Du Yue-Guang, Zhang Ke-Na, Gao Zong-Lei, Dai Fengjiao, Wu Xi-Xi, Chai Ke-Fu
Department of Pathology, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, P.R. China.
National Traditional Chinese Medicine Clinical Research Center, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, P.R. China.
Exp Ther Med. 2018 Feb;15(2):2156-2164. doi: 10.3892/etm.2017.5621. Epub 2017 Dec 12.
The present study investigated the mechanism underlying the anti-inflammatory effects of Tangshen formula (TS) in Sprague Dawley (SD) rats with diabetic nephropathy (DN). A rat model of DN was established by intraperitoneal injection of 1% (40 mg/kg) streptozotocin and administration of a high fat and glucose diet. Subsequently, SD rats were randomly divided into six groups (n=8): A DN group, a valsartan group, a high-dose TS group, a middle-dose TS group, a low-dose TS group and a control group with normal SD rats. Once rats received their allocated treatment for 12 weeks, body weight and kidney weight were recorded, and fasting blood glucose, ratio of urinary protein, β-MG and creatinine clearance rate were determined. Furthermore, hemodynamic indices, including plasma viscosity and whole blood reduction viscosity were detected. Immunohistochemistry was used to detect the infiltration of macrophages in the kidneys of rats. Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to investigate the activation; mRNA and protein expression levels of monocyte chemoattractant protein-1 (MCP-1), macrophage migration inhibitory factor (MIF), nuclear factor-κB (NF-κB) and sirtuin-1 (SIRT1) in each group. In comparison with the DN group, each biochemical indicator of rats in the high-dose TS group was significantly decreased (P<0.05). Blood viscosity in each treatment group was significantly decreased when compared with the DN group (P<0.01). Hematoxylin and eosin staining indicated that the infiltration of macrophages was significantly decreased in the high-dose TS group when compared with the DN group (P<0.01). mRNA and protein expression levels of MCP-1 and MIF in the high-dose TS group were significantly decreased when compared with the DN group (P<0.05). In the treatment groups, SITR1 mRNA expression levels were significantly increased, whereas the mRNA expression levels of NF-κB were significantly decreased (P<0.01). Western blotting results indicated a significant decrease in the protein expression levels of acetylated NF-κB in the treatment groups when compared with the DN group (P<0.01) and the propensity of protein expression of the other inflammatory factors were consistent with the mRNA findings. The results of the high-dose TS group were similar to those of the valsartan group. The present study indicates that TS was able to activate SITR1, which lead to NF-κB deacetylation, thus reducing the release of inflammatory factors and decreasing the severity of diabetic nephropathy.
本研究探讨了糖肾方（TS）对糖尿病肾病（DN）Sprague Dawley（SD）大鼠抗炎作用的潜在机制。通过腹腔注射1%（40 mg/kg）链脲佐菌素并给予高脂高糖饮食建立DN大鼠模型。随后，将SD大鼠随机分为六组（n = 8）：DN组、缬沙坦组、高剂量TS组、中剂量TS组、低剂量TS组和正常SD大鼠对照组。大鼠接受分配的治疗12周后，记录体重和肾脏重量，并测定空腹血糖、尿蛋白比值、β-微球蛋白和肌酐清除率。此外，检测血流动力学指标，包括血浆黏度和全血还原黏度。采用免疫组织化学法检测大鼠肾脏中巨噬细胞的浸润情况。进行逆转录-定量聚合酶链反应和蛋白质印迹法以研究各组中单核细胞趋化蛋白-1（MCP-1）、巨噬细胞移动抑制因子（MIF）、核因子-κB（NF-κB）和沉默信息调节因子1（SIRT1）的激活、mRNA和蛋白表达水平。与DN组相比，高剂量TS组大鼠的各项生化指标均显著降低（P < 0.05）。与DN组相比，各治疗组的血液黏度均显著降低（P < 0.01）。苏木精-伊红染色显示，与DN组相比，高剂量TS组巨噬细胞浸润显著减少（P < 0.01）。与DN组相比，高剂量TS组MCP-1和MIF的mRNA和蛋白表达水平显著降低（P < 0.05）。在治疗组中，SITR1 mRNA表达水平显著升高，而NF-κB的mRNA表达水平显著降低（P < 0.01）。蛋白质印迹结果表明，与DN组相比，治疗组中乙酰化NF-κB的蛋白表达水平显著降低（P < 0.01），其他炎症因子的蛋白表达倾向与mRNA结果一致。高剂量TS组的结果与缬沙坦组相似。本研究表明，TS能够激活SITR1，导致NF-κB去乙酰化，从而减少炎症因子的释放并降低糖尿病肾病的严重程度。
