The renoprotective effect of curcumin in cisplatin-induced nephrotoxicity

Abstract

The polyphenol curcumin has several pharmacological effects, including antioxidant, anti-inflammatory and anti-cancer features. In this study, we evaluated the effects of curcumin in cisplatin-induced nephrotoxicity in rats. Male Wistar rats were divided into four groups: (1) control; (2) cisplatin (7?mg/kg body weight, intraperitoneal as a single dose); (3) curcumin (100?mg/kg via gavage, for 10 days); and (4) cisplatin and curcumin. The cisplatin-treated rats exhibited kidney injury manifested by increased serum urea and creatinine (p?<?0.05). The kidney tissue from the cisplatin treated rats also exhibited a significant increase in the malondialdehyde (MDA) levels (p?<?0.05). The treatment with curcumin prevented a rise in the serum urea, creatinine and MDA levels when compared to the control group kidneys (p?<?0.05). The analysis the nicotinamide phosphoribosyltransferase (NAMPT) and sirtuin (SIRT) proteins (SIRT1, SIRT3 and SIRT4), which play important roles in the resistance to stress and the modulation of the threshold of cell death, showed similar trends (p?<?0.05). In the cisplatin-only treated rats, the induced renal injury decreased the levels of the NAMPT and SIRT proteins. Conversely, the curcumin increased the levels of the NAMPT and SIRT proteins in the cisplatin-treated rats (p?<?0.05). These data suggest that curcumin can potentially be used to reduce chemotherapy-induced nephrotoxicity, thereby enhancing the therapeutic window of cisplatin.     

Proanthocyanidin and fish oil potent activity against cisplatin-induced renal cell cycle arrest and apoptosis in rats

Abstract

Cisplatin is an effective chemotherapeutic agent that displays dose-limiting nephrotoxicity. In the present study, the efficacy of grape seed proanthocyanidin extract (GSPE: 100?mg/kg/day) and fish oil (FO: 5?mL/kg/day) against cisplatin-induced nephrotoxicity was evaluated in terms of DNA damage, histopathological changes and expression levels of molecular markers of apoptosis. The administration of cisplatin (CP) (7?mg/kg) results in an increasing percentage of S-phase, G2/M and apoptosis. Furthermore, CP induces apoptosis as indicated by an elevation of renal caspase-3 and reduction in the expression of BCL-2. In addition to occurred renal histopathological changes as manifested by tubular degeneration, degenerative glomerulus, necrotic tubular cells, and cell debris. On the other hand, the administration of GSPE or FO pre-cisplatin treatment can be ameliorated the current DNA cell cycle alterations by the restoration of expression of proteins related to apoptosis and reduced the undesirable renal histopathological changes. So, it can be concluded that the consumption of GSPE or FO might be useful for minimizing nephrotoxicity caused by cisplatin chemotherapy through their anti-apoptotic and antioxidant properties.     

卡维地洛改善顺铂致肾小管上皮细胞凋亡

目的 观察顺铂致急性肾衰竭(ARF)中肾小管上皮细胞凋亡情况及卡维地洛(carvedilol)对其影响,并探讨其作用机制。方法 雄性Wistar大鼠随机分为盐水对照组、顺铂组、卡维地洛对照组、卡维地洛治疗组。测定BUN、Scr、尿乙酰氨基葡糖苷酶(NAG)、肾组织丙二醛(MDA)与超氧化物歧化酶(SOD)的含量。苏木素-伊红染色(HE)观察肾脏病理改变。原位缺口末端标记法(TUNEL)与DNA电泳观察肾小管上皮细胞的凋亡。Western印迹检测caspase-3的蛋白表达。结果 顺铂组大鼠BUN、Scr、尿NAG升高;肾脏病理改变加重;大量的肾小管上皮细胞凋亡; MDA含量增加,SOD活性降低;caspase-3的蛋白表达增加。上述指标在卡维地洛治疗组均得到改善,BUN从(58.33±19.93)降至(28.74±19.62)mmol/L;Scr从(425.56±97.96)降至(253.90±134.87)μmol/L;NAG从(224.77±75.86)降至(137.52±26.38)U/L;MDA从(18.13±7.01)降至(9.74±1.68)nmol/mg蛋白;SOD从(30.05±12.20)升至(64.67±20.64)U/mg蛋白;caspase-3的蛋白表达从1.94±0.73降至1.25±0.52;细胞凋亡从(42.5±12.6)%降至(23.7±8.4)%;肾组织病理损害也明显改善。结论 肾小管上皮细胞凋亡是顺铂致ARF的重要原因之一。卡维地洛能减轻顺铂的肾毒性,降低肾小管上皮细胞凋亡,其机制可能与减少活性氧(ROS)的产生,部分抑制了caspase依赖的凋亡途径有关。     

