阿魏酸哌嗪对TGF-β_1诱导下肾小球系膜细胞结缔组织生长因子表达及细胞外基质合成的影响

Effects of Piperazine Ferulate on Connective Tissue Growth Factor and Extracellular Matrix in TGF-β_1 Induced Mesangial Cells

目的探讨阿魏酸哌嗪(PF)对转化生长因子-β1(TGF-β1)诱导下肾小球系膜细胞结缔组织生长因子(CTGF)及细胞外基质成分表达的影响及其意义。方法将体外培养正常大鼠肾小球系膜细胞(HBZY-1)分为正常对照组、TGF-β1刺激组、TGF-β1加50、100、200ng/mLPF干预组。分别利用免疫细胞化学法检测各组细胞CTGF蛋白表达,实时荧光定量逆转录-聚合酶链反应(RT-PCR)测定细胞CTGFmRNA表达量,双抗体夹心ABC-ELISA法检测细胞上清型胶原(Col)、纤维连接蛋白(FN)的含量。结果CTGF蛋白及mRNA、细胞上清Col、FN表达量在TGF-β1组较正常组明显升高(P<0.05),阿魏酸哌嗪组上述指标均低于TGF-β1组(P<0.05)。结论阿魏酸哌嗪能抑制TGF-β1诱导下肾小球细胞外基质(ECM)的合成。其机制可能与下调系膜细胞上CTGF的表达量及功能活性有关。

Objective To observe the effects of piperazine ferulate （PF） on synthesis of extracellular matrix and expression of connective tissue growth factor （CTGF） in transforming growth factor-betal （TGF-β1） induced rat glomerular mesangial cells （GMC）, and to investigate its mechanism on the prevention of glomerulosclerosis. Methods Rat HBZY-1 GMCs were cultured in vitro and divided into 5 groups： Normal control group, TGF-β1-wstimulated group, TGF-β1 plus 50, 100, 200 ng/mL PF group. The level of CTGF protein expression induced by TGF-β1 was measured by immunocytochemistry, The mRNA expression of CTGF was evaluated by quantitative real time fluroscent polymerase chain reaction. The expressions of fibronectin and collagen I were analyzed by enzyme linked immunosorbent assay （ELISA）. Results With the stimulation of TGF-β1, the expression of of CTGF protein and mRNA, as well as fibronectin and collagen I were markly increased（P〈0.05）, which then were decreased by the treatment of PF. （P〈0. 05）. Conclusion PF can partly inhibit synthesis of the extracellular matrix and CTGF. PF may take part in the prevention of glomerulosclerosis through the inhibition of CTGF expression.