加味六味地黄汤对大鼠肾间质纤维化的干预机制  

Intervention Mechanism of Modified Liuwei Dihuang Tang for Renal Interstitial Fibrosis in Rats with Unilateral Ureteral Obstruction

作　　者：黄仁发 梁群卿[2] 李贺生 杨义龙 黄家晟 陈鹏辉 杨朔 吴金玉[3] HUANG Ren-Fa;LIANG Qun-Qing;LI He-Sheng;YANG Yi-Long;HUANG Jia-Sheng;CHEN Peng-Hui;YANG Shuo;WU Jin-Yu(Dept.of Nephrology,Shenzhen Hospital(Futian)of Guangzhou University of Chinese Medicine,Guangzhou 518034 Guangdong,China;Health Care Center,Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine,Nanning 530011 Guangxi,China;Dept.of Nephrology,the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023 Guangxi,China)

机构地区：[1]广州中医药大学深圳医院(福田)肾病科,广东深圳518034 [2]广西中医药大学附属瑞康医院健康体检中心,广西南宁530011 [3]广西中医药大学第一附属医院肾内科,广西南宁530023

出　　处：《广州中医药大学学报》2021年第10期2197-2202,共6页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81760805,81460682);广东省自然科学基金资助课题(编号:2020A151501572);深圳市科创委课题(编号:JCYJ201908091024131560)。

摘　　要：【目的】探讨加味六味地黄汤对大鼠肾间质纤维化的干预机制。【方法】将120只健康成年SD大鼠随机分为假手术组、模型组、中药组、缬沙坦组等4组,4组组内再按7、14、28 d 3个时间点分组,每组10只。除假手术组,其他组别大鼠均采用结扎输尿管法构建单侧输尿管梗阻模型。假手术组除不结扎输尿管外其他手术步骤均与上述造模方法相同。成功造模后,中药组术后24 h给予加味六味地黄汤2.376 g·kg^(-1)·d^(-1)灌胃,缬沙坦组术后24 h给予缬沙坦30 mg·kg^(-1)·d^(-1)灌胃,假手术组和模型组均在术后24 h给予等体积生理盐水灌胃,每日1次,直至相应实验时间点处死。采用免疫组织化学法检测肾组织c-myc、纤维连接蛋白(Fn)的表达,蛋白免疫印迹(Western Blot)法检测肾组织Wnt4、β-连环蛋白(β-catenin)、磷酸化糖原合成激酶3β(p-GSK-3β)、磷酸化T细胞因子/淋巴增强因子(p-TCF/LEF)的表达。【结果】与假手术组相同时间点比较,模型组大鼠肾组织Wnt4、β-catenin、p-GSK-3β、p-TCF/LEF、Fn和c-myc蛋白表达水平显著上调,差异均有统计学意义(P<0.01);与模型组相同时间点比较,中药组和缬沙坦组大鼠肾组织Wnt4、β-catenin、p-GSK-3β、p-LEF/TCF、Fn和c-myc蛋白的表达水平下调,差异均有统计学意义(P<0.05或P<0.01),且2个治疗组组间比较,差异无统计学意义(P>0.05)。【结论】加味六味地黄汤具有抗肾间质纤维化作用,其机制可能与阻断Wnt4/β-catenin信号通路的活化,下调Fn和c-myc蛋白的表达有关。Objective To explore the intervention mechanism of modified Liuwei Dihuang Tang for renal interstitial fibrosis in rats with unilateral ureteral obstruction(UUO).Methods One hundred and twenty healthy adult SD rats were randomly divided into 4 groups,namely sham operation group,model group,Chinese medicine group and valsartan group,the above each group was further divided into 3 sub-groups according to 3 time points including 7,14,28 days,10 rats in each group.Apart from the sham operation group,the rats of the other groups were induced into UUO model by ligation of ureter.The sham operation group was given the same other operation except for ligation of ureter.After successful modeling,the rats in Chinese medicine group were given intragastric administration of modified Liuwei Dihuang Tang of 2.376 g·kg^(-1)·d^(-1) hour 24 after operation,the valsartan group was given intragastric administration of valsartan of 30 mg·kg^(-1)·d^(-1) hour 24 after operation,and the sham operation group and model group were given intragastric administration of the same volume of normal saline hour 24 after operation,once a day,reaching at corresponding detection time points for killing.The protein expression levels of c-myc and fibronectin(Fn)in rat kidney tissue were detected by immunohistochemistry,and the protein expression levels of Wnt4,β-catenin,phosphorylated glycogen synthase kinase 3β(p-GSK-3β),phosphorylated T-cell factor/lymphoid enhancer-binding factor(p-TCF/LEF)in rat kidney tissue were detected by Western blotting assay.Results Compared with the sham operation group at the corresponding time points,the protein expression levels of Wnt4,β-catenin,p-GSK-3β,p-LEF/TCF,Fn and c-myc in rat kidney tissue were up-regulated in the model group,the difference being significant(P<0.01).Compared with the model group at the corresponding time points,the protein expression levels of Wnt4,β-catenin,p-GSK-3β,p-LEF/TCF,Fn and c-myc in rat kidney tissue were down-regulated in the Chinese medicine group and valsartan group,the

关 键 词：加味六味地黄汤 肾间质纤维化 单侧输尿管梗阻 Wnt4/β-catenin信号通路 纤维连接蛋白 c-myc 大鼠 

分 类 号：R285.5[医药卫生—中药学]