Potential of morin and hesperidin in the prevention of cisplatin-induced nephrotoxicity

Oxidative stress is one of the important mechanisms of cisplatin-induced nephrotoxicity. Therefore, this study was designed to explore the potential protective effects of morin and/or hesperidin on oxidative stress in cisplatin-induced nephrotoxicity. This study was performed on 42 Wistar rats. Rats were divided into seven groups: control, morin, hesperidin, cisplatin, cisplatin?+?morin, cisplatin?+?hesperidin, and cisplatin?+?morin?+?hesperidin. Morin and/or hesperidin were given for 10 consecutive days by oral gavage and on the 4th day a single dose of cisplatin (7?mg/kg) was injected intraperitoneally. After administrations, on the 11th day of the experiment the animals were killed, and malondialdehyde (MDA), nitric oxide (NOx), glutathione (GSH) levels and myeloperoxidase (MPO), catalase (CAT), superoxide dismutase (SOD) activity were measured. Cisplatin-treated rats showed increased levels of MDA, and decreased levels of NOx also activity of CAT. Morin and/or hesperidin pretreatment prevent oxidative stress in kidney tissue, while they increase the NOx level, CAT activity, and decrease MPO activity. In conclusion, morin?+?hesperidin pretreatment may have a significant potential for protection of cisplatin-induced nephrotoxicity.     

Protective effect of pomegranate seed oil against cisplatin-induced nephrotoxicity in rat

Abstract

Purpose: Clinical use of cisplatin is limited by its nephrotoxicity. Cisplatin-induced nephrotoxicity is associated with an increase in oxidative stress, leading ultimately to kidney dysfunction. The aim of this study was to investigate the effect of pomegranate seed oil against nephrotoxicity induced by cisplatin in adult rats. Methods: Animals were divided into four groups. Group I received corn oil (1?mL/kg). Group II received cisplatin (8?mg/kg). Group III and IV received pomegranate seed oil (PSO) 0.4?mL/kg and 0.8?mL/kg one hour before cisplatin injection for 3 days, respectively. Blood samples were collected by cardiac puncture and used for measuring urea and creatinine concentration. Twenty-hour urine samples were collected to measure protein and glucose concentration. The right kidney fixed in formalin for histological examination and the left kidney was homogenized for measurement of malondialdehyde and total sulfhydryl groups. Results: A significant elevation of serum creatinine, urea, urinary glucose, protein concentrations, and non-significant decrease in total thiol content and increase in MDA level in kidney homogenates were observed in cisplatin-treated rats. Also cisplatin reduced animal’s body weight. Mild-to-moderate tubular cell necrosis, hyaline casts, and vascular congestion were observed in group II. PSO pre-treatment significantly decreased urinary protein, glucose, and serum creatinine concentration. PSO also caused a decrease in serum urea, renal MDA, and increase in thiol content, but the level of these parameters were not significant. Conclusion: The present results suggest that PSO is an effective agent for the prevention of cisplatin-induced renal dysfunction and oxidative damage in rat.     

