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目的观察蟾蜍灵对血小板源性生长因子(plate derived growth factor-BB,PDGF-BB)诱导的系膜细胞(me-sangial cell,MC)PAI-1、u-PA、t-PA的表达影响,探讨其对系膜细胞纤溶酶原/纤溶酶系统的作用。方法 PDGF(20ng/ml)刺激体外培养的系膜细胞12h后,加入0.08μM蟾蜍灵干预12h,用ELISA法检测细胞上清培养液中PAI-1的表达量;RT-PCR法测定PAI-1、u-PA、t-PA基因表达变化。结果 PDGF-BB可诱导MC中PAI-1的mRNA表达上调,并且表达增加,u-PA、t-PAmRNA转录下调;0.08μM蟾蜍灵可下调PDGF诱导的系膜细胞的PAI-1mRNA,上调u-PA、t-PAmR-NA的转录,并抑制PAI-1的表达。0.08μM蟾蜍灵对正常系膜细胞无明显影响(P>0.05)。结论蟾蜍灵可通过促进细胞外基质降解来抑制PDGF-BB诱导的肾小球硬化。
Objective To observe the effect of bufalin on the expression of PAI-1, u-PA and t-PA induced by platelet derived growth factor-BB (PDGF-BB) in mesangial cells (MCs) To investigate its effect on mesangial cell plasminogen / plasmin system. Methods The mesangial cells cultured in vitro were stimulated with PDGF (20ng / ml) for 12h, and 0.08μM of bufalin was added to the mesangial cells for 12h. The expression of PAI-1 in the culture supernatant of the cells was detected by ELISA. PAI-1 , U-PA, t-PA gene expression changes. Results PDGF-BB could up-regulate the mRNA expression of PAI-1 in MC and increase the expression of u-PA and t-PA mRNA. In addition, 0.08μM Bufalin down-regulated the PAI-1 mRNA in mesangial cells induced by PDGF, PA, t-PAmR-NA transcription and inhibition of PAI-1 expression. 0.08μM toad on the normal mesangial cells had no significant effect (P> 0.05). Conclusion Bufalin can inhibit PDGF-BB-induced glomerulosclerosis by promoting extracellular matrix degradation.