论文部分内容阅读
目的观察加味黄芪赤风汤对TGF-β1/Smad信号通路的影响,探讨其抗肾脏纤维化的机制。方法采用血清药理学方法,大鼠予加味黄芪赤风汤和替米沙坦灌胃,制备含药血清。体外培养小鼠系膜细胞,以脂多糖作为炎症刺激因子。实验分为6组:正常组、模型组、替米沙坦组、加味黄芪赤风汤(简称中药)高、中、低剂量组。含药血清干预72h后,采用酶联免疫法吸附试验测定各组细胞上清液中层粘连蛋白(laminin,LN)、纤维连接蛋白(fibronectin,FN)的含量;采用Western blot法检测各组细胞中TGF-β1、p-Smad2/3、Smad7、结缔组织生长因子(connective tissue growth factor,CTGF)蛋白表达。结果(1)与正常组比较,模型组上清液中LN和FN含量增加(P<0.01);与模型组比较,替米沙坦组与中药中剂量组LN及FN含量减少(P<0.05,P<0.01),中药低剂量组上清液中FN含量亦减少(P<0.01)。(2)与正常组比较,模型组TGF-β1、p-Smad2/3、CTGF蛋白表达显著增加,Smad7蛋白表达减少,差异均有统计学意义(P<0.01)。与模型组比较,替米沙坦组、中药高、中、低剂量组TGF-β1、CTGF蛋白表达均降低(P<0.01,P<0.05),替米沙坦组、中药高、中剂量组p-Smad2/3蛋白表达降低(P<0.01),中药高、中剂量组Smad7蛋白表达明显增加(P<0.05)。结论加味黄芪赤风汤能够抑制致炎因子诱导的肾小球系膜细胞分泌细胞外基质增加和肾小球系膜细胞TGF-β1/Smad信号通路的过度激活,这可能是加味黄芪赤风汤抗肾脏纤维化的机制之一。
Objective To observe the effect of Modified Astragalus and Chifeng Decoction on TGF-β1 / Smad signaling pathway and to explore its anti-renal fibrosis mechanism. Methods The method of serum pharmacology was used. The rats were treated with Modified Astragalus Chifeng Decoction and Telmisartan, and the serum containing drug was prepared. Mouse mesangial cells were cultured in vitro and lipopolysaccharide was used as an inflammatory stimulator. The experiment was divided into 6 groups: normal group, model group, telmisartan group, Modified Astragalus and Chifeng Decoction (referred to as traditional Chinese medicine) high, medium and low dose groups. Serum containing 72h after intervention, the enzyme-linked immunosorbent assay was used to determine the contents of laminin (LN) and fibronectin (FN) in the supernatant of each group. Western blot was used to detect TGF-β1, p-Smad2 / 3, Smad7, connective tissue growth factor (CTGF) protein expression. Results (1) Compared with the normal group, the content of LN and FN in the supernatant of the model group increased (P <0.01). Compared with the model group, the content of LN and FN in the medium dose group decreased , P <0.01). The content of FN in the supernatant of the traditional Chinese medicine low dose group also decreased (P <0.01). (2) Compared with the normal group, the expression of TGF-β1, p-Smad2 / 3 and CTGF in the model group increased significantly and the expression of Smad7 protein decreased in the model group (P <0.01). Compared with the model group, the expression of TGF-β1 and CTGF in the telmisartan group and the traditional Chinese medicine group were significantly decreased (P <0.01, P <0.05). The telmisartan group, The protein expression of p-Smad2 / 3 decreased (P <0.01), and the expression of Smad7 in high-dose and medium-dose groups of traditional Chinese medicine increased significantly (P <0.05). Conclusion Modified Astragalus and Chifeng Decoction can inhibit the increase of extracellular matrix secretion induced by inflammatory cytokines and the over-activation of TGF-β1 / Smad signaling pathway in glomerular mesangial cells, which may be flavored astragaluschiforme soup Anti-renal fibrosis mechanism.