tubular epithelial cells, rather than directly inhibiting the degradation of SnoN by E3 ubiquitin ligase.

The changes of β-arrestin2 and microtubule-associated protein light chain 3 in renal tissue of rats with renal ischemia reperfusion injury

Objective To investigate the expression of β-arrestin2 and microtubule-associated protein light chain(LC)3 in renal of rat with acute renal ischemia reperfusion injury, and to analyze the relationship between them and renal injury. Methods Fifty-four male SD rat(3-4 weeks old) were randomly divided into three groups: control group, sham group, acute ischemic reperfusion injury group.We established the acute renal ischemia reperfusion injury model through removing the right kidney and clamping the left renal for 45 minutes with noninvasive arterial clip.We obtained the kidney and blood samples respectively at 12 h, 24 h, 36 h, 48 h, 72 h, 96 h after the surgery.Expressions of β-arrestin2 and LC3 protein were detected by the immunohistochemistry method and Western blot method.The renal function and morphological changes were assessed. Results Compared with control group and sham group, the serum creatinine and kidney pathological grading of acute ischemia reperfusion injury group obviously rised.The kidney injury was the most serious at the 24 h after acute ischemic reperfusion injury.The expressions of β-arrestin2 and LC3 were little in the control group and sham group.However, the expressions of these two indicators were obviously higher and reached the peak at the 12 h after acute ischemia reperfusion injury.All these results suggested that the changes of these two indicators were anterior to the histopathological changes.The expressions of β-arrestin 2 and LC3 protein were in positive correlation with the kidney injury(r=0.821, P<0.05; r=0.913, P<0.05). Conclusion In the acute renal ischemia-reperfusion injury, β-arrestin2 may be as a kind of upstream regulatory protein involving in the kidney pathological process through the regulation of the autophagy. Key words: β-arrestin2; Microtubule-associated protein light chain 3; Acute kidney injury; Ischemic reperfusion injury,

Protective Effect of Exacum lawii on Cisplatin-induced Oxidative Renal Damage in Rats.

Background:Exacum lawii (Gentianaceae), a bitter herb conventionally used in kidney diseases and eye problems, endemic to the Western coast and Southern part of India.Aim:Folklore reports encourage the author to explore the nephroprotective effect of the standardized ethanolic extract of E. lawii against cisplatin-induced renal toxicity in the rat to scientifically validate its traditional use.Materials and Methods:Ethanolic extract of the whole plant of E. lawii was standardized with swertiamarin (secoiridoid glycoside) using high-performance liquid chromatography and tested for subacute toxicity according to the OECD guidelines. Nephroprotective potential at different doses of extract was evaluated against cisplatin (6 mg/kg, intraperitoneal) in experimental rats. The changes in serum renal toxicity markers, renal tissue oxidative stress biomarkers, and proinflammatory cytokines level were measured. To estimate the change in oxidative status of renal tissues, DNA and single viable cells were isolated from treated rat kidney, DNA fragmentation assay and flow cytometric analysis of reactive oxygen species (ROS) were performed. Histopathology of renal tissues was also examined.Results:Swertiamerin was found to be 119.59 mg/g of extract. Administration of E. lawii extract (ELE) restored the biochemical parameters. It also decreases the elevated proinflammatory cytokines level in kidney tissues and protected rat kidneys from oxidative stress in rats. Nephroprotective activity was validated by estimating ROS production in kidney live cells and DNA damage in kidney tissue. The histological architecture was also conserved.Conclusion:ELE showed significant renal protection against cisplatin through reducing oxidative stress and inflammation.SUMMARY High-performance liquid chromatography standardisation of Exacum lawii extract with Swertiamerin and its subacute toxicity studyNephroprotective activity of Exacum lawii extract and Swertiamerin was evaluated in cisplatin-induced model and justified by various biochemical parameters and histopathological studyRole of oxidative stress in cisplatin-induced nephrotoxicity was confirmed by measuring levels of antioxidant markers and proinflammatory cytokines in rat renal tissues, ROS estimation by flow cytometry and DNA fragmentation assay by gel electrophoresis in renal cells. Abbreviations used: ELE: Exacum lawii ethanolic extract; WHO: World Health Organization; SOD: Superoxide dismutase; CAT: Catalase; MDA: Malondialdehyde; HPTLC: High performance thin layer chromatography; p.o.: Per oral; i.p.: Intraperitoneal; TNF-α: Tumor necrosis factor alpha; IL-1β: Interleukin 1 beta; IL-6: Interleukin 6; ROS: Reactive oxygen species.

