Objective To preliminarily explore the expresssion of Hedgehog signaling pathway during the regeneration of small hepatocyte-like progenitor cells (SHPCs). Methods The proliferation of SHPCs was induced by administering retrorsine combined with partial hepatectomy (R/PH) in Fisher344 rats as Gordon's method. Specific assays, such as RT-PCR, real-time PCR, immunohistochemistry and western-blot, were performed to detect the expression of the constituents related to Hedgehog signaling pathway during liver regeneration. Results RT-PCR demonstrated that the mRNA expression of IHH, PTCH, Smo, Gli1, Gli2, Gli3 were all detected in primary SHPCs cells. The results of real-time PCR analysis indicated that there were statistically significant differences in Hedgehog-related constituents (IHH, PTCH, Gli1, Gli3) in both different treatment groups and at distinct time points in the same treatment group during SHPCs regeneration (all P<0.001). The data also suggested an interaction effect existing between the treatment and the time. The protein expression of PTCH and Gli1 in liver regeneration was confirmed by immunohistochemistry, which differed in the duration in different groups. Conclusion The Hedgehog signaling pathway was continually activated during SHPCs regeneration. It's maybe an important signaling pathway during the liver regenerantion after chronic liver damage.%目的 初步探讨小肝细胞样祖细胞增殖过程中Hedgehog 信号通路的表达及其变化.方法 按Gordon's方案,Fisher344 大鼠腹腔注射倒千里光碱后,择期行部分肝切除术诱导小肝细胞样祖细胞的增殖,反转录聚合酶链反应、实时荧光定量PCR 及免疫组化等方法检测原代再生细胞均浆中Hedgehog 信号通路相关成分在不同时间点的表达,并以同期正常大鼠原代肝细胞作对照.结果 小肝细胞样祖细胞增殖过程中Hedgehog 信号通路多种成分表达,其中IHH、PTCH、Smo、Gli1、Gli2、Gli3 mRNA 持续表达并呈动态变化,SHH 表达阴性.信号分子IHH,以及反映通路活动状态的3 个标志(PTCH,Gli1,Gli3)在同一时间点与对照组之间差异有统计学意义,在实验组内不同时间点之间差异也有统计学意义(均P ＜0.001),时间与处理组之间有交互效应.免疫组化证实PTCH、Gli1 在蛋白水平的表达.结论 Hedgehog 信号通路在小肝细胞样祖细胞增殖过程中持续激活,可能是调控慢性肝损伤后肝再生的一条新的重要的信号通路.
展开▼
