Decellularization with triton X-100 provides a suitable model for human kidney bioengineering using human mesenchymal stem cells.

AIMS: Regeneration of discarded human kidneys has been considered as an ideal approach to overcome organ shortage for the end-stage renal diseases (ESRDs). The aim of this study was to develop an effective method for preparation of kidney scaffolds that retain the matrix structure required for proliferation and importantly, differentiation of human adipose-derived mesenchymal stem cells (hAd-MSCs) into renal cells. MAIN METHODS: We first compared two different methods using triton X-100 and sodium dodecyl sulfate (SDS) for human kidney decellularization; followed by characterization of the prepared human renal extracellular matrix (ECM) scaffolds. Then, hAd-MSCs were seeded on the scaffolds and cultured for up to 3 weeks. Next, viability, proliferation, and migration of seeded hAd-MSCs underwent histological and scanning electron microscopy (SEM) assessments. Moreover, differentiation of hAd-MSCs into kidney-specific cell types was examined using immunohistochemistry (IHC) staining and qRT-PCR. KEY FINDINGS: Our results indicated that triton X-100 was a more effective detergent for decellularization of human kidneys compared with SDS. Moreover, attachment and proliferation of hAd-MSCs within the recellularized human kidney scaffolds, were confirmed. Seeded cells expressed epithelial and endothelial differentiation markers, and qRT-PCR results indicated increased expression of platelet and endothelial cell adhesion molecule 1 (PECAM-1), paired box 2 (PAX2), and E-cadherine (E-CDH) as markers of differentiation into epithelial and endothelial cells. SIGNIFICANCE: These observations indicate the effectiveness of decellularization with triton X-100 to generate suitable human ECM renal scaffolds, which supported adhesion and proliferation of hAd-MSCs and could induce their differentiation towards a renal lineage.

Kidney dysfunction and oxidative stress in doxorubicin-induced nephrotic rat: Protective role of sesame oil.

Doxorubicin (DOX) is an antineoplastic agent which it's clinical use has been limited due to its major side effects including cardiotoxicity and nephrotic syndrome. Sesame oil (SO) is an important edible oil with many pharmacologic effects. The aim of the present study was to investigate the effect of SO against DOX-induced nephropathy in the rat. In this study, two doses of SO (3 and 6 mL/kg) were administrated orally for six consecutive weeks and DOX (mg/kg) was intravenously injected on the 4th day of the experiment. Blood and urine samples were collected on days 1, 14, 30, and 42 for subsequent measurement of biochemical parameters. The left kidneys were removed for subsequent assessment of total thiol content, malondialdehyde (MDA) concentration, and renal activities of catalase and superoxide dismutase enzymes. DOX caused significant proteinuria, hypoalbuminemia, and hyperlipidemia compared to control group. Significant decrease in antioxidant enzyme activities and total thiol contents and significant increase in MDA levels were also observed following DOX injection when compared to control group. Oral administration of SO significantly reversed DOX-induced proteinuria, hypoalbuminemia, and hyperlipidemia compared to DOX group. Furthermore, compared to DOX group, SO significantly increased total thiols content. MDA concentration significantly decreased following SO administration when compared to DOX group. The current study suggests that SO is able to improve kidney function as well as kidney tissue oxidative damage in DOX-induced nephrotic the rat.

Renoprotective Effect of Plantago major Against Proteinuria and Apoptosis Induced by Adriamycin in Rat.

OBJECTIVE: Adriamycin (ADR) is an important anti-cancer drug which can cause renal toxicity. Given the known anti-inflammatory and antioxidant effects of Plantago major (P. major), the aim of this study was to determine the effects of hydroalcoholic extract of P. major on ADR- induced nephropathy in rats. METHODS: Fifty male Wistar albino rats were randomly divided into 5 groups including: control, ADR (5 mg/kg), ADR + P. major (600 and 1200 mg/kg) and P. major (1200 mg/kg). The animals were treated with P. major extract for 5 consecutive weeks and ADR was intravenously injected on the 7th day of the study. Urine and serum samples were collected on days 0, 14, 21, 28, and 35 for the measurement of serum cholesterol and albumin levels and urine protein excretion rate. At the end of the study, the left kidneys were removed for apoptosis assessment. RESULTS: Administration of ADR significantly decreased serum albumin level and increased serum cholesterol and urine protein excretion rate as well as, apoptotic cell numbers compared to the control group (P < 0.001) while had no effect on glomerular filtration rate (P > 0.05). Treatment with P. major, in both 600 and 1200 mg/kg doses, increased serum albumin level and decreased serum cholesterol concentration, urine protein excretion rate and as well as the number of apoptotic cell compared to the ADR group (P < 0.001). CONCLUSION: Our results showed that the P. major extract effectively protects against ADR- induced nephropathy by reducing kidney apoptosis and improving renal functioning in rats.