Loss of the Protein Cystathionine β-Synthase During Kidney Injury Promotes Renal Tubulointerstitial Fibrosis

Background/Aims: Renal tubulointerstitial fibrosis (TIF) is the common pathway of progressive chronic kidney disease. Inflammation has been widely accepted as the major driving force of TIF. Cystathionine β-synthase (CBS) is the first and rate-limiting enzyme in the transsulfuration pathway. CBS is considered to play protective role in liver and pulmonary fibrosis, but its role in TIF remains unknown. The purpose of this study was to investigate the potential role and mechanism of CBS in renal inflammation and TIF. Methods: Renal function, tubulointerstitium damage index score, extracellular matrix (ECM) deposition, and the expressions of collagen I, collagen III, fibronectin, CD3, CD68, IL-1β, TNF-α were measured in sham operation and unilateral ureteral obstruction (UUO) rats. Proteomics and gene array analysis were performed to screen differentially expressed molecules in the development of renal inflammation and TIF in UUO rats. The expression of CBS was detected in patients with obstructive nephropathy and UUO rats. We confirmed the expression of CBS using western blot and real-time PCR in HK-2 cells. Overexpression plasmid and siRNA were transfected specifically to study the possible function of CBS in HK-2 cells. Results: Abundant expression of CBS, localized in renal tubular epithelial cells, was revealed in human and rat renal tissue, which correlated negatively with the progression of fibrotic disease. Expression of CBS was dramatically decreased in the obstructed kidney from UUO rats as compared with the sham group (SHM). In addition, knocking down CBS exacerbated extracellular matrix (ECM) deposition, whereas CBS overexpression attenuated TGF-β1-induced ECM deposition in vitro. Inflammatory and chemotactic factors were also increased in CBS knockdown HK-2 cells stimulated by IL-1β. Conclusions: These findings establish CBS as a novel inhibitor in renal fibrosis and as a new therapeutic target in patients with chronic kidney disease.

The Endocannabinoid, Anandamide, Induces Cannabinoid Receptor-Independent Cell Death in Renal Proximal Tubule Cells

Background: The endocannabinoid (EC) system is well characterized in the central nervous system but scarcely studied in peripheral organs. In this paper, we newly identify the effect of the EC anandamide (AEA) upon renal proximal tubule cells. Methods: Measurement of lactate dehydrogenase (LDH) release after treatment of primary renal proximal tubule cells (RPTEC) and renal carcinoma cell line (Caki-1) with AEA, arachidonic acid (AA), ethanolamide (EtAm), EC receptor CB1 antagonist (AM251), CB2 receptor antagonist (SR144528), TRPV1 receptor antagonist (capsazepine), degradation enzyme fatty acid amide hydrolase (FAAH) antagonist (URB597), antioxidants GSH-EE; Trolox, GSH depletor BSO, membrane cholesterol depletor (MCD), apoptosis inhibitor zVAD, necroptosis inhibitor Nec-1 or ferroptosis inhibitor Fer-1. Western blot and qRT-PCR analysis plus determination of reactive oxygen species (ROS) via H2-DCFDA were performed. Histology for EC enzymes, N-acetylphosphatidylethanolamine-hydrolyzing phospholipase D (NAPE-PLD) and FAAH, as well as the determination of physiological levels of ECs in human and rat renal tissue via liquid chromatography were conducted. Results: AEA both dose- and time-dependently induces cell death in RPTEC and Caki-1 within hours, characterized by cell blebbing, not influenced by blocking the described EC receptors by AM251, SR144528, capsazepine or FAAH by URB597 or MCD. Cell death is mediated via ROS. There is no difference found in the histology of the enzymes FAAH and NAPE-PLD in human renal tissue with interstitial nephritis. Blocking of apoptotic, necroptotic or ferroptotic cell death does not lead to a reduction in LDH release in vitro. Conclusion: The endocannabinoid anandamide induces cell death in renal proximal tubule cell in a time- and dose-dependent manner. This pathway is mediated via ROS and is independent of cannabinoid receptors, membrane cholesterol or FAAH activity.

Objective detection of apoptosis in rat renal tissue sections using light microscopy and free image analysis software with subsequent machine learning: Detection of apoptosis in renal tissue.

The current study proposes an automated machine learning approach for the quantification of cells in cell death pathways according to DNA fragmentation. A total of 17 images of kidney histological slide samples from male Wistar rats were used. The slides were photographed using an Axio Zeiss Vert.A1 microscope with a 40x objective lens coupled with an Axio Cam MRC Zeiss camera and Zen 2012 software. The images were analyzed using CellProfiler (version 2.1.1) and CellProfiler Analyst open-source software. Out of the 10,378 objects, 4970 (47,9%) were identified as TUNEL positive, and 5408 (52,1%) were identified as TUNEL negative. On average, the sensitivity and specificity values of the machine learning approach were 0.80 and 0.77, respectively. Image cytometry provides a quantitative analytical alternative to the more traditional qualitative methods more commonly used in studies.

