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目的探讨内皮素-1(ET-1)致系膜细胞细胞外基质(ECM)合成增加的介导途径。方法体外培养人肾小球系膜细胞,按培养条件不同分为正常对照组、ET-1组、阻断剂(RG50864)干预组及氯沙坦干预组,分别培养24 h。采用ELISA法测定细胞上清液中IV型胶原(Col IV)、纤维连接蛋白(Fn)的含量;RT-PCR法测定培养系膜细胞内血小板源性生长因子-B(PDGF-B)mRNA的表达,Western blot法检测PDGF-B蛋白的表达。结果①在外源性ET-1作用下,上清液中Col IV、Fn蛋白含量、系膜细胞PDGF-B mRNA及蛋白表达增加(P<0.05),以10-7mol/L ET-1作用最为明显。RG50864呈浓度依赖方式抑制ET-1所致的Col IV、Fn分泌增加(P<0.05);②氯沙坦能抑制ET-1所致系膜细胞PDGF-B mRNA及蛋白表达增加,减少上清液中Col IV、Fn蛋白的含量(P<0.05)。结论①ET-1能够刺激ECM分泌增加并促进系膜细胞合成PDGF-B,ET-1所致的系膜细胞ECM分泌增加可能部分由PDGF-B介导;②氯沙坦可以通过拮抗系膜细胞PDGF-B的产生进而减少ECM的分泌和沉积,从而产生明显的肾脏保护作用。
Objective To investigate the mediation of the increased synthesis of extracellular matrix (ECM) in mesangial cells induced by endothelin-1 (ET-1). Methods Human glomerular mesangial cells were cultured in vitro and divided into normal control group, ET-1 group, RG50864 intervention group and Losartan intervention group according to different culture conditions, and cultured for 24 hours respectively. The contents of type IV collagen (Col IV) and fibronectin (Fn) in the cell supernatant were measured by ELISA. The expression of platelet-derived growth factor-B (PDGF-B) The expression of PDGF-B protein was detected by Western blot. Results ① Under the action of exogenous ET-1, the concentration of Col IV, Fn and the expression of PDGF-B mRNA and protein in mesangial cells increased (P <0.05), and the effect of ET-1 at 10-7 mol / L obvious. RG50864 inhibited the secretion of Col IV and Fn induced by ET-1 in a concentration-dependent manner (P <0.05); ② Losartan inhibited the expression of PDGF-B mRNA and protein in mesangial cells induced by ET-1, The contents of Col IV and Fn in fluid (P <0.05). Conclusions ①ET-1 can stimulate the secretion of ECM and promote the synthesis of PDGF-B by mesangial cells. The increase of ECM secretion induced by ET-1 may be partially mediated by PDGF-B; ② Losartan can antagonize the proliferation of mesangial cells The production of PDGF-B, in turn, reduces the secretion and deposition of ECM resulting in significant renal protection.