糖肾宁对糖尿病性肾脏病小鼠足细胞Nephrin、Synaptopodin蛋白表达的影响  

Effects of Tangshenning on expression of nephrin and synaptopodin of podocyte in diabetic kidney disease mice

作　　者：崔方强 王悦芬[1] 江心灿 赵文景[1] CUI Fang-qiang;WANG Yue-fen;JIANG Xin-can;ZHAO Wen-jing(Beijing Hospital of Traditional Chinese Medicine Affiliated to the Capital Medical University,Beijing 100010,China;School of Traditional Chinese Medicine Affiliated to the Capital Medical University,Beijing 100069)

机构地区：[1]首都医科大学附属北京中医医院肾病科,北京100010 [2]首都医科大学中医药学院,北京100069

出　　处：《北京中医药》2020年第10期1028-1032,F0003,共6页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81703989);北京市自然科学基金项目(7182069,7172096)。

摘　　要：目的观察糖肾宁对糖尿病性肾脏病(DKD)小鼠足细胞功能蛋白Nephrin及Synaptopodin的作用。方法 8周龄KK-Ay小鼠高脂饲料喂养4周建立DKD小鼠模型后,随机分为模型组、糖肾宁组和缬沙坦组,另选取相同月龄的C57BL/6J小鼠作为正常对照组。糖肾宁组予20 g/(kg·d)的糖肾宁(生药)灌胃,缬沙坦组予10 mg/(kg·d)缬沙坦灌胃,正常对照组及模型组予等剂量蒸馏水灌胃,连续灌胃12周。在灌胃0、4、8、12周后收集尿液检测24 h尿蛋白定量,灌胃12周后心尖取血检测血肌酐及尿素氮水平,摘取肾脏,HE、PAS、Masson染色观察肾脏病理,免疫组化检测足细胞Nephrin、Synaptopodin蛋白表达,RT-PCR检测足细胞Nephrin、Synaptopodin的mRNA表达。结果与正常对照组比较,模型组小鼠24 h尿蛋白定量、血肌酐、血尿素氮水平明显升高(P<0.05);与模型组比较,糖肾宁组及缬沙坦组小鼠24 h尿蛋白、血肌酐、血尿素氮水平明显降低(P<0.05)。与正常对照组比较,模型组小鼠肾小球系膜细胞明显增多,细胞外基质明显增生;与模型组比较,糖肾宁组及缬沙坦组小鼠的肾小球系膜细胞明显减少,细胞外基质增生明显减轻。与正常对照组比较,模型组小鼠足细胞Nephrin、Synaptopodin蛋白及mRNA水平显著下调(P<0.05);与模型组比较,糖肾宁组及缬沙坦组小鼠足细胞Nephrin、Synaptopodin蛋白及mRNA水平显著上调(P<0.05)。结论糖肾宁可以降低DKD小鼠尿蛋白水平,改善肾功能,机制可能与其上调足细胞Nephrin、Synaptopodin蛋白表达,减轻足细胞损伤有关。Objective To observe the effects of Tangshenning(TSN) on nephrin and synaptopodin of podocyte in diabetic kidney disease(DKD) mice.Methods The DKD mice model was established by feeding 8-week old KK-Ay mice with high-fat diet for 4 weeks and then randomly divided into model group,TSN group and Valsartan group.One more normal control group(NC) was the same age of C57 BL/6 J mice.TSN group were given TSN(raw herbs) with 20 g/(kg·d)to the stomach.Valsartan group was given Valsartan to the stomach with 10 mg/(kg·d). NC group and model group were given equal amount of saline to the stomach.All mice were successively administrated for 12 weeks.After 0,4,8,and 12 weeks,urine was collected and urine protein was measured for 24 hours.After 12 weeks,blood was taken from the apex to measure the levels of serum creatinine and urea nitrogen.Kidney was removed for the ovbservation of pathological changes by HE,PAS,and Masson staining.The protein expression of nephrin and synaptopodin in podocytes was detected by immunohistochemistry,and mRNA expression of nephrin and synaptopodin in podocytes was detected by RT PCR.Results Compared with NC group,24 h proteinuria,serum creatinine and blood urea nitrogen was significantly increased(P<0.05) in model group.Compared with model group,24 h proteinuria,serum creatinine and blood urea nitrogen was significantly decreased(P<0.05) in TSN group and Valsartan group.Compared with NC group,the glomerular mesangial cells of model group were obvious increased,extracellular matrix obviously proliferated.Compared with the model group,the glomerular mesangial cells in TSN group and Valsartan group were obvious decreased,extracellular matrix obviously was alleviated.Compared with NC group,nephrin and synaptopodin of podocyte and mRNA level were significantly decreased in the model group(P<0.05);Compared with the model group,nephrin and synaptopodin of podocyte and mRNA level were significantly increased in TSN group and Valsartan group(P<0.05).Conclusion TSN can reduce proteinuria and ameliorate

关 键 词：糖肾宁 糖尿病性肾脏病 足细胞 Nephrin蛋白 Synaptopodin蛋白 小鼠 

分 类 号：R285.5[医药卫生—中药学]