左归降糖益肾方对糖尿病肾病MKR鼠肾组织VDR、CYP27B1表达的影响

Effects of Zuogui Jiangtang Yishen Decoction on the expressions of renal VDR, CYP27B1 of MKR mice with diabetic nephropathy

目的:探讨左归降糖益肾方对糖尿病肾病(DN)MKR鼠肾组织VDR、CYP27B1表达的影响。方法:8周龄MKR鼠随机分为假手术组、模型组(高脂喂养联合单侧肾切除)、中药(左归降糖益肾方)组、阳性药物组(格列喹酮片与盐酸贝那普利片),每组15只;正常组选取同龄C57鼠10只。造模8周后灌胃给药,治疗8周。留取尿标本,心脏采血处死各组小鼠。电化学法检测空腹血糖,常规生化法检测尿蛋白含量;取新鲜肾组织,实时荧光定量PCR检测肾组织VDR、CYP27B1 mRNA的表达水平;Western blot法检测肾组织VDR、CYP27B1蛋白的表达水平;HE及电镜观察肾组织病理变化。结果:与模型组比较,中药组能升高DN小鼠体质量(P<0.05),降低肾质量、肾肥大指数、空腹血糖、尿蛋白含量、血清肌酐、尿素氮含量(P<0.01);上调VDR、CYP27B1的mRNA及蛋白的表达水平(P<0.01);改善肾组织病理损伤结构。结论:左归降糖益肾方可以通过调控VDR、CYP27B1的表达水平,改善肾组织病理结构损伤。

Objective:To observe the effects of the Zuogui Jiangtang Yishen Decoction(ZJYD)on the expressions of renal VDR and CYP27 B1 of MKR mice with diabetic nephropathy(DN).Methods:Eight weeks of MKR mice were randomly divided into sham operation group,model group(high-fat diet combined with unilateral nephrectomy),ZJYD group(Zuogui Jiangtang Yishen Decoction)and positive drug group(Gliclazin Tablets and Benazemate Hydrochloride),15 mice each group;the normal group was selected from 10 C57 mice of the same age.Eight weeks after model establishment,the mice were gavaged for eight weeks.The urine samples were taken and the mice were sacrificed by cardiac blood sampling.The levels of FBG was observed by electrochemical method;the levels of urine protein content were observed by routine biochemical method;and the expressions of VDR and CYP27 B1 mRNA in renal tissue was detected by real-time fluorescent quantitative PCR.The expression of VDR and CYP27 B1 protein in renal tissue was detected by Western blot.HE and electron microscope were performed to assess pathological changes in the renal tissues.Results:Compared with the model group,the ZJYD group could increase body weight of DN mice(P<0.05);decrease renal mass,renal hypertrophy index,fasting blood glucose,urinary protein content,serum creatinine,and urea nitrogen of DN mice(P<0.01);up-regulate the mRNA and protein expressions of VDR and CYP27 B1(P<0.01);improve the pathological lesion of renal tissue.Conclusion:ZJYD can inhibit the development of renal injure by upregulating the expressions of VDR and CYP27 B1.