组蛋白去乙酰化酶抑制剂FK228治疗小鼠肝硬化的实验研究

Experimental study of romidepsin in reducing liver cirrhosis

目的:观察组蛋白去乙酰化酶抑制剂罗米地辛(Romidepsin,FK228)对四氯化碳(carbon tetrachloride,CCl4)