Hepato- and nephroprotective effects of bradykinin potentiating factor from scorpion (Buthus occitanus) venom on mercuric chloride-treated rats.

Bioactive peptides such as bradykinin potentiating factor (BPF), have, anti-oxidative, anti-inflammatory, immunomodulatory and ameliorative effects in chronic diseases and play a potential role in cancer prevention. It is known that the liver and kidney accumulate inorganic mercury upon exposure, which often leads to mercury intoxication in these organs. In this study, we investigated the effect of bradykinin potentiating factor (BPF), a scorpion venom peptide, on mercuric chloride-induced hepatic and renal toxicity in rats. We used 20 adult male Albino rats divided into four equal groups: the first group was injected with saline (control); the second group was administered daily with mercuric chloride (HgCl2) for 2 weeks; the third group was administered with BPF twice weekly for 2 successive weeks, while the fourth group was exposed to BPF followed by HgCl2. We observed that HgCl2 treated rats had a significant increase in serum ALT, AST, ALP, creatinine and urea levels compared to control. Furthermore, HgCl2 treated rats showed a marked decrease in total proteins, albumin and uric acids compared to control. The previously studied parameters were not significantly changed in BPF pretreated rats compared to control. Moreover, a significant decrease in the activities of glutathione perioxidase (GSH), superoxide dismutase (SOD), and catalase (CAT), in addition to a significant increase in the level of malondialdehyde (MDA) were observed in hepatic and renal tissues of rats after HgCl2 treatment. In contrast, the HgCl2/BPF treated rats showed a significant elevation in the activity of GSH, SOD, and CAT accompanied with a significant regression in the level of MDA compared to the HgCl2 exposed rats. We conclude that treatment with BPF is a promising prophylactic approach for the management of mercuric chloride-induced hepato- and nephro-toxicities.

Influence of Sanao Tang on urine volume and expression of aquaporin 2 in rats with respiratory function impairment induced by passive smoking and lipopolysaccharide

OBJECTIVETo investigate the effect of Sanao Tang (SAT) on urine volume and the expression of aquaporin-2 (AQP2) in rats with lung dysfunction induced by passive smoking and lipopolysaccharide. METHODSTotally 45 healthy Specific pathogen Free Wistar Rats were randomized into 3 groups: normal control group, model group and SAT group. A rat model of respiratory dysfunction induced by exposure to cigarette smoking and lipopolysaccharide (LPS). Lavage of decoction of the Chinese medicine was performed for rats in the SAT group. Anires 2005 System was used to analyze the pulmonary function. Urine of rats was collected through metabolism cage method. Enzyme-linked immunosorbent assay (ELISA) was used to determine content of antidiuretic hormone (ADH), angiotensin II (Ang II), atrial natriuretic factor (ANP), endothelin 1 (ET-1), nitric oxide (NO) and prostaglandin E2 (PGE2) in serum, lung and kidney. The expression of AQP2 in rat renal tissue was determined with real-time fluorescence quantitative PCR (RT-PCR). RESULTS(a) In comparison with the normal control, It was found that enforced vital capacity (FVC), 1-second forced expiratory volume/forced vital capacity% (FEV1/FVC%), 24 h urine volume content of NO and PGE2 were decreased, while AQP2mRNA level and content of ADH, Agn II, ANP and ET-1 were increased in the model group with statistical significance (P < 0.05). (b) In comparison with the model group, It was found that FVC, FEV1, FEV1/FVC%, 24 h urine volume, content of PGE2 and NO decreased, while AQP2mRNA level, content of ANP, ADH and Ang II decreased in the SAT group with statistical significance (P < 0.05). CONCLUSIONSAT might effectively regulate the urine volume in the modeled rats; ADH, Ang II, ANP, ET-1, NO and PGE2 might play an important role in the regulation on urine volume by lungs. This might be the mechanisms underpinning the function of lung governing water passage in terms of the theory of Traditional Chinese Medicine.

Assessment of SGLT2 inhibitors on kidney morphology and albumin excretion in rats with experimental diabetes mellitus type 1