Plantago major protects against cisplatin-induced renal dysfunction and tissue damage in rats.

The aim of the present study was to determine the effect of Plantago major (P. major) on cisplatin-induced kidney injury in the rat. Cisplatin was injected on the 6th day of the experiment. Animals were treated with P. major extract (300, 600, and 1200 mg/kg) and Vitamin E for five days before and two weeks after cisplatin administration. Cisplatin caused a significant decrease in glomerular filtration rate (GFR), urine osmolarity, and urinary excretion rate of potassium, but significant increase in the kidney index and histological damage compared with the control group. Administration of Vitamin E and P. major (300 and 600 mg/kg) significantly increased GFR compared to cisplatin group. Furthermore, urine osmolarity in Vitamin E and P. major (600 mg/kg) groups were significantly elevated compared to the cisplatin group. P. major (600 mg/kg) significantly increased the urinary excretion rate of potassium compared with cisplatin group. Furthermore, all doses of P. major and Vitamin E significantly attenuated the percentage of kidney tissue damage compared to the cisplatin group. However, only P. major (600 mg/kg) and Vitamin E treated rats showed a significant reduction in the kidney index. This study revealed that P. major extract in a dose-dependent manner provides protection against renal damage induced by cisplatin.

Renoprotective effect of Nigella sativa against cisplatin-induced nephrotoxicity and oxidative stress in rat.

Cisplatin is one of the important antineoplastic drugs. Its clinical use has been restricted due to severe kidney toxicity. Nigella sativa (N. sativa) is an herbaceous plant with many pharmacologic effects. In the present study, we evaluated the protective effects of aqueous-ethanolic extract of N. sativa and Vitamin E on cisplatin-induced nephrotoxicity in rats. Eighty male rats were divided into eight groups: control, cisplatin (6 mg/kg; ip), preventive Vitamin E (100 mg/kg), preventive N. sativa (100,200 mg/kg), preventive + treatment Vitamin E, and preventive + treatment N. sativa (100, 200 mg/kg). Duration of this study was 11 days and cisplatin was injected on the 6th day of the experiment. Tissue damage in all groups that received N. sativa extract and Vitamin E showed a significant improvement compared with the cisplatin group. In addition, serum and tissue total thiol content in preventive and preventive + treatment N. sativa groups showed significant increase compared with cisplatin group. There was no significant difference in serum malondialdehyde concentration of the control rats compared with the preventive and preventive + treatment N. sativa groups. N. sativa extract and viamin E improved the pathology and oxidative stress in the rat kidney. However, more studies are needed to determine the mechanism of action of N. sativa on cisplatin-induced kidney toxicity.

Effects of standardized Zataria multiflora extract and its major ingredient, Carvacrol, on Adriamycin-induced hepatotoxicity in rat.

BACKGROUND: Due to antioxidant effects of Zataria multiflora (ZM) and Carvacrol (CAR) in many cases and the prominent role of reactive oxygen species (ROS) in hepatotoxicity induced by Adriamycin (ADR), the aim of this study is to investigate the effects of ZM and CAR on ADR-induced hepatotoxicity. METHODS: Twenty four male Wistar rats were randomly divided into four groups including: 1)Control, 2)Adriamycin (ADR), 3,4) ZM + ADR and CAR + ADR that received ZM and CAR for 28 consecutive days. Blood samples were collected on the days 0, 14 and 28 to determine the alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. Also, the hepatic redox markers were evaluated. RESULTS: ADR significantly increased ALP, ALT and AST in comparison with the control (p < 0.05 - p < 0.001). In CAR + ADR group, the serum ALP, ALT and AST were significantly reduced compared to those of the ADR group (p < 0.01 to p < 0.001). Also, in ZM + ADR group, serum ALP and ALT compared to ADR was significantly reduced (p < 0.001). MDA level in the ADR group significantly increased compared to control (p < 0.01). The MDA level in ZM + ADR (p < 0.05) and CAR + ADR (p < 0.01) groups were significantly reduced compared to that of ADR. Thiol levels in ZM + ADR group significantly increased compared to the ADR group (p < 0.05). The activities of CAT in the ADR group was significantly reduced compared to control (p < 0.05) and increased in treatment groups in comparison with the ADR (p < 0.01). CONCLUSION: Long-term administration of ZM extract and CAR could reduce the oxidative damage in the rat liver induced by ADR through the strengthening of the antioxidant system.

