Caveolin-1 在肝纤维化中作用的研究

摘要:目的 通过 10%四氯化碳( CCl₄ )橄榄油溶液,分别诱导野生型小鼠和小窝蛋白-1( Caveolin-1) 敲除小鼠建立肝纤维化模型,比较分析敲除 Caveolin-1 对肝损伤和胶原沉积的影响,揭示其在肝纤维化中的作用。 方法 按剂量 1 μL / g,腹腔注射野生型小鼠和 Caveolin-1 敲除小鼠 10% CCl₄ 橄榄油溶液,2 次 / 周。 注射后,1、3、7、14 和 28 d,分 别处死 5 只小鼠,取血清和肝脏,通过苏木素-伊红( HE) 染色、血清谷丙转氨酶( ALT) 和谷草转氨酶( AST) 水平检测,分析肝脏损伤程度;利用天狼猩红染色、Real-time PCR 及 Western blot 方法,观察肝脏胶原沉积及肝纤维化标志物 α 平滑肌肌动蛋白( α-SMA) 、Ⅰ 型胶原蛋白( collagen-Ⅰ )和Ⅲ 型胶原蛋白( collagen-Ⅲ ) mRNA 及蛋白表达水平, 分析肝纤维化程度。 结果 与野生型小鼠相比,造模后 3 d,敲除小鼠的血清 ALT 和 AST 含量显著升高( P<0. 01) ;3、7 和 14 d,敲除小鼠肝脏坏死面积显著增大( P<0. 01 或 P<0. 05) ;7、14 及 28 d 时,敲除小鼠胶原染色面积明显增多( P<0. 01 或 P<0. 05) ;肝星状细胞活化标志物 α-SMA mRNA 水平在 3 d 和 14 d 时显著升高(P<0. 05),collagen-Ⅰ和collagen-Ⅲ mRNA 在 14 d 时显著上调(P<0. 01);14 d 时,敲除小鼠肝脏 α-SMA、collagen-Ⅰ及 collagen-Ⅲ 蛋白表达水平均显著升高( P<0. 01 或 P<0. 05) 。 结论 Caveolin-1 抑制 CCl₄ 诱导的小鼠肝脏炎性损伤及纤维化。

Study on the Role of Caveolin-1 in Liver Fibrosis

Abstract: Objective In this study, wild-type and Caveolin-1 ( Caveolin-1) knockout mice were induced by 10%carbon tetrachloride ( CCl₄) olive oil solution to establish liver fibrosis model. The effect of Caveolin-1 knockout on liver injury and collagen deposition was analyzed to explore the role of Caveolin-1 in liver fibrosis. Method Wildtype mice and Caveolin-1 knockout mice were intraperitoneally injected with 10% CCl₄ olive oil solution at 1 μL / g body weight, twice a week. Five mice were sacrificed on day 1, 3, 7, 14 and 28 after the injection. Serum and liver were collected, and the degree of liver injury was analyzed by HE staining and serum ALT and AST levels. The liver collagen deposition, together with the mRNA and protein expression of α-SMA, CollagenⅠ- and Collagen-Ⅲ were observed by Sirius red staining, Real-time PCR and Western blot to analyze the degree of liver fibrosis. Result On day 3, the contents of ALT and AST in serum of the knockout mice were significantly increased( P<0. 01) . On day 3, 7 and 14, the necrotic area of liver of knockout mice was significantly increased ( P<0. 01 or P<0. 05) . On day 7, 14 and 28, the collagen staining area in Caveoin-1 knockout mice increased significantly ( P<0. 01 or P<0. 05) . Hepatic stellate cell activation marker, α-SMA mRNA significantly increased on day 3 and day 14 ( P<0. 05) . mRNA expression of Collagen- Ⅰ and Collagen-Ⅲ rise significantly on day 14 ( P <0. 01) . The protein expression levels of α-SMA, Collagen- Ⅰ and Collagen-Ⅲ were significantly higher in Caveolin-1 knockout mice ( P<0. 01 or P<0. 05) . Conclusion Caveolin-1 inhibited the liver inflammatory injury and fibrosis in mice induced by CCl₄.