黄芪对TGF-β1致人腹膜间皮细胞分泌细胞外基质的影响

目的 :探讨黄芪对腹膜纤维化的作用及可能机制。方法 :人腹膜间皮细胞 (HPMC)作体外培养 ,在第三代 ,将细胞转板至细胞融合后 ,将其分为对照组 (F12 ) ,TGF β1组 (F12 +TGF β15ng/ml) ,TGF β1加黄芪 1组(F12加TGF β1加黄芪 ,黄芪终浓度为 10 0 μg/ml,TGF β1加黄芪 2组 (F12加TGF β1加黄芪 ,黄芪终浓度为 1mg/ml) ,分别在 2 4,48h采用ELISA法检测上清液中纤维连接蛋白 (FN)和纤溶酶原抑制剂 (PAI 1)的含量 ;FNmRNA ,PAI 1mRNA和碱性成纤维细胞生长因子 (bFGF)mRNA的表达采用半定量逆转录多聚酶链反应 (RT PCR)检测。结果 :①HPMC表达FN ,PAI 1和bFGF ;②HPMC在 5ng/mlTGF β1刺激下分泌FN及PAI 1明显的增加 ,与对照组比较差异有显著性意义 (P <0 .0 5 ) ;③HPMC在TGF β1和不同浓度的黄芪刺激后 ,在 2 4h内可使FN及PAI 1减少 ,尤其是PAI 1与TGF β1组比差异有显著性意义 (P <0 .0 5 )。④HPMC经TGF β1作用 12h后 ,FNmRNA ,PAI 1mRNA和bFGFmRNA半定量结果与对照组相比 ,其表达量分别上调 18.5 4%,8.6 %和 6 6 .9%,HPMC经 10 0μg/ml和 1mg/ml黄芪作用 12h后FN ,PAI 1和bFGFmRNA的表达与TGF β1组比较均下调 ,其结果分别为5 .3 %,1.3%,3 .8%和 5 .4%,74.3 %,2 3 .7%。结论 :TGF β1可促进HPMC合成

Influence of huangqi on TGF-beta 1 inducing extracellular matrix secretion of human peritoneal mesothelial cell

Objective To study the effect of huangqi on peritoneal sclerosis and its possible mechanism.Methods Isolated human peritoneal mesothelial cells (HPMC) was cultured . Cultured HPMC were treated with F12 without serum (control group), F12 with TGF-beta1(TGF-beta1 group),F12 with TGF-beta1 and huangqi 100 μg/ml (huangqi 1 group), F12 with TGF-beta1 and huangqi 1 mg/ml (huangqi 2 group).Fibronectin(FN) and plasminogen activator inhibitor-1(PAI-1)were detected by ELISA method. The expression of FN mRNA,PAI-1 mRNA and bFGF mRNA was detected by RT-PCR method. Results ①HPMC expressed FN,PAI-1,and bFGF. ②Fn and PAI-1 significantly increased compared with control with 5 ng/ml TGF-beta1stimulated at 24 hours and 48 hours(P<0.05); ③The different concentration of huangqi decreased FN and PAI-1 expression compared with TGF-beta1 group,especially 1 mg/ml huangqi at 24 hours(P<0.05). ④FN,PAI-1and bFGF mRNA expression were higher in 12hours after stimulation with TGF-beta1 compared with control.FN,PAI-1and bFGF mRNA expression were lower in 12 hours after stimulation with different concentration of huangqi (100 μg/ml and 1mg/ml) than those in the TGF-beta1 group.Conclusion TGF-beta1 promotes extracellular matrix synthesis and secretion of HPMC. Huangqi partly counteracts the synthesis of human peritoneal mesothelial cell’s extracellular matrix by the stimulation of TGF-beta1.