论文部分内容阅读
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。
OBJECTIVE: To observe the appropriate dose and effect of hepatic regeneration stimulating factor (HSS) and epidermal growth factor (EGF) on the proliferation and regeneration of hepatocytes and to speculate the phase of HSS. Methods: The proliferation of cells was detected by ~ 3HTdR incorporation method. HSS were added into primary rat hepatocytes cultured in vitro for different time. The cell growth was detected by MTT assay. RESULTS AND CONCLUSIONS: 1.SHSS (≤100μg / ml) and EGF (≤100ng / ml) had significant stimulatory effects on hepatic derived cells (P <0.01), and there was a dose-dependent relationship between them. However, . There was a synergistic effect between HSS and EGF. After cultured in primary hepatocytes and hepatoma cell lines (SMMC-7721, BEL-7402) for 48 h, the cpm values of (~ 3H) TdR incorporation assay were 5.94 , 2.98 and 3.36 times. In primary cultured rat hepatocytes, the stimulating effect of adding HSS after adhering to Oh.20h was more significant than that of 40h, suggesting that HSS may play a major role in the G1 / S phase of hepatocyte regeneration.