红景天甙对肝纤维化大鼠肝组织ROCK表达的影响

Effects of salidroside on expression of ROCK in rats with liver fibrosis

目的:研究红景天甙对肝纤维化大鼠肝组织ROCK基因表达的影响,了解其干预肝纤维化的机制.方法:健康雄性SD大鼠,随机分为3组:对照组(n=10),红景天甙干预组(n=40)和肝纤维化模型组(n=40).大鼠肝纤维化采用CCl4皮下注射法(300mL/L,3mL/kg,每周2次,共8wk)诱导.红景天甙干预采用腹腔注射法,剂量5mg/kg,每周2次.肝脏胶原沉积状况采用Masson胶原染色以及HE染色观察,ROCKⅠ、ROCKⅡ表达水平分别采用原位杂交(in situ hybridization,ISH)和免疫组化(im munohistochemistry,IH)方法检测.实验图像经电子计算机扫描,数据采用专业图像分析软件统计分析.结果:肝纤维化大鼠肝脏肝细胞坏死、再生明显,假小叶形成,胶原纤维沉积明显增加,肝实质结构紊乱.红景天甙干预组大鼠肝组织胶原沉积明显减少,组织学积分平均胶原面积降低(2.1±0.3vs3.6±0.8,74.82±21.51μm2vs290.86±89.37μm2,均P<0.05).与模型组比较,红景天甙干预性治疗组ROCKⅠ,Ⅱ蛋白表达水平均明显下降(0.203±0.068vs0.357±0.182,0.237±0.056vs0.394±0.238,均P<0.05),ROCKⅠ,ⅡmRNA表达水平明显下降(0.197±0.019vs0.394±0.238,0.185±0.031vs0.279±0.112,均P<0.01).结论:红景天甙可能通过调节Rho-ROCK信号传导通路,减少肝脏胶原纤维的合成与沉积,有效干预CCl4诱导的大鼠肝纤维化.

AIM： To observe the effects of salidroside on the expression of ROCK in liver tissue of CCl4- induced liver fibrosis in rats, and to explore its probable mechanism. METHODS： Ninety healthy SD rats were ran- domly divided into 3 groups： control group （n = 10）, salidroside group （n = 40） and liver fibrosis group （n = 40）. Experimental liver fibrosis was induced by （with the concentration of 300 mL/L soluted in liquid paraffin） subcutaneous injection of CC14 （at the dosage of 3 mL/kg, twice per wk, 8 wks）. The salidroside was injected into the peritoneal cavity at the dosage of 5 mg/kg, twice per week for 8 weeks. Liver tissues from each group were stained with Masson and HE staining to observe the collagen deposition. Expressions of ROCK I and ROCK Ⅱ in the liver tissue were detected with in situ hybridization （ISH） and immunohistochemistry （IH） respectively. All the figures were scanned with electronic computer, and the data were analyzed with Image-Plus software. RESULTS： A significant collagen deposition and rearrangement of the parenchyma were noted in liver tissue of CCl4-treated rats. There were lots of pseudolobule in liver tissue. The semiquantitative histological scores and average area of collagen were significantly increased when compared with control rats （2.1±0.3 vs 3.6 ± 0.8, 74.82± 21.51 μm^2 vs 290.86±89.37μm^2, both P 〈 0.05）. Compared with control group, the expres- sions of ROCK I, ROCK Ⅱ and ROCK I mRNA, ROCK Ⅱ mRNA were decreased significantly in salidroside group （0.203 ± 0.068 vs 0.357 ± 0.182, 0.237 ± 0.056 vs 0.394 ± 0.238; 0.197 ± 0.019 vs 0.394 ± 0.238, 0.185 ± 0.031 vs 0.279 ± 0.112, P 〈 0.05 or 0.01）. CONCLUSION： The expressions of ROCK I and ROCK Ⅱ in liver tissues are inhibited significantly with salidroside treatment. Salidroside could interfere with the signal transduction of Rho- ROCK pathway and then inhibit liver fibrosis in rats.