摘要
【目的】探讨丹参多酚酸盐对小鼠糖尿病肾病的干预作用及机制。【方法】将60只C57BL/6小鼠随机分为正常组、模型组、贝那普利组、丹参多酚酸盐组,每组15只。除正常组,其他组别采用腹腔注射链脲佐菌素(STZ)的方法复制糖尿病肾病小鼠模型。造模成功后随机分为模型组、贝那普利组、丹参多酚酸盐组,丹参多酚酸盐组、贝那普利组分别灌胃相应的药物,正常组和模型组灌胃等体积蒸馏水,共4周。给药结束后,检测24 h尿蛋白定量(UTP),血清空腹血糖(FBG)、尿素氮(BUN)、血清肌酐(SCr),酶联免疫吸附分析(ELISA)检测小鼠肾组织中超氧化歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)水平,分别采用过碘酸六胺银(PASM)染色和Masson染色法观察肾脏病理学变化,采用蛋白免疫印迹(Western Blot)法分别检测小鼠肾脏组织中p-Smad2/3、Smad2/3、Smad7蛋白表达水平,逆转录-聚合酶链反应(RT-PCR)法检测小鼠肾脏组织中Smad7、Smad2/3、TGF-β1 m RNA表达水平。【结果】与正常组比较,模型组FBG、UTP均显著增加(P<0.05),肾脏组织可见明显病理改变,肾脏组织中的MDA含量显著升高,SOD、GSH-Px活性下降,p-Smad2/3/Smad2/3比值,Smad2/3、TGF-β1 m RNA表达水平明显升高,Smad7蛋白与m RNA表达水平明显下降(均P<0.05);与模型组比较,丹参多酚酸盐组、贝那普利组UTP降低(P<0.05),受损的肾组织得到改善,肾组织中MDA含量下降,SOD、GSH-Px活性升高,p-Smad2/3/Smad2/3比值,Smad2/3、TGF-β1 m RNA表达水平显著降低,Smad7蛋白与m RNA表达水平升高(均P<0.05)。【结论】丹参多酚酸盐可明显改善糖尿病肾病小鼠肾功能,其机制可能与抑制氧化应激反应和调控TGF-β1/Smad信号通路,从而减轻肾纤维化有关。
Objective To explore the intervention effects of salvianolate on diabetic nephropathy in mice and its mechanism.Methods Sixty C57BL/6 mice were randomly divided into normal group,model group,benazepri group,salvianolate group,15 mice in each group.Apart from the normal group,the mice in the other groups were induced into model of diabetic nephropathy by intraperitoneal injection of streptozotocin(STZ).After successful modeling,benazepril group and salvianolate group were given intragastric administration of corresponding drug,respectively,and the normal group and model group were given intragastric administration of the same volume of distilled water.The treatment lasted for 4 weeks.After medication,24-hour urine protein quantification(UTP),fasting blood glucose(FBG),blood urea nitrogen(BUN),serum creatinine(SCr)were detected,the levels of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)in mouse renal tissue were detected by enzyme-linked immunosorbent assay(ELISA),the pathological changes of the mouse renal tissue were observed by periodic acid methena-mine silver(PASM)and Masson staining methods,the protein expression levels of p-Smad2/3,Smad2/3,Smad7 in mouse renal tissue were detected by Western blotting assay,and the m RNA levels of Smad7,Smad2/3,TGF-β1 in mouse renal tissue were determined by reverse transcription-polymerase chain reaction(RT-PCR)method.Results Compared with the normal group,the FBG and UTP levels were increased significantly in the model group(P<0.05),and the obvious pathological changes were seen in renal tissue,MDA content in renal tissue was significantly increased,SOD and GSH-Px activities were decreased,ratio of p-Smad2/3 to Smad2/3,m RNA expression levels of Smad2/3 and TGF-β1 were significantly increased,and Smad7 m RNA and protein expression level was decreased significantly(all P<0.05).Compared with the model group,UTP level was decreased(P<0.05)in the two treatment groups,and the damaged renal tissue was improved,content of MDA in renal tissue was decreased,SOD and GSH-Px activities were increased,ratio of p-Smad2/3 to Smad2/3,m RNA expression levels of Smad2/3 and TGF-β1 were significantly lowered,and Smad7 m RNA and protein expression level was enhanced significantly(all P<0.05).Conclusion Salvianolate is significantly effective for improving renal function in mice with diabetic nephropathy,and the mechanism is possiblely related to the inhibition of oxidative stress and regulation of TGF-β1/Smad signaling pathway,and thus reduces renal fibrosis.
作者
杨冰
高飞
刘令今
张尧
张翠轻
檀淼
檀金川
YANG Bing;GAO Fei;LIU Ling-Jin;ZHANG Yao;ZHANG Cui-Qing;TAN-Miao;TAN Jin-Chuan(Graduate School of Hebei College of Traditional Chinese Medicine,Shijiazhuang 050000 Hebei,China;Dept.No.1 of Nephrology,the First Affiliated Hospital of Hebei College of Traditional Chinese Medicine,Shijiazhuang 050000 Hebei,China;Dept,of Endocrinology,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050000 Hebei,China)
出处
《广州中医药大学学报》
CAS
2021年第5期1018-1024,共7页
Journal of Guangzhou University of Traditional Chinese Medicine
