基于RUNX3/Smad3信号通路研究肾消方治疗自发性糖尿病肾脏疾病db/db小鼠肾纤维化的机制  

Study on the mechanism of Shenxiao Decoction on treating renal fibrosis in db/db mice with spontaneous diabetic kidney disease based on RUNX3/Smad3 signaling pathway

作　　者：蔡琼丽 刘文洪[2] 刘恺远 陈丽君 周迪夷[3] 周旦阳[3] 牟新[3] CAI Qiong-li;LIU Wen-hong;LIU Kai-yuan;CHEN Li-jun;ZHOU Di-yi;ZHOU Dan-yang;MOU Xin(The 2nd Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou 310053,China;Zhejiang Chinese Medical University,Hangzhou 310053,China;Endocrinology Department,Hangzhou Red Cross Hospital,Hangzhou 310003,China)

机构地区：[1]浙江中医药大学第二临床医学院,杭州310053 [2]浙江中医药大学,杭州310053 [3]杭州市红十字会医院内分泌科,杭州310003

出　　处：《中华中医药杂志》2021年第8期4573-4577,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774217,No.81273623);浙江省中医药管理局项目(No.2017ZKL016,No.2019ZB096)。

摘　　要：目的:观察肾消方对自发性糖尿病肾脏疾病(DKD)db/db小鼠肾小球系膜细胞RUNX3与肾脏纤维化的影响,探讨RUNX3与DKD的关系。方法:选取12只C57BL/KsJ-+/db雄性小鼠作为正常组,60只db/db小鼠造模成功后随机分成模型组、肾消方组、补益中气组、通利三焦组和厄贝沙坦组,每组12只。药物干预12周后全自动生化仪检测肾功能相关的参数,PAS染色观察肾脏病理变化,免疫荧光观察RUNX3在细胞中的分布,Western Blot法测定肾脏中RUNX3/Smad3信号通路关键蛋白表达情况。结果:与正常组比较,模型组小鼠24h进食量、体质量、空腹血糖、糖化血红蛋白、尿肌酐、尿素氮、尿微量白蛋白/尿肌酐、p-Smad3、H3K9me2、E-cadherin表达水平显著上升(P<0.01),而RUNX3、H3K27me3表达水平显著下降(P<0.01)。与模型组比较,肾消方组和拆方组小鼠RUNX3(补益中气组除外)、H3K27me3、尿肌酐表达水平显著上升(P<0.01,P<0.05),H3K9me2、E-cadherin、p-Smad3表达水平均显著下降(P<0.01)。结论:肾小球系膜细胞RUNX3在自发性糖尿病db/db小鼠的表达与DKD密切相关,在肾消方的干预下可以延缓糖尿病小鼠的肾脏结局。Objective:To observe the effects of Shenxiao Decoction(SXD)on glomerular mesangial cell RUNX3 and renal fibrosis in db/db mice with spontaneous diabetic kidney disease(DKD)and so as to explore between its relationship.Methods:Twelve male C57BL/KSJ-+/db mice were selected as normal group,and 60 db/db mice were randomly divided into model group,SXD group,Buyi Zhongqi group,Tongli Sanjiao group and irbesartan group after modeling,with 12 mice in each group.After 12 weeks of drug intervention,parameters related to renal function were detected by automatic biochemical analyzer,pathological changes of kidney were observed by PAS staining,the distribution of RUNX3 in cells was observed by immunofluorescence,and the expression of key proteins in the RUNX3/Smad3 signaling pathway in kidney was determined by Western Blot.Results:Compared with the normal group,24h intake,body weight,the expressions of fasting blood glucose,HbA1c,urea nitrogen,urinary creatinine,urinary microalbumin/urinary creatinine,p-Smad3,H3K9me2 and E-cadherin in model group mice at 12 weeks were significantly increased(P<0.01),the expression levels of RUNX3,H3K27me3 were significantly decreased(P<0.01).Compared with model group,the expression levels of RUNX3(the Buyi Zhongyi group exceptly),H3K27me3 and urinary creatinine in SXD and separated decoction groups were increased(P<0.01,P<0.05),while the other observation indexes(H3K9me2,E-cadherin and p-Smad3)were decreased(P<0.01).Conclusion:The expression of glomerular mesangial cell RUNX3 in spontaneous diabetic db/db mice were closely related to DKD,the intervention of SXD could delay the renal outcome of diabetic mice.

关 键 词：糖尿病肾脏疾病 肾消方 RUNX3 SMAD3 纤维化 机制 

分 类 号：R285.5[医药卫生—中药学]