柴胡皂苷A对糖尿病大鼠肾结构和功能的保护作用及机制研究  

Protective Effects of Saikosaponin A on Renal Structure and Function in Rats with Diabetes Mellitus and Its Mechanism

作　　者：张晓利[1] 胡威利 吴银[1] 孙昆[2] 周一龙[2] 路芳[1] ZHANG Xiao-Li;HU Wei-Li;WU Yin;SUN Kun;ZHOU Yi-Long;LU Fang(Dept.of Nephropathy and Rheumatism,the Third Affiliated Hospital of Xinxiang Medical College,Xinxiang 453000 Henan,China;Dept.of Blood Purification,the Third Affiliated Hospital of Xinxiang Medical College,Xinxiang 453000 Henan,China)

机构地区：[1]新乡医学院第三附属医院肾病风湿科,河南新乡453000 [2]新乡医学院第三附属医院血液净化室,河南新乡453000

出　　处：《广州中医药大学学报》2020年第7期1347-1353,共7页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：河南省科技计划项目(编号:162102310103)。

摘　　要：【目的】观察柴胡皂苷A对糖尿病大鼠肾结构和功能的保护作用及机制研究。【方法】分别建立糖尿病大鼠模型和高糖诱导的肾小管上皮细胞NRK-52E模型,应用柴胡皂苷A进行干预。检测糖尿病模型大鼠24 h尿蛋白(24hUP)、血清肌酐(SCr)和尿素氮(BUN),计算肾脏肥大指数;苏木素—伊红(HE)染色法观察大鼠肾脏病理损伤情况;酶联免疫吸附分析(ELISA)检测肾脏组织、NRK-52E细胞中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)含量;逆转录—聚合酶链反应(RT-PCR)、蛋白免疫印迹(Western Blot)法分别检测大鼠肾脏组织、NRK-52E细胞中异柠檬酸脱氢酶2(IDH2)、锰超氧化物歧化酶(MnSOD)和沉默信息调节因子3(SIRT3)mRNA和蛋白的表达水平。【结果】与正常组比较,模型组大鼠24hUP、SCr、BUN水平和肾脏肥大指数均显著增加(P<0.01),可见明显的肾组织损伤;与模型组比较,柴胡皂苷A中、高剂量组大鼠24hUP、SCr、BUN水平和肾脏肥大指数均显著降低(P<0.05或P<0.01),受损的肾组织得到明显改善。与正常组比较,模型组大鼠肾脏组织、NRK-52E细胞中SOD、GSH-Px活性及IDH2、MnSOD、SIRT3 mRNA和蛋白表达水平显著下降,MDA含量显著升高(P<0.01);与模型组比较,柴胡皂苷A中、高剂量组大鼠肾脏组织、NRK-52E细胞中SOD、GSH-Px活性及IDH2、MnSOD、SIRT3 mRNA和蛋白表达水平升高,MDA含量下降(P<0.05或P<0.01)。【结论】柴胡皂苷A可有效改善糖尿病大鼠肾结构和功能损伤,其机制可能与增强高糖状态下肾小管上皮细胞的抗氧化能力、减轻氧化应激反应有关。Objective To observe the protective effects of saikosaponin A on renal structure and function in rats with diabetes mellitus and its mechanism.Methods The diabetes mellitus rats model and high-glucose-induced renal tubular epithelial cell(NRK-52E)model were established,and were intervened by saikosaponin A.The 24-hour urine protein(24hUP),serum creatinine(SCr)and blood urea nitrogen(BUN)were measured,and renal mass index was calculated.The pathological features of rat renal tissue were observed by hematoxylin-eosin(HE)staining method.The activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),and content of malondialdehyde(MDA)in rat renal tissue and NRK-52E cells were detected by enzyme-linked immunosorbent assay(ELISA),and the mRNA and protein levels of isocitrate dehychogenase 2(IDH2),manganese-containing superoxide dismutase(MnSOD),and silent information regulator 3(SIRT3)were detected by reverse transcription polymerase chain reaction(RT-PCR)and Western blotting method,respectively.Results Compared with the normal group,the levels of 24hUP,SCr and BUN,as well as renal mass index were increased(all P<0.01)in the model group,and obvious renal tissue injury was seen.Compared with the model group,the levels of 24hUP,SCr and BUN,as well as renal mass index were decreased(P < 0.05 or P < 0.01)in the middle-,high-dosesaikosaponin A group, and obvious renal tissue injury was improved. Compared with the normal group, theactivity of SOD and GSH-Px,and the mRNA and protein expression levels of IDH2,MnSOD and SIRT3 in ratrenal tissue and NRK-52E cells were decreased, and MDA content was increased in the model group(all P <0.01). Compared with the model group,the activity of SOD and GSH-Px,and the mRNA and protein expressionlevels of IDH2,MnSOD and SIRT3 in rat renal tissue and NRK-52E cells were increased,and MDA content wasdecreased in the middle-,high-dose saikosaponin A group(P < 0.05 or P < 0.01). Conclusion Saikosaponin Ahas obvious effects on improving injury of renal structure and function in rats

关 键 词：柴胡皂苷A 糖尿病 氧化应激 大鼠 细胞模型 

分 类 号：R285.5[医药卫生—中药学]