The Preventive Effects of Standardized Extract of Zataria multiflora and Carvacrol on Acetaminophen-Induced Hepatotoxicity in Rat: - Zataria multiflora and Carvacrol and Hepatotoxicity.

OBJECTIVES: The hepatotoxicity induced by Acetaminophen (AAP) mostly mediated by effect on oxidative stress parameters. The Zataria multiflora (Z.M) is an herbal medicine with well-known antioxidant effect. The aim of this study is investigation of preventive effects of Z.M and Carvacrol (CAR) on AAP-induced hepatotoxicity in rats. METHODS: Rats were randomly divided into four groups including: 1) Control, 2) Acetaminophen (AAP), 3) and 4) CAR. The saline, Z.M (200 mg/kg) and CAR (20 mg/kg) were administrated orally for 6 days, after that AAP (600 mg/kg) was administrated in the 7th day. Blood sampling was performed on the first and last days. Also, the liver tissue was removed for evaluation of Malondyaldehide (MDA), Thiol content, Superoxide dismutase (SOD) and Catalase (CAT). Total Protein (tPro), Glutamic Oxaloacetic Transaminase (GOT), Glutamic Pyruvic Transaminase (GPT) and Alkaline Phosphatase (ALP) in liver tissue were evaluated. The changes (Δ) of enzymes activities were presented. RESULTS: The Δ GOT, Δ GPT and Δ ALP in CAR group significantly decreased compared to AAP group (P < 0.01 to P < 0.001) and Δ GPT in Z.M group was significantly reduced in comparison with AAP group (P < 0.05). Also, MDA, Thiol, SOD and CAT levels in treated groups were attenuated compared to AAP group (P < 0.05 to P < 0.001). CONCLUSION: Z.M and CAR have a powerful hepatoprotective effect. CAR is more effective than Z.M. Based on the results. Z.M and CAR could be potent supplementary agents against hepatotoxicity of AAP in patients.

Mixed hydroalcoholic extracts of Nigella sativa and Curcuma longa improves adriamycin-induced renal injury in rat.

Extracts of both Curcuma longa (CL) and Nigella sativa extract (NS) are reported to have protective effects on renal damage. In this study, we investigated the protective effect of a combination of NS and CL on Adriamycin (ADR)-induced renal damage. Forty eight rats were divided into six groups as: Control (CO), ADR, Vitamin C + ADR, CL + ADR, NS +ADR, and CL + NS + ADR. ADR was injected intravenously on the 7th day of the study. 24-hour urine and orbital blood samples were collected on day 0, 48 hr after ADR injection and at the end of weeks 2, 3, 4, and on the 35th day. Glomerular filtration rate (GFR) was calculated on each sample, and on the 35th day, renal index and histological changes were also evaluated. In the ADR-treated rats, significant renal pathological changes were demonstrated compared to CO group. The renal index and urine protein excretion significantly increased, and serum albumin and GFR in the ADR-treated rats were significantly decreased compared to CO group. In NS + ADR group, the serum albumin significantly decreased compared to ADR group. In CL + NS + ADR group, the urine protein excretion was lower than ADR group, and serum albumin concentration was significantly higher than ADR group. In addition, in CL + ADR and NS + ADR groups also, the urine protein was significantly lower compared to ADR group. This study shows that the mixed extracts of N. sativa and CL have positive synergistic effects on renal damage in nephropathy induced by ADR in rats.

Thymoquinone ameliorates renal damage in unilateral ureteral obstruction in rats.

