肾元颗粒影响糖尿病小鼠肾脏中miR-199b-5p与钙磷代谢的研究

Research about Effect of Shenyuan Granules on miR-199b-5p and Calcium-phosphorus Metabolism in Diabetic Nephropathy Mice

目的:探讨肾元颗粒对糖尿病小鼠肾脏中miR-199b-5p/Klotho/FGFR-1/EGR-1通路及钙磷代谢的影响。方法:链脲佐菌素(Streptozotocin,STZ)诱导小鼠糖尿病模型,模型成功后随机分为肾元颗粒组[4.55 g/(kg·d)]10只和模型对照组10只,另设空白对照组10只。给予相应药物灌胃8周,给药前后检测血糖以及血清中肌酐(SCr)、尿素氮(BUN)、钙(Ca)和磷(P),采用RT-q PCR检测肾脏组织中miR-199b-5p、Klotho、FGFR-1和EGR-1核糖核酸表达情况,采用western-blot(WB)法检测Klotho、FGFR-1和EGR-1蛋白表达情况,检测小鼠右下肢平均骨密度。结果:用药后与空白对照组相比,模型对照组P、SCr升高(P<0.05),Ca、骨密度明显降低(P<0.05),同时miR-199b-5p和EGR-1表达上调(P<0.05),Klotho和FGFR-1表达下调(P<0.05)。用药后与模型对照组比较,肾元颗粒组P、SCr降低(P<0.05),Ca升高(P<0.05),miR-199b-5p、EGR-1表达下调(P<0.05),Klotho、FGFR-1表达上调(P<0.05)。结论:肾元颗粒缓解糖尿病肾损害和改善钙磷代谢异常的作用机制,或与miR-199b-5p/Klotho/FGFR-1/EGR-1通路有关。

Objective： To explore the effect of Shenyuan Granules on miR - 199b -5p/Klotho/FGFR - 1/EGR - 1 signaling pathway and calcium- phosphorus metabolism in diabetic nephropathy（DN） mice model. Method： STZ- induced DN mice models were randomly divided into Shenyuan Granules group[4.55 g/（kg ·d） ] （n = 10） and model control group （ n = 10） ,with the blank control group （ n = 10） established independently. Corresponding drugs were given by gavage for 8 weeks. Before and after treatment, blood glucose, blood urea nitrogen （ BUN）, serum creatinine （ SCr）, calcium （Ca） ,phosphor （P） and bone density were detected. Moreover, the RT -qPCR was used to detect the mRNAs of miR - 199b - 5p, Klotho, FGFR - 1 and EGR - 1. The Western - blot method was used to detect the protein of Klotho, FGFR - 1 and EGR - 1. Result： Compared with the blank control group, serum P and SCr levels of mice in model control group were significantly increased （ P 〈 0.05 ）, serum Ca and bone density levels were significantly decreased （ P 〈 0.05 ）, expressions of miR -199b -5p and EGR -1 were up -regulated（P 〈0.05）. And the expressions of Klotho and FGFR -1 were down - regulated （ P 〈 0.05 ）. Compared with the blank control group, serum P and SCr levels of mice in Shenyuan Granules group were significantly decreased （P 〈 0.05 ）, serum Ca were increased （ P 〈 0. 05 ）, expressions of miR - 199b -5p and EGR -1 were down -regulated（ P 〈 0.05 ）. And the expressions of Klotho and FGFR -1 were upregulated （ P 〈 0. 05 ）. Conclusion ： Shenyuan Granules improving the renal function and calcium - phosphorus metabolism indexes in diabetic nephropathy mice models may be connected with miR - 199b - 5p/Klotho/FGFR - 1/EGR - 1 signaling pathway.