补肾降浊方在糖尿病肾病中的作用及其机制研究

The role and underlying mechanism of Bushen Jiangzhuo Recipe in diabetic nephropathy

摘要:

目的 探究补肾降浊方在糖尿病肾病中的作用及其机制研究。方法 复制糖尿病肾病大鼠模型，使用11.04 g/（kg·d）补肾降浊方治疗。根据不同方法将大鼠随机分为Control组、DN组、DN + 补肾降浊方组，每组6只。通过生化分析仪测定尿样中的尿白蛋白水平、血尿素氮和血肌酐含量，HE染色检测肾组织病理损伤，Masson染色检测肾组织纤维化情况，实时荧光定量聚合酶链反应检测SIRT1 mRNA，Western blotting检测MAPK/NF-κB信号通路相关蛋白的表达。结果 DN组血尿素氮、血清肌酐、尿白蛋白水平较Control组升高（P <0.05），补肾降浊方组较DN组降低（P <0.05）。DN组SIRT1 mRNA相对表达量较Control组降低（P <0.05），DN + 补肾降浊方组较DN组升高（P <0.05）。Control组肾小球和肾小管形态完整，而DN组肾小管基底膜增厚，肾小球肥大，表明DN组大鼠肾功能受到严重损害。与DN组比较，DN + 补肾降浊方组大鼠肾小管基底膜增厚和肾小球肥大情况明显减少。与Control组比较，DN组肾小球和肾小管腔中胶原蛋白的沉积量明显增加。与DN组比较，DN +补肾降浊方组大鼠肾小球和肾小管腔中胶原蛋白的沉积量明显减少。DN组SIRT1蛋白较Control组降低（P <0.05），p-p38/p38、p-NF-κB p65/NF-κB p65较Control组升高（P <0.05）；DN +补肾降浊方组SIRT1蛋白较DN组升高（P <0.05），p-p38/p38、p-NF-κB p65/NF-κB p65较DN组降低（P <0.05）。结论 补肾降浊方促进SIRT1的表达，抑制MAPK/NF-κB信号通路，改善糖尿病肾病肾组织损伤。

Abstract:

Objective To explore the role and underlying mechanism of Bushen Jiangzhuo Recipe in diabetic nephropathy (DN).Methods A rat model of DN was established and treated with Bushen Jiangzhuo Recipe at a dose of 11.04 g/(kg?d). Based on different treatment, the rats were randomly divided into the control group, DN group, and DN + Bushen Jiangzhuo Recipe group, with 6 in each group. The levels of urinary albumin, blood urea nitrogen (BUN) and serum creatinine (Scr) were measured by biochemical analyzer. The pathological damage of renal tissues was detected by H&E staining, and the renal fibrosis was detected by Masson staining. The mRNA expression level of SIRT1 was detected by qRT-PCR, and the expression of proteins associated with the MAPK/NF-κB signaling pathway was detected by Western blotting.Results The levels of BUN, Scr and urinary albumin in the DN group were higher than those in the control group (P < 0.05), while those in the DN + Bushen Jiangzhuo Recipe group were lower compared with the DN group (P < 0.05). The relative mRNA expression of SIRT1 in the DN group was lower than that in the control group (P < 0.05), whereas that in the DN + Bushen Jiangzhuo Recipe group was higher compared with the DN group (P < 0.05). The glomeruli and tubules were morphologically intact in the control group, while thickening of the tubular basement membrane and the glomerular hypertrophy were observed in the DN group, indicating severe damage to the renal function of rats in the DN group. In comparison to the DN group, the thickening of the tubular basement membrane and the glomerular hypertrophy were mitigated in the DN + Bushen Jiangzhuo Recipe group. Compared with the control group, increased collagen deposition in glomerulus and the tubular lumen were observed in the DN group. There was a reduction in the collagen deposition in glomerulus and the tubular lumen in the DN + Bushen Jiangzhuo Recipe group compared with the DN group. The protein expression of SIRT1 in the DN group was lower than that in the control group (P < 0.05), while the levels of p-p38/p38 and p-NF-κB p65/NF-κB p65 in the DN group were higher than those in the control group (P < 0.05). The protein expression of SIRT1 in the DN + Bushen Jiangzhuo Recipe group was higher than that in the DN group (P < 0.05), while the levels of p-p38/p38 and p-NF-κB p65/NF-κB p65 in the DN + Bushen Jiangzhuo Recipe group were lower than those in the DN group (P < 0.05).Conclusion Bushen Jiangzhuo Recipe ameliorates the renal tissue damage in DN via upregulating the expression of SIRT1 and inhibiting the MAPK/NF-κB signaling pathway.