BACKGROUND: It is well established that renin-angiotensin system (RAS) play a central role in pathophysiology of renal damage following unilateral ureteral obstruction (UUO). The present study investigated the effects of thymoquinone and RAS blockade on renal tissue damage and renal expression of angiotensin II after UUO in rats. METHODS: Forty male rats were divided to five groups: Sham-operated group, UUO group and rats with UUO treated with losartan, captopril and thymoquinone. The rats were evaluated two weeks after UUO by measuring renal oxidative stress, apoptosis and the expression of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein (MCP-1) and angiotensin II. RESULTS: Two weeks after UUO there was a significant increase in the number of renal apoptotic cells and renal malondialdehyde and TNF-α expression. In addition, renal total thiol content and the activity of antioxidant enzymes were significantly decreased in UUO group, compared with the sham group. Also, UUO was associated with upregulation in the expression of angiotensin II and MCP-1 in the obstructed kidney. Losartan, captopril and thymoquinone significantly improved oxidative damage, apoptosis and TNF-α expression and markedly decreased the upregulation of angiotensin II and MCP-1 compared with the UUO group. CONCLUSIONS: The current study suggests that thymoquinone is able to improve the UUO-induced renal tissue damage. These favorable actions of thymoquinone on UUO model in rat are comparable with the well-known RAS inhibitors captopril and losartan.

The role of local renin-angiotensin system on high glucose-induced cell toxicity, apoptosis and reactive oxygen species production in PC12 cells.

OBJECTIVES: Hyperglycemia, oxidative stress and apoptosis have key roles in pathogenesis of diabetic neuropathy. There are local renin-angiotensin systems (RASs) in different tissues such as neural tissue. Local RASs are involved in physiological and pathophysiological processes such as inflammation, proliferation and apoptosis. This study aimed to investigate the role of local renin-angiotensin system on high glucose-induced cell toxicity, apoptosis and reactive oxygen species (ROS) production in PC12 cells, as a cell model of diabetic neuropathy. MATERIALS AND METHODS: PC12 cells were exposed to a high glucose concentration (27 mg/ml), captopril (ACE inhibitor), telmisartan and losartan (AT1 antagonists), and also PD123319 (AT2 antagonist) were administered before and after induction of high glucose toxicity. Then cell viability was assessed by MTT assay and apoptotic cells and intracellular ROS production were detected by annexin V-propidium iodide and DCFDA, respectively, using flow cytometry. RESULTS: High glucose concentration decreased cell viability, and increased apoptotic cells. Intracellular ROS production was also increased. In PC12 cells pretreatment and treatment by the drugs showed a significant improvement in cell viability and reduced apoptosis in captopril, telmisartan and PD123319 but only captopril and telmisartan were able to reduce ROS production. Losrtan significantly lowered ROS but didn't show any improvements in cell viability and apoptotic cells. CONCLUSION: The results of the present study showed that RAS inhibitors reduced cell toxicity and apoptosis and ROS production was induced by high glucose. It may be suggested that local RAS has a role in high glucose toxicity.

Effect of Nigella sativa on ischemia-reperfusion induced rat kidney damage.

OBJECTIVES: There are a few previously reported studies about the effect of Nigella sativa oil on renal ischemia-reperfusion injury (IRI). The aim of the present study was to test the hypothesis whether pre- or post-treatment with N. sativa hydroalcoholic extract (NSE) would reduce tissue injury and oxidative damages in a clinically relevant rat model of renal IRI. MATERIALS AND METHODS: IRI was induced by clamping of bilateral renal arteries for 40 min fallowed by reperfusion for 180 min. NSE was prepared in a Soxhlet extractor and administrated with doses of 150 mg/kg or 300 mg/kg at 1 hr before ischemia induction (P-150 and 300) or at the beginning of reperfusion phase (T-150 and 300), via jugular catheter intravenously. The kidneys were then removed and subjected to biochemical analysis, comet assay or histopathological examination. RESULTS: The kidneys of untreated IRI rats had a higher histopathological score (P<0.001), while in P-150, as well as T-150 and T-300 groups tubular lesions significantly decreased (P<0.001). Pre- and post-treatment with NSE also resulted in a significant decrease in malondialdehyde (MDA) level (P<0.001) and DNA damage (P<0.001) that were increased by renal I/R injury. NSE treatment also significantly restore (P<0.01) the decrease in renal thiol content caused by IRI. CONCLUSION: The present study shows N. sativa extract has marked protective action against renal IRI, which may be partly due to its antioxidant effects.

Alcea rosea root extract as a preventive and curative agent in ethylene glycol-induced urolithiasis in rats.