Effect of free creatine therapy on cisplatin-induced renal damage

Cisplatin is one of the commonly used anticancer drugs and nephrotoxicity limits its use. The aim of this study is to investigate the possible protective effect of creatine supplementation on cisplatin-induced nephrotoxicity. Sixty male Sprague–Dawley rats were divided into three groups: Group I: Cisplatin (n?=?20) (7?mg/kg cisplatin intraperitoneal (i.p.) single dose), group II: Cisplatin?+?creatine monohydrate (n?=?20) (7?mg/kg cisplatin i.p. single dose and 300?mg/kg creatine p.o. daily for 30 days starting on first day of cisplatin injection), group III: Control group (n?=?20) (Serum physiologic, 2.5?mL/kg i.p.). Sacrifications were performed at first week and 30th day. Blood urea nitrogen (BUN) and serum creatinine levels, histopathological evaluation, mitochondrial deoxyribonucleic acid (mtDNA) common deletion rates, and body weights of rats were evaluated. A significant decrease in body weight, higher values of kidney function tests, histopathological scores, and mtDNA deletion ratios were observed in group I compared to control group at days 7 and 30 (p?p?=?0.931 and 0.084, respectively). Kidney function tests, histopathological scores, and mtDNA common deletion ratios were statistically better in group II than group I at 7th and 30th day (p?相似文献   

LIGHT／INF—γ基因配比转染HepG2细胞对其凋亡及Fas／FasL系统表达的影响

目的：观察脂质体介导的LIGHT和IFN—γ配比转染HepG2细胞后对其凋亡及Fas和FasL表达的影响。方法：将HepG2细胞分为LIGHT单独转染组、LIGHT和IFN—γ联合转染组和空白对照组，以脂质体为中介转染HepG2细胞；分别于转染后12、24和48h收集HepG2细胞，流式细胞术检测转染后HepG2细胞的凋亡率及Fas和FasL的表达。结果：LIGHT基因转染HepG2细胞能明显促进其凋亡，随时间延长凋亡率增加；Fas和FasL在HepG2细胞高表达，以Fas升高程度显著。结论：LIGHT和IFN—γ联合转染HepG2，其凋亡效果优于LIGHT单独转染，主要是通过上调Fas的表达来促进HepG2细胞凋亡。     

The efficacy of theophylline in preventing cisplatin-related nephrotoxicity in patients with cancer

Objective: Cisplatin is a potent antineoplastic agent used and its major limiting side effect is nephrotoxicity. The aims of the study are early detection of acute kidney injury (AKI) with biomarkers and investigation of the potential nephron-protective effects of theophylline. Methods: Glomerular filtration rates (GFR), neutrophil gelatinase-associated lipocalin (NGAL), cystatin C were measured at 5th day of treatment in all of the patients. In addition, these parameters were measured repeatedly after the administration of cisplatin, at 2nd hour, 5th and 20th days. Patients: Sixty patients who are planned to receive cisplatin for the first time were included in the study. Patients were divided into two groups as Group 1 (n?=?30) (standard treatment arm) and Group II (n?=?30) (theophylline arm). Results: In both groups after the administration of cisplatin, GFR showed a significant decrease within time (p?=?0.006). Urine NGAL levels were significantly high after 2?h of cisplatin administration (p?p?=?0.025). After 5 days of cisplatin administration, urine protein levels were significantly higher in both groups (p?Conclusion: Results showed that urine NGAL level is a superior biomarker compared to serum creatinine and serum cystatin C in the detection of early AKI. Theophylline was found not to bring a complete protection for the kidneys, but less nephrotoxicity was developed when compared to the group not receiving theophylline.     

姜黄素预处理对大鼠缺血再灌注肾小管上皮细胞凋亡的影响

目的：建立大鼠。肾脏缺血再灌注损伤（IRI）模型,观察姜黄素预处理对大鼠肾脏缺血再灌注肾小管上皮细胞凋亡的影响。方法：36只SD雄性大鼠随机分为3组,分别为假手术（Sham）组、肾脏缺血再灌注模型（IR）组、姜黄素预处理（CUR）组,每组12只。CUR组在缺血前2h给予姜黄素100mg／kg剂量溶于0．1％二甲基亚砜1ml中,注入腹腔。24小时后沿原切口进入,切除左。肾。肾组织用4％多聚甲醛固定24h,常规石蜡包埋切片。采用TUNEL法检测各组缺血肾小管上皮细胞凋亡。结果：与Sham组相比,IR组肾小管上皮细胞凋亡明显增加。与IR组比较,CUR组肾小管上皮细胞凋亡减少（P＜0．05）。结论：姜黄素预处理可减轻肾脏IRI的肾小管上皮细胞凋亡。     

