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电针联合两色金鸡菊提取物对糖尿病肾病大鼠肾脏Vimentin、α-SMA、TGF-β1及p-smad2表达的影响

电针联合两色金鸡菊提取物对糖尿病肾病大鼠肾脏Vimentin、α-SMA、TGF-β1及p-smad2表达的影响

蒋洁 阿米拉?阿布拉提 姚蓝

摘要:目的  观察电针联合两色金鸡菊提取物对糖尿病大鼠肾脏波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)及p-smad2表达的影响,探讨其作用机制。方法  将36只SD大鼠随机分为正常组、模型组、电针组、两色金鸡菊组、电针+两色金鸡菊组和二甲双胍组,每组6只。采用高糖高脂喂养联合腹腔注射链脲佐菌素建立2型糖尿病模型。正常组、模型组不进行干预;电针组针刺大鼠双侧“肾俞”“脾俞”,连接电针,留针30 min;两色金鸡菊组予两色金鸡菊乙酸乙酯提取物(300 mg/kg)灌胃;电针+两色金鸡菊组予电针联合两色金鸡菊乙酸乙酯提取物灌胃;二甲双胍组予二甲双胍(200 mg/kg)药液灌胃。每日1次,治疗4周。HE染色观察肾组织形态学及纤维化程度,Western blot检测肾组织Vimentin、α-SMA、TGF-β1及p-smad2蛋白表達。结果  HE染色结果显示,电针组、两色金鸡菊组、电针+两色金鸡菊组和二甲双胍组均可缓解模型大鼠肾小管扩张,减轻肾小球皱缩现象,改善炎性细胞浸润,减少基底膜增生和融合,且以电针+两色金鸡菊组改善更明显;Western blot检测结果显示,电针和两色金鸡菊提取物均可降低模型大鼠肾组织Vimentin、α-SMA、TGF-β1及p-smad2蛋白表达,且以两者联合效果最佳。结论  电针联合两色金鸡菊提取物可明显改善糖尿病大鼠肾脏纤维化,其作用机制可能与抑制TGF-β1/Smad2信号通路有关。

关键词:电针;两色金鸡菊提取物;糖尿病肾病;波形蛋白;α-平滑肌肌动蛋白;转化生长因子-β1;Smad2;大鼠

中图分类号:R245;285.5    文献标识码:A    文章编号:1005-5304(2019)06-0055-04

Abstract: Objective To observe the effects of electroacupuncture combined with coreopsis tinctoria extract for expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 in renal tissue of diabetic rats; To discuss its mechanism of action. Methods Totally thirty-six SD rats were randomly divided into normal group, model group, electroacupuncture group, coreopsis tinctoria group, electroacupuncture combined with coreopsis tinctoria group and metformin group, with 6 rats in each group. Type 2 DM model was established by high-fat and high-glucose diet and STZ injection intraperitoneally. Normal group and model group were given no treatment; Electroacupuncture was at the acupoints of “Pishu” and “Shenshu” once daily in electroacupuncture group for 30 min; coreopsis tinctoria group was given intragastric administration of 300 mg/kg coreopsis tinctoria extract for gavage; electroacupuncture combined with coreopsis tinctoria group was given electroacupuncture and intragastric administration of coreopsis tinctoria extract for gavage; metformin group was given intragastric administration of 200 mg/kg metformin for gavage. Histological changes and fibrosis degree of kidneys were observed by HE staining and expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 were detected by Western blot. Results HE staining results indicated that

糖尿病肾病(diabetic nephropathy,DN)已成为终末期肾病的第2位致病因素[1]。终末期肾病一旦形成,治疗极为棘手。因此,及时治疗DN,防止其向终末期肾病转化意义重大。肾脏纤维化是DN主要的病理特征之一,前期研究发现,两色金鸡菊提取物对高糖高脂诱导的大鼠肾小球系膜纤维化有明显抑制作用[2-3],而电针也可改善DN肾功能和超微结构[4]。本实验拟将电针联合两色金鸡菊提取物干预DN大鼠,观察其对DN大鼠肾脏波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)及p-smad2表达的影响,明确两者是否存在协同作用,为临床治疗DN提供依据。

1  材料与方法

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