基于线粒体自噬途径探讨黄芪甲苷对阿霉素肾病大鼠的肾保护作用  

Renal Protective Effect of Astragaloside IV on Adriamycin-Induced Nephropathy in Rats Based on Mitochondrial Autophagy Pathway

作　　者：梁国强[1] 倪道磊[1] 周岚[1] 任燕[1] 宋永亮[1] 金伟民 蒋春波[1] LIANG Guoqiang;NI Daolei;ZHOU Lan;REN Yan;SONG Yongliang;JIN Weimin;JIANG Chunbo(Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,Jiangsu,China)

机构地区：[1]南京中医药大学附属苏州市中医医院,江苏苏州215009

出　　处：《中华中医药学刊》2022年第9期35-39,I0018,I0019,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074368);江苏省高层次卫生人才“六个一工程”拔尖人才科研项目(LGY2020046);苏州市第五批姑苏卫生拔尖人才项目(GSWS2019063);苏州市第九批姑苏卫生重点人才项目(GSWS2022107)。

摘　　要：目的从线粒体自噬角度探讨黄芪甲苷对阿霉素肾病大鼠的肾保护作用,进而阐释中药黄芪“扶补托毒”的护肾机理。方法SD大鼠尾静脉两次注射阿霉素(Adriamycin,ADR)建立ADR肾病大鼠模型。以随机数字法将模型大鼠分为模型组、西药组(洛丁新,0.9 mg·kg^(-1)·d^(-1)),黄芪甲苷低(20 mg·kg^(-1)·d^(-1))、中(40 mg·kg^(-1)·d^(-1))、高(80 mg·kg^(-1)·d^(-1))剂量组,每组8只。另选取8只同龄大鼠为正常组。各组大鼠灌胃给予相对应药物6周,每日1次,正常组与模型组予等容积生理盐水。给药干预后,生化法检测大鼠24 h定量尿蛋白及血清白蛋白(ALB)、丙氨酸氨基转移酶(ALT)和血清肌酐(Scr)、尿素氮(BUN)水平。苏木精-伊红染色法(HE)观察大鼠肾脏病理变化以及采用透射电镜观察肾组织线粒体结构、足细胞形态、数目的变化等。酶联免疫吸附剂法(ELISA)测定试剂盒检测肾组织白细胞介素-1β(IL-1β),活性氧(ROS)水平以及三磷酸腺苷(ATP)含量和线粒体膜电位(冰冻切片)采用化学荧光法检测。Real-time PCR法检测各组肾组织线粒体DNA(mitochondrial DNA,mt DNA)及核基因18srRNA的含量。Western blot法检测肾脏组织中线粒体自噬蛋LC3、Beclin-1和NLRP3、Caspase-1的蛋白表达。结果与正常组比较,模型组定量24 h尿蛋白,血清ALT、Scr和BUN水平,肾组织IL-1β水平以及LC3、Beclin-1和NLRP3、Caspase-1蛋白表达均显著性升高(P<0.05);血清ALB水平,肾组织ATP水平、mt DNA/18srRNA、膜电位显著降低(P<0.05);且病理结构检测结果显示其肾实质存在损伤病理变化,以及肾组织线粒体发生部分崩解和足细胞足突发生部分融合现象等。与模型组比较,西药组和黄芪甲苷中、高剂量组均能不同程度的改善相关指标,且黄芪甲苷高剂量减轻肾实质病理改变和线粒体微结构损伤最为明显。结论黄芪甲苷能够降低阿霉素肾病大鼠的蛋白尿,改善肾组织线粒体等病�Objective To explore the renal protective effect of astragaloside IV on adriamycin(ADR)-induced nephropathy in rats from the perspective of mitochondrial autophagy, and then to explain the renal protective mechanism of “supporting and tonifying for expelling toxin” of Huangqi(Astragali Radix).Methods ADR was injected twice into the tail vein of SD rats to establish the rat model of ADR-induced nephropathy.The model rats were randomly divided into model group, western medicine group(lotensin, 0.9 mg·kg^(-1)·d(-1)) and astragaloside IV low(20 mg·kg^(-1)·d(-1)),medium(40 mg·kg^(-1)·d(-1)) and high(80 mg·kg^(-1)·d(-1)) dose groups, with 8 rats in each group.Another 8 rats of the same age were selected as the normal group.Rats in each group were given the corresponding drugs by gavage once a day for 6 weeks, and the normal group and model group were given the equal volume normal saline.After administration, the levels of 24-hour quantitative urinary protein, serum albumin(ALB),alanine aminotransferase(ALT),serum creatinine(Scr) and urea nitrogen(BUN) were detected by automatic biochemical analyzer.The pathological changes of rat kidney were observed by hematoxylin eosin staining(HE),and the changes of mitochondrial structure, podocyte morphology and number were observed by transmission electron microscope.ELISA kit was used to detect interleukin-1β(IL-1β) in renal tissue.The levels of reactive oxygen species(ROS),adenosine triphosphate(ATP) and mitochondrial membrane potential(frozen section) were detected by chemical fluorescence method.The contents of mt DNA gene and nuclear gene 18 srRNA in renal tissues were detected by PCR.The protein expressions of mitochondrial autophagy LC3,beclin-1,NLRP3 and caspase-1 were detected by Western blot.Results Compared with those of the normal group, 24-hour urinary protein, serum levels of ALT,Scr and BUN and renal IL-1β were quantified in the model group and the expressions of LC3,beclin-1,NOD-like receptor protein 3(NLRP3) and caspase-1 were increased significant

关 键 词：黄芪甲苷 阿霉素肾病 自噬 线粒体 

分 类 号：R285.5[医药卫生—中药学]