Germ cell apoptosis induced by ureaplasma urealyticum infection

Aim: To study the effect of Ureaplasma urealyticum (UU) infection on germ cell apoptosis of male rats. Methods: Male rats were infected artificially with UU serotype 8 (T960). Morphological changes of germ cells in the seminiferous tubules and the lumen of the epididymides were observed under the light microscope. Fluorescence-conjugated polyclonal antibodies to Fas and Fas ligand (FasL) were used to localize Fas and FasL. TUNEL staining of germ cells and Sertoli cells was performed by the AKPase method. TUNEL-positive rate ( % positive cells) and TUNEL-positive area (area occupied by stained cells) were analysed by KS400 Image Analysis System. The DNA laddering analysis was performed by agarose gels electrophoresis. Results: In those rats infected with UU: (1) Exfoliated germ cells were dramatically increased. Many multinucleated giant cells were found in the seminiferous tubules and the lumen of the epididymides. (2) The number of TUNEL-positive cells and the TUNEL-positive area were significantly increased. (3) The expression of Fas and FasL in germ cells and Sertoli cells was up-regulated. (4) Discrete bands of fragmented DNA were found in the testicular cells. Conclusion: In male rats, germ cell apoptosis was increased in UU infection.     

安胎养血合剂对大鼠移植肾Fas/FasL基因表达及细胞凋亡的影响

目的：研究中药安胎养血合剂对移植肾肾小管上皮细胞凋亡及Fas/FasL基因表达的影响，探讨其免疫抑制作用的机制。方法：建立大鼠肾移植模型，随机分为3组，每组12对。分别为对照组：（Wistar→Wistar大鼠），给予生理盐水灌胃；生理盐水组：（SD→Wistar大鼠），给予生理盐水灌胃；安胎养血合剂组：（SD→Wistar大鼠），给予安胎养血合剂灌胃。于术后第5d处死动物取肾脏组织，通过TUNEL法检测凋亡指数和RT-PCR检测Fas/Fas-L的mRNA表达。结果：用TUNEL检测移植肾脏凋亡指数，生理盐水组凋亡细胞最多，对照组中凋亡细胞极少见，安胎养血合剂组和生理盐水组肾脏细胞凋亡指数比较差异有显著意义（P＜0.05）。生理盐水组肾小管上皮细胞Fas-L的mRNA表达显著增高，安胎养血合剂组Fas-L的mRNA表达显著减少，与生理盐水组之间比较有显著差异（P＜O.05）。结论：安胎养血合剂可以通过下调移植肾脏肾小管上皮细胞Fas-L的mRNA表达，来减少移植肾的细胞凋亡，起到免疫抑制剂的作用。     

姜黄素对人肾癌ACHN细胞放疗增敏作用的实验研究

目的研究姜黄素对人肾癌ACHN细胞放射增敏作用并探讨其作用机制。方法不同浓度姜黄素作用于ACHN人肾癌细胞24h后,MTT法检测姜黄素药物毒性;克隆形成实验观察其对放射敏感性的影响;流式细胞术检测姜黄素诱导细胞的凋亡率、细胞周期分布。结果姜黄素对人肾癌ACHN细胞有明显的抑制作用,可引起细胞的凋亡,且存在剂量和时间依赖;较低浓度的姜黄素（5μmol和10μmol）即可降低放射后ACHN细胞的克隆形成率,其放射增敏比分别为1．61及2．36;姜黄素及姜黄素联合辐射组的细胞周期阻滞在辐射敏感时相G2／M期;结论低剂量的姜黄素对人肾癌ACHN细胞有放射增敏作用,其机制可能与其引起细胞的凋亡增加,细胞周期阻滞有关。     

Diabetes and renal tubular cell apoptosis

Apoptosis contributes to the development of diabetic nephropathy, but the mechanism by which high glucose induces apoptosis is not fully understood. Apoptosis of tubular epithelial cells is a major feature of diabetic kidney disease, and hyperglycemia triggers the generation of free radicals and oxidant stress in tubular cells. Hyperglycemia and high glucose in vitro also lead to apoptosis, a form of programmed cell death. High glucose similar to those seen with hyperglycemia in people with diabetes mellitus, lead to accelerated apoptosis, a form of programmed cell death characterized by cell shrinkage, chromatin condensation and DNA fragmentation, in variety of cell types, including renal proximal tubular epithelial cells.     

