右归丸对激素抵抗型肾病综合征大鼠肾脏组织中JNK/p38信号通路的影响

Effect of Yougui Pill on JNK/p38 Signaling Pathway in kidney Tissue of Rats with Hormone Resistance Nephrotic Syndrome

目的探讨右归丸治疗激素抵抗型肾病综合征的疗效及可能的机制研究。方法60只Wisar大鼠随机分为正常组(Control)、模型组(一次性尾静脉注射阿霉素制备)、泼尼松治疗组(6.3 mg·kg^-1)、泼尼松联合右归丸低(TL,6.3/2800 mg·kg^-1)、中(TM,6.3/5600 mg·kg^-1)、高(TH,6.3/11600 mg·kg^-1)剂量治疗组,每组10只大鼠,连续给药6周。给药2周、4周和6周测定大鼠体重、24 h尿蛋白含量;末次给药结束后,检测血脂、肾功能有关指标;采用HE、Masson、PAS染色观察大鼠肾组织病理形态学变化情况;Tunel染色观察大鼠肾组织细胞凋亡情况;免疫印迹法检测肾组织中纤维化蛋白、JNK/p38信号通路及凋亡有关蛋白表达情况。结果Model组2只死亡,TL、Prednisone组各死亡1只大鼠。给药2周Model组体重降低;给药4周、6周,与Model组、Prednisone组相比,激素右归丸联合治疗各组体重升高(P<0.05)。与Control组相比,Model组24 h尿蛋白量升高,TP、ALB水平降低,TC、TG、Scr、BUN、LDH水平升高,肾小球硬化评分、肾小管间质损伤评分、胶原纤维比例升高,肾组织细胞凋亡率升高,JNK、p38蛋白磷酸化水平升高,TGF-β1、CTGF、Caspase-3、Bax蛋白表达升高,Bcl-2蛋白表达降低(P<0.05)。与Model、Prednisone组相比,TL组、TM组、TH组24 h尿蛋白量降低,TP、ALB水平升高,TC、TG、Scr、BUN、LDH水平降低,肾小球硬化评分、肾小管间质损伤评分、胶原纤维比例降低,肾组织细胞凋亡率降低,JNK、p38蛋白磷酸化水平降低,TGF-β1、CTGF、Caspase-3、Bax蛋白表达降低,Bcl-2蛋白表达升高(P<0.05)。结论右归丸可缓解SRNS大鼠肾组织损伤,保护肾功能,这可能与右归丸抑制JNK/p38信号通路,降低肾组织细胞凋亡有关。

Objective To investigate the efficacy and possible mechanism of Yougui Pill in the treatment of hormoneresistant nephrotic syndrome.Methods 60 Wisar rats were randomly divided into control group,model group(preparation of adriamycin by one-off tail vein injection),prednisone treatment group(6.3 mg·kg^-1),prednisone combined with Yougui Pill low(TL,6.3/2.8 mg·kg^-1),medium(TM,6.3/5.6 mg·kg^-1),high(TH,6.3/11.6 mg·kg^-1)dose treatment groups,with 10 rats in each group.The rats were administered continuously for 6 weeks.The body weight and24-hour urinary protein content of rats were measured at 2,4 and 6 weeks after administration,after the last administration,blood lipid and renal function were measured,HE,Masson and PAS staining were used to observe the pathological changes of rat kidney,Tunel staining was used to observe the apoptosis of rat kidney cells,and the expressions of JNK/p38 signaling pathway and apoptosis-related proteins in kidney tissues were detected by Western blot.Results Two rats died in Model group and one rat died in TL and Prednisone groups respectively.The weight of model group decreased after 2 weeks of administration,after 4 and 6 weeks of administration,compared with Model group and Prednisone group,the weight of hormone and Yougui Pill combined treatment groups increased(P<0.05).Compared with the control group,the 24-hour urinary protein increased,the levels of TP and ALB decreased,the levels of TC,TG,Scr and BUN increased,the glomerulosclerosis score,tubulointerstitial injury score,collagen fibers ratio increased,the apoptotic rate of renal cells increased,the phosphorylation levels of JNK and p38 protein increased,the expressions of TGF-β1,CTGF,caspase-3 proteins increased in the model group,and the expression of Bcl-2 protein decreased(P<0.05).Compared with model group and prednisone group,in TL group,TM group and th group,24-hour urine protein decreased,TP and ALB levels increased,TC,TG,SCR,bun and LDH levels decreased,glomerulosclerosis score,renal tubulointerstitial injury score,collagen fiber ratio decreased,apoptosis rate of renal tissue decreased,JNK and p38 protein phosphorylation level decreased,TGF-β1,CTGF,caspase-3 and Bax protein expression decreased,bcl-2 protein expression decreased(P<0.05).Conclusions Yougui Pill can alleviate renal tissue damage and protect renal function in SRNS rats,which may be related to Yougui Pill inhibiting JNK/p38 signaling pathway and reducing renal cell apoptosis.