PCI-neo-肝细胞生长因子对脂多糖刺激的大鼠肾小球系膜细胞α-平滑肌肌动蛋白表达的影响

摘要

目的：探讨 PCI-neo-肝细胞生长因子（PCI-neo-HGF）对脂多糖（LPS）刺激的大鼠肾小球系膜细胞（GMCs）的α-平滑肌肌动蛋白（α-SMA）的表达是否有抑制作用，以及这种抑制作用是否与对转化生长因子-β1（TGF-β1）表达的抑制有关。方法体外培养 GMCs 并分为5组，（1）Ctrl：对照；（2）C＋L：GMCs＋ LPS（10μg／ml）；（3）C＋L＋P-n：GMCs＋LPS（10μg／ml）＋ PCI-neo（2μg）；（4）C＋L＋P-n-H2：GMCs＋LPS（10μg／ml）＋PCI-neo-HGF（2μg）；（5）C＋L＋P-n-H8：GMCs＋ LPS（10μg／ml）＋PCI-neo-HGF（8μg）；用 RT-PCR 的方法测定α-SMA 及β-actin mRNA 的表达，用 Western 方法检测及α-SMA，TGF-β1及β-actin 的蛋白表达。结果与 Ctrl 组相比，C＋L 及 C＋L＋P-n 组α-SMA mRNA 及蛋白表达增多（P ＜0．05），TGF-β1蛋白表达增多（P ＜0．05）；与 C＋L 相比，C＋L＋P-n-H2及 C＋L＋P-n-H8组α-SMA mRNA 及蛋白的表达均减少（P ＜0．05），TGF-β1蛋白的表达减少（P ＜0．05）。与 C＋L组比较，加入 TGF-β1抑制剂组α-SMA 蛋白表达水平下降（P ＜0．05）。结论 PCI-neo-HGF 可抑制 LPS 刺激的GMCs 的α-SMA 的 mRNA 及蛋白表达，且这种抑制作用可能与其对 TGF-β1表达的抑制有关。%Objective To inquire whether PCI-neo-hepatocyte growth factor (HGF)could inhabit the express ofα-smooth muscle actin (α-SMA)in rat glomenrulus mesangial cells (GMCs)stimulated by lipopolysaccharide (LPS),and whether this effect was associated with transforming growth factor-β1 (TGF-β1).Methods GMCs were divided into the following groups:Ctrl:control;C+L:GMCs+ LPS(10 μg/ml);C+L+P-n:GMCs+LPS(10 μg/ml)+PCI-neo(2μg);C+L+P-n-H2:GMCs+LPS(10 μg/ml)+PCI-neo-HGF(2 μg);C+L+P-n-H8:GMCs+LPS(10 μg/ml)+PCI-neo-HGF(8 μg).α-SMA andβ-actin mRNA expressions were detected by RT-PCR.The protein expressions ofα-SMA,β-actin and TGF-β1 were detected by Western.Results Compared with Ctrl group,the mRNA and protein expresses ofα-SMA increased (P <0.05),and the protein express of TGF-β1 increased(P <0.05)in C+L group and C+L+P-n group.Compared with C+L group,the mRNA and protein expresses ofα-SMA increased (P <0.05),and the protein express of TGF-β1 increased(P < 0.05)in C+ L+ P-n-H2 group and C+ L+ P-n-H8 group.Compared with C+ L group,the express ofα-SMA protein decreased in adding inhibition of TGF-β1 group(P <0.05).Conclusion PCI-neo-HGF could inhibit mRNA and protein expresses ofα-SMA in rat GMCs stimulated by LPS,and this effect maybe was associated with inhibiting TGF-β1.