大鼠2-乙酰胺基芴/部分肝切除模型中卵圆细胞增殖动力学的初步研究

目的 研究大鼠肝脏卵圆细胞增殖模型中卵圆细胞增殖率的变化,进一步了解卵圆细胞生物学特性.方法 采用2-乙酰胺基芴/部分肝切除(2-AAF/PH)的方法建立大鼠肝卵圆细胞增殖模型.免疫荧光双标技术检测肝切除术后不同时间点肝脏石蜡切片中卵圆细胞标志物上皮细胞黏附分子和增殖标志物增殖细胞核抗原的共表达情况,对阳性细胞进行定量计数分析;并采用荧光双标和天狼猩红染色观察此过程中肝星状细胞激活和肝脏胶原沉积的情况.结果 术后第2天门静脉区开始出现少量卵圆细胞,到第9天数量达到高峰,此后数量逐步下降.此过程中卵圆细胞的增殖率逐步下降,从第2天的(91.3±1.6)%下降到第12天的(53.6±4.4)%,差异有统计学意义(P＜0.01).激活的肝星状细胞一直伴随卵圆细胞增殖并向肝实质延伸,分泌的胶原形成细胞外基质包绕小管样结构的卵圆细胞.结论 2-AAF/PH模型中卵圆细胞数量达到高峰前,其增殖率已经开始逐渐下降.肝星状细胞可能通过分泌细胞外基质和多种因子严格调控卵圆细胞的增殖.

A study of oval cell proliferation kinetics in the rat 2-acethlaminofluorene/partial hepatectomy model

Objective To investigate the changes of oval cell proliferation rate in the rat 2-acethlaminofluorene/partial hepatectomy(2-AAF/PH)model.Methods Livers were collected from 2-AAF/PH rats at different time points after hepatectomy.Paraffin sections were investigated by double immunofluorescent staining with confocal microscopy for oval cell marker epithelial cell adhesion molecule and proliferative index proliferating cell nuclear antigen,or epithelial cell adhesion molecule and alphasmooth muscle actin Deposition of matrix in liver tissue was detected by sirius red staining.Results Response of ductular oval cells could be observed in portal area at 2 days after PH,and the number of oval cells reached its peak at 9 days and then gradually declined.Oval cell proliferation rate decreased from (91.3±1.6)% at 2 days after PH to(53.6±4.4)% at 12 days(P＜0.01).In addition,oval cells infiltrating into liver parenchyma were closely associated with activated hepatic stellate cells and extracellular matrix.Conclusions Oval cell proliferation rate starts decreasing before its number reaches a peak in 2-AAF/PH model Hepatic stellate cells probably tightly regulate oval cell number through secreting several factors and producing extracellular matrix.