肝切除患者血清对体外培养肝细胞增殖能力的影响

Effects of serum came from hepatectomized patient on proliferation of cultured hepatocytes

目的通过体外细胞实验观察肝切除患者术前及术后不同时间血清对肝细胞增殖的影响。方法根据所采用的血清不同,将培养的HL-7702细胞分为胎牛血清(FBS)组、术前血清组、术后0.5h、3h、24h和72h血清组,各组细胞在Cell-IQ无标记活细胞影像分析系统中维持培养72h,通过该系统对细胞进行连续计数,绘制出各组细胞扩增曲线,比较各组细胞72h内细胞扩增倍数,对各组培养72h后的细胞进行BrdU免疫荧光染色,比较各组BrdU阳性率。结果细胞扩增曲线显示各组细胞均有增殖,除术后72h血清组外,其余人血清组培养的HL-7702细胞增殖速度均高于FBS组,术后0.5h和术后3h血清组更明显。除术后72h血清组外,各组人血清组HL-7702细胞扩增倍数均高于FBS组,术后0.5h和术后3h血清组扩增倍数高于术前血清组(均P<0.05)。人血清组HL-7702细胞BrdU阳性率均高于FBS组,术后0.5h和术后3h血清组细胞BrdU阳性率高于术前血清组(均P<0.05)。结论肝切除术后外周血清较FBS具有更强的促进肝细胞增殖的能力,肝切除术后早期血清促进肝细胞增殖作用更明显。

Objective To observe the influence of peripheral serum came from patients with hepatectomy at different time points on hepatocyte proliferation in vitro. Methods According to the different types of cultured serum, cultured HL-7702 cells were divided into fetal bovine serum (FBS) group, preoperative serum group, 0.5 h, 3 h, 24 h and 72 h post operative serum groups. All groups of cells were cultured for 72 hours in the Cell-IQ unmarked living cell image analysis system, and the amplification curves of each group were mapped by continuous counting of cells. The cell amplification multiple was compared between all groups after culturing for 72 hours. BrdU immunofluorescence staining was performed and BrdU positive rate was calculated for comparing the cell proliferation of all groups. Results Amplification curves showed that HL-7702 cell proliferation rates of all human serum groups except for 72 h post operative group were higher than those of FBS group. Human serum 0.5 h and 3 h postoperative groups were more obvious. The amplification multiples of human serum groups, except for 72 h post operative group were all significantly higher than those of FBS group (P < 0.01),and 0.5 h and 3 h post operative groups were both significantly higher than those of preoperative group (P < 0.05). BrdU positive rates of all human serum groups were significantly higher than those of FBS group (P < 0.01), which were significantly higher in 0.5 h and 3 h post operative groups than those of preoperative group (P < 0.05), but there were no statistical differences between 24 h and 72 h post operative groups and the preoperative group. Conclusion Human serum can promote the proliferation of hepatocytes compared with that of FBS. The influence of serum acquired post hepatectomy is closely associated with the post operative time.