INTRODUCTION: Alcea rosea L. is used in Asian folk medicine as a remedy for a wide range of ailments. The aim of the present study was to investigate the effect of hydroalcoholic extract of Alcea rosea roots on ethylene glycol-induced kidney calculi in rats. MATERIALS AND METHODS: Male Wistar rats were randomly divided into control, ethylene glycol (EG), curative and preventive groups. Control group received tap drinking water for 28 days. Ethylene glycol (EG), curative and preventive groups received 1% ethylene glycol for induction of calcium oxalate (CaOx) calculus formation; preventive and curative subjects also received the hydroalcoholic extract of Alcea rosea roots in drinking water at dose of 170 mg/kg, since day 0 or day 14, respectively. Urinary oxalate concentration was measured by spectrophotometer on days 0, 14 and 28. On day 28, the kidneys were removed and examined histopathologically under light microscopy for counting the calcium oxalate deposits in 50 microscopic fields. RESULTS: In both preventive and curative protocols, treatment of rats with hydroalcoholic extract of Alcea rosea roots significantly reduced the number of kidney calcium oxalate deposits compared to ethylene glycol group. Administration of Alcea rosea extract also reduced the elevated urinary oxalate due to ethylene glycol. CONCLUSION: Alcea rosea showed a beneficial effect in preventing and eliminating calcium oxalate deposition in the rat kidney. This effect is possibly due to diuretic and anti-inflammatory effects or presence of mucilaginous polysaccharides in the plant. It may also be related to lowering of urinary concentration of stone-forming constituents.

Evaluation of the effect of formic acid and sodium formate on hair reduction in rat.

Hirsutism is a common problem in dermatology that imposes high socioeconomical costs on medical care. Consequently, researchers are actively searching for cheaper and safer methods for therapeutic treatment. The objective of the present study is to evaluate formic oil, enriched from formic acid, for the removal of unwanted hair. In this study, 32 female rats (150-200 g) were randomly divided into four groups and maintained with normal water and food availability. A patch of skin was shaved on each rat for application of test solutions. The control group was treated with local once-daily applications of normal saline. The formic acid, acetic acid, and sodium formate groups were treated with once-daily applications of formic acid (pH 5.5), acetic acid (pH 5.5), or sodium formate, respectively. After 2 weeks, horizontally cut sample biopsies were removed, and the numbers of hair follicles were counted under high field microscopy by a specialist blinded to the treatments. Kolmogorov-Smirnov test results indicated a nonparametric distribution for the rat groups. ANOVA analysis indicated no statistically significant differences between groups (P < 0.05). There weren't any side effects or evidence for toxicity during the study period. However, hair follicle counts showed a descending order of control, acetic acid, formic acid, and sodium formate. Although the sodium formate group had the lowest hair follicle numbers, the difference was not statistically significant (P > 0.05). Formic acid was not effective in reducing hair follicle numbers in rats.

The preventive effect of N-butanol fraction of Nigella sativa on ethylene glycol-induced kidney calculi in rats.

BACKGROUND: The current study was carried out to determine whether the aqueous-ethanolic extract or the butanolic fraction of Nigella sativa (NS) seeds could prevent or reduce calculi aggregation in experimental calcium oxalate nephrolithiasis in Wistar rats. MATERIALS AND METHODS: Male Wistar rats were randomly divided into 5 groups: group A received tap drinking water for 28 days. Groups B, C, D and E received 1% ethylene glycol for induction of calcium oxalate (CaOx) calculus formation for 28 days. Rats in groups C, D and E also received aqueous-ethanolic extract of NS, N-butanol fraction and N-butanol phase remnant of NS, respectively, in drinking water at a dose of 250 mg/kg for 28 days. Urine concentration of oxalate, citrate, and calcium on days 0, 14, and 28, and also serum concentration of magnesium and calcium on days 0 and 28, were measured. On day 29, kidneys were removed for histopathologic study and examined for counting the calcium oxalate deposits in 10 microscopic fields. RESULT: Treatment of rats with N-butanol fraction and N-butanol phase remnant of NS significantly reduced the number and size of kidney calcium oxalate deposits compared with ethylene glycol group. Urinary concentration of oxalate in all experimental groups increased compared with control group on days 14 and 28, whereas the urine citrate concentration was lower in all experimental groups compared with control group on days 14 and 28. CONCLUSION: N-butanol fraction and N-butanol phase remnant of NS showed a beneficial effect on calcium oxalate deposition in the rat kidney. Therefore, the butanolic fraction of NS may be suggested for prevention of calcium oxalate calculi in humans.