Background. Recently, as the therapy of diabetes mellitus, have been approved several members of a new class of drugs – SGLT2 inhibitors. SGLT2 inhibitors reduce glycemia and normalize renal perfusion. However renoprotective effect is still the subject of research.The aim of this study was to evaluate the effect of SGLT2 inhibitors on renal tissue of rats with experimental type 1 diabetes (DM).Materials and methods. This study was conducted on 22 white male rats at the age of 10 months with streptozotocin - induced diabetes. For the selection of the rats in the experiment were evaluated glycemia. The criteria for diabetes was blood glucose levels &gt; 7 mmol / L and / or a positive glucose tolerance test. Rats were divided into 3 groups: 1 group (healthy control) - a group of 10 animals without DM, 2 group - (diabetic control) 6 animals with DM receiving insulin NPH; 3 -6 animals with DM receiving insulin NPH and dapagliflozin 0.1 mg / kg for 4 weeks. At the end of the study, animals were kept in metabolic cages and 24 hour urine was collected for estimation of albuminuria. Next animals were removed from the experiment, and kidney tissue was sampled for morphological evaluation. Sections were stained with haematoxylin and eosin (H&amp;E), and PAS reaction stains for histopathological examination. Statistical significance of differences was assessed and linkages by standard methods of nonparametric statistics.Results. Sections of kidney from control group had classical structure of renal tissue. In contrast, histological sections from rats treated with insulin had showed cortical glomerulosclerosis, proliferation of mesangial cells and narrowed Bowman's spaces. In most of the proximal convoluted tubules was observed excessive hypertrophy, vacuolization and pyknotic nuclei. The kidneys of rats treated with dapagliflozin and insulin had a lower severity of degenerative processes compared with a group of rats treated with insulin. Many glomeruli had increased cellularity with normal Bowman’s spaces, while in the proximal convoluted tubules was observed weakly pronounced vacuolization and pyknotic nuclei and some less hypertrophy of tubular epithelium. Kidney sections of insulin-treated rats were showed signs of diffuse expansion of mesangial area with proliferation of mesangial cells and its PAS-positive matrix. While signs of mesangial expansion were absent in the dapagliflozin group. Analysis of the degree of glomerulosclerosis data was showed significant differences between the group of rats treated with dapagliflozin and insulin and the group of rats treated with insulin - 0.5 (0.4 - 0.6) and 1.1 (1.0 - 1.2), respectively ( p = 0.005). Furthermore, it revealed significant difference in percentage of mesangial area between group of rats treated with dapagliflozin and insulin and the group of rats treated with insulin - 28% (23 - 32) and 37% (33 - 41), respectively (p = 0.0082). Insulin-treated rats were showed significantly higher level of albuminuria compared with dapagliflozin-treated rats - 91.8 mg / 24h. (74.1 - 108.5) and 50.9 mg / 24 hr (41.3 - 60.2), respectively, (p = 0.012).Conclusions. Administration of dapagliflozin slows the progression of glomerulosclerosis and reduces the degree of its severity and the level of albuminuria, which may suggest a renoprotective properties.

Effect of penehyclidine hydrochloride pretreatment on Nrf2/HO-1 signaling pathway in renal tissues of rats with rhabdomyolysis-induced acute kidney injury

Objective To investigate the effect of penehyclidine hydrochloride (PHC) pretreatment on nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway in renal tissues of rats with rhabdomyolysis-induced acute kidney injury (AKI). Methods Thirty-six pathogen-free male Sprague-Dawley rats, weighing 200-220 g, were assigned into 3 groups (n=12 each) using a random number table: control group (group C), group AKI and PHC pretreatment group(group PHC). Rhabdomyolysis was induced by intramuscular injection of 50% glycerol 10 ml/kg in bilateral hindlimbs.PHC 0.2 mg/kg was injected intraperitoneally at 30 min before glycerol was injected intramuscularly in group PHC.At 1 and 6 h after glycerol injection, serum was collected for determination of blood urea nitrogen(BUN) and creatinine(Cr) concentrations, and bilateral kidneys were harvested for pathological examination and for determination of HO-1 activity and expression of Nrf2 mRNA and HO-1 mRNA (by quantitative real-time polymerase chain reaction), Nrf2 in nucleoprotein and total protein and HO-1 in total protein in renal tissues (by Western blot). The damage to the renal tubules was scored. Results Compared with group C, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly increased, the expression of Nrf2 in nucleoprotein and total protein and HO-1 in total protein was significantly up-regulated, and HO-1 activity was significantly increased in AKI and PHC groups, the expression of HO-1 mRNA was significantly up-regulated in group AKI, and the expression of Nrf2 mRNA and HO-1 mRNA was significantly up-regulated in group PHC (P<0.01 or 0.05). Compared with group AKI, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly decreased, the expression of Nrf2 in nucleoprotein and total protein and HO-1 in total protein was significantly up-regulated, and HO-1 activity was significantly increased in group PHC (P<0.01 or 0.05). Conclusion The mechanism by which PHC pretreatment attenuates rhabdomyolysis-induced AKI may be related to activation of Nrf2/HO-1 signaling pathway in renal tissues of rats. Key words: Penehyclidine hydrochloride; Rhabdomyolysis; Kidney; Nuclear factor erythroid 2-related factor 2; Heme oxygenase-1