Rho相关卷曲螺旋形成蛋白激酶1在大鼠肝纤维化不同阶段表达及意义

Rho - associated coiled - coil forming protein kinase 1 expression in different stages of hepatic fibro- sis

目的观察Rho相关卷曲螺旋形成蛋白激酶1（ROCK1）在大鼠肝纤维化过程中不同阶段的表达。方法72只200—250g雄性Wistar大鼠随机分为正常对照组36只、肝纤维化模型组36只。正常对照组即正常饲养，模型组即腹腔注射四氯化碳，2次／周，剂量2ml／kg。分别在3、6d、2、4、6、8周时间点随机选取6只大鼠处死，苏木素-伊红（HE）染色观察肝脏组织的病理学变化，观察肝纤维化程度。免疫组织化学方法检测ROCK1的组织定位及表达。利用实时定量聚合酶链反应（Real—timePCR）方法检测不同时间段ROCK1基因mRNA转录水平的变化。结果免疫组织化学结果显示，给药后3、6d、2、4、6、8周，正常对照组ROCK1阳性表达率分别为11％、15％、13％、11％、17％、14％；模型组ROCK1阳性表达率分别为14％、15％、52％、65％、75％、80％；差异有统计学意义（P=0．001）。Real—timePCR检测显示ROCK1表达量（以对照组为1），模型组表达分别为1．018759、1．381913、1．831238、2．515323、2．257623、2．188587。结论ROCK1在四氯化碳诱导大鼠肝纤维化的过程中，其组织表达水平逐渐升高，说明Rho／ROCK通路可能是肝纤维化发生的另一条重要信号通路。

Objective To investigate the expression of Rho -associated coiled -coil forming pro- tein kinase 1 （ ROCK1 ） in different stages of hepatic fibrosis. Methods Seventy - two Wistar male rats of 200 to 250 g were selected and randomly divided into control group and model group. The control group was fed normally, and the model group was given 2 ml/kg of carbon tetrachloride （ CC14 ） intraperitoneally, twice a week. The animals （ n = 6） were killed at 3rd, and 6th day and 2nd, 4th, 6th, and 8th week. He- matoxylin and eosin （HE） staining was used to observe the pathological changes of live tissues. Immuno- histochemistry was used to detect the location and expression of ROCK1. ROCK1 gene transcripts in different periods were quantitatively detected by real - time quantitative polymerase chain reaction （ Real - time PCR）. Results The positive expression rate of ROCK1 in control group was 11% , 15% , 13% , 11% , 17% and 14%, and that in model group was 14%, 15%, 52%, 65%, 75% and 80% at 3rd, and 6th day and 2nd, 4th, 6th, and 8th week, respectively, with significant differences among them （P = 0. 001 ）. Realtime PCR revealed that the ROCK1 mRNA expression was 1.018 759, 1.381 913,1.831 238, 2. 515 323,2. 257 623,2. 188 587 in model group at 3rd, and 6th day and 2nd, 4th, 6th, and 8th week when that in control group was corrected to 1. Conclusion The expression level of ROCK1 protein increased gradually in the formation and development of liver fibrosis, suggesting the Rho/ROCK signaling pathway may be another important signaling pathway of liver fibrosis.