姜黄素抑制肾癌ACHN细胞增殖及促凋亡的实验研究

目的观察姜黄素对人肾癌ACHN细胞株增殖及细胞凋亡的影响,探讨姜黄素诱导ACHN细胞株凋亡的作用机制。方法不同浓度姜黄素作用人肾癌ACHN细胞24 h后,应用MTT比色法检测姜黄素对人肾癌ACHN细胞的增殖抑制率;流式细胞术检测姜黄素诱导细胞的凋亡率;RT-PCR检测姜黄素对ACHN细胞Bcl-2、Bax、NF-κBP65 mRNA表达的影响;Western blot方法检测其对细胞Bcl-2、Bax、NF-κBP65I、κB蛋白表达的影响。结果姜黄素对人肾癌ACHN细胞有明显的抑制作用,可引起细胞凋亡,并且存在剂量和时间依赖;不同浓度姜黄素作用细胞24 h后,Bcl-2、NF-κBP65 mRNA水平下降,Bax mRNA水平升高(P0.05),Bcl-2、NF-κBP65蛋白表达量下降,BaxI、κB蛋白表达量升高(P0.05)。结论姜黄素通过上调IκB,下调NF-κB活性,调控凋亡基因Bcl-2/Bax的表达,抑制人肾癌ACHN细胞的增殖,诱导人肾癌ACHN细胞的凋亡。     

Melatonin decreases apoptosis and expression of apoptosis-associated proteins in acute puromycin aminonucleoside nephrosis.

BACKGROUND: The anti-apoptotic properties of melatonin have been demonstrated previously in several in vivo and in vitro studies. Previous reports have shown increased apoptosis during puromycin aminonucleoside nephrosis (PAN). The aim of this study was to determine if melatonin (MEL) can prevent apoptosis and modify oxidative stress, an apoptosis inducer, in this experimental model. METHODS: Rats were injected intraperitoneally with puromycin aminonucleoside. In addition, by the intragastric route they received 1 mg/kg/day of MEL or vehicle 3 days before puromycin injection and throughout the experiment. Animals were sacrificed at weeks 1 and 2 of nephrosis and frozen renal sections were studied for apoptosis by TUNEL, for apoptosis-associated proteins by monoclonal and polyclonal antibodies, and for superoxide anion (O(2)(-)) by a histochemical method. Nitric oxide (NO), malondialdehyde (MDA) and reduced glutathione (GSH), and the activities of superoxide dismutase (SOD) and catalase were measured in homogenized kidney tissue by appropriate biochemical and enzymatic methods. RESULTS: Increases in apoptosis, p53, Fas and Fas-ligand were observed in nephrotic animals. MEL treatment decreased apoptosis at weeks 1 and 2 in the glomerular, interstitial and tubular compartments. This was accompanied by decreased expression of p53 (glomerulus, week 1; tubules, weeks 1 and 2), Fas (glomerulus and interstitium, week 2; tubules, weeks 1 and 2) and Fas-ligand (interstitum and tubules, week 2). Increased expression of Bcl-2-positive cells was observed at week 2 in all renal compartments in MEL-treated animals. High levels of O(2)(-) and NO generation and lipid peroxidation (MDA) were found in nephrotic animals. SOD and GSH remained unchanged, and only decreased catalase activity (week 1) was observed in PAN animals. Tendencies toward decreased values of O(2)(-) and MDA content along with recovery of catalase activity (week 1) were observed in MEL-treated nephrotic animals, but were insignificant in magnitude. MEL, however, did significantly downregulate pro-apoptotic genes and upregulated anti-apoptotic genes. CONCLUSIONS: The data demonstrate that, in PAN, melatonin has anti-apoptotic effects, which might in part be independent of the modulation of the oxidative status.     

