肝再生刺激因子、表皮生长因子对肝细胞的作用

Effect of HSS and EGF on Hepatocytes

目的：观察肝再生刺激因子(HSS）及表皮生长因子（EGF)对肝细胞增殖再生作用合适剂量及其联合效应，并推测HSS作用时相。方法：以~3HTdR掺入法检测细胞DNA增殖；体外原代大鼠肝细胞培养不同时间加入HSS,MTT法检测细胞生长状况。结果与结论：1.HSS（≤100μg／ml)和EGF（≤100ng／ml）对肝衍生细胞均有显著刺激作用（P＜0.01）,并存在剂量关系，但剂量过大，刺激作用反而下降。2.HSS、EGF存在协同作用，原代肝细胞、肝癌细胞株（SMMC-7721、BEL-7402）体外培养48h后，经(~3H）TdR掺入法检测cpm值分别为各自对照组的5.94、2.98和3.36倍。3.体外培养原代大鼠肝细胞，于贴壁后Oh.20h加入HSS其刺激作用较40h组显著.提示HSS可能主要作用于肝细胞再生G1/S期。

Background／Aims:To determine the proper dosage and the combination effect of HSS and EGF on DNA synthesis of hepatocytes in culture and their acting phase.Methods:Studied the effect of different dosage of HSS and EGF on DNA synthesis of rat hepatocytes in primary culture and hepatoma cell lines(SMM-7721,BEL-7402)with the assay of^3H TdR incorporation,also deduced HSS function on cell cycle with MTT assay.Results:1.HSS (≤100μg／ml),EGF(≤100ng／ml)stimulated DNA synthesis of hepatocytes in primary culture and hepatoma cell line with dose dependent manner,but less effect with higher dose.2.HSS, EGF acted synergistically,the^3 H TdR incorporation of hepatocytes,SMMC-7721 and BEL-7402 were increased to 5.94,2.98 and 3.36-fold respectively after 48h of culture,compared with controls.3.DNA synthesis of rat hepatocytes in primary culture where the time of HSS addition was varied and the magnitude of Oh and 20h groups were much higher,Conclusions:It is suggested that HSS might be acted on G1／S phase mostly.