单侧输尿管梗阻对对侧肾小管上皮细胞凋亡影响的研究

目的探讨单侧输尿管梗阻对对侧肾小管上皮细胞凋亡的影响。方法新西兰大白兔48只,随机建立右侧输尿管完全梗阻4周和8周组;部分梗阻4周和8周组;假手术4周和8周组动物模型,每组8只。分别于梗阻4周、8周时取对侧肾脏,称重后利用TUNEL法检测对侧肾小管上皮细胞凋亡,并行HE染色。结果梗阻4周、8周均引起对侧肾实质代偿性增生,与梗阻程度及时间正相关（P〈0.05）。梗阻4周组HE染色见少数肾小管扩张,腔内存在坏死脱落细胞,以完全梗阻明显,但8周较4周相应各组无明显变化。TUNEL法检测完全梗阻、部分梗阻及假手术4周组对侧近曲小管上皮细胞凋亡指数分别为（6.02±1.21）%、（3.87±1.70）%、（2.32±1.39）%;远曲小管上皮细胞凋亡指数分别为（12.08±2.33）%、（7.22±3.35）%、（3.38±2.34）%;8周组对侧近曲小管上皮细胞凋亡指数分别为（7.00±2.19）%、（4.47±2.11）%、（2.29±0.85）%;远曲小管上皮细胞凋亡指数分别为（13.02±3.16）%、（8.11±2.99）%、（3.63±1.96）%。相应各组组间比较均有统计学意义（P〈0.05）,但8周与4周相应各组比较均无统计学意义（P〉0.05）。结论单侧输尿管梗阻早期可引起对侧肾小管上皮细胞损害,但随梗阻时间的延长并未进一步加重。     

The effect of alpha-melanocyte-stimulating hormone on renal tubular cell apoptosis and tubulointerstitial fibrosis in cyclosporine A nephrotoxicity

BACKGROUND: The pathogenesis of cyclosporine A (CsA)-induced nephrotoxicity has been known to be secondary to hemodynamic changes, but increasing evidence indicates that CsA has a direct toxicity to renal tubular cells, leading to their apoptosis and tubulointerstitial fibrosis. This study evaluated the mechanism for CsA-induced tubular cell apoptosis, tubulointerstitial fibrosis and its associated proteins, and the therapeutic effects of alpha-melanocyte-stimulating hormone (MSH) on them. METHODS: Male Sprague-Dawley rats fed with a low-sodium diet were divided into three treatment groups: group A (vehicle-injected group), group B (CsA 15 mg/kg-injected group), and group C(CsA+alpha-MSH-injected group). After 42 days, creatinine clearance; blood CsA level; apoptosis; inflammation and tubulointerstitial fibrosis in renal tissue; and the expression of Bax, Bcl2, Fas, FasL, and transforming growth factor (TGF)-beta protein were determined. RESULTS: CsA-induced tubular cell apoptosis; cellular infiltration; and increase of Fas, Bax, TGF-beta protein expression with significant tubulointerstitial fibrosis, and reduced Bcl2 protein expression. alpha-MSH treatment prevented the Bax and TGF-beta protein increase and induced Bcl2 protein increase, together with reduction of apoptosis, inflammation, and tubulointerstitial fibrosis. CONCLUSIONS: These findings suggest that chronic CsA nephrotoxicity is related to Bax and Bcl2-related apoptosis pathways, and that alpha-MSH can attenuate the CsA-induced tubulointerstitial fibrosis as well as tubular cell apoptosis.     

血清抗精子抗体阳性对青春期雄性大鼠睾丸组织及睾丸生殖细胞中Fas/Fas-L凋亡途径的影响

目的:通过人工免疫雄性大鼠获得抗精子抗体(AsAb)介导的血清AsAb阳性的免疫性不育大鼠动物模型,观察AsAb对青春期大鼠睾丸组织及睾丸生殖细胞中Fas/Fas-L凋亡途径的影响。方法:5周龄(青春期)雄性Wistar大鼠30只,其中10只处死,取精子制备精子悬液免疫大鼠,余20只动物随机分为实验组(10只)和对照组(10只),4周后摘取睾丸。光镜观察睾丸组织改变,免疫组化法检测Fas、Fas-L和Caspase-3蛋白的表达。结果:实验组睾丸组织切片呈凋亡样改变,实验组Fas、Fas-L及Caspase-3蛋白的OD值(176.97±4.58,187.52±7.76,157.65±7.38)较对照组(161.87±5.37,150.27±8.65,120.37±6.76)显著增高(P<0.01)。结论:同种精子免疫大鼠,可成功制作AsAb介导的免疫性不育模型;AsAb影响雄性大鼠的生育力,其机制可能与Fas/Fas-L凋亡途径中Fas、Fas-L和Caspase-3蛋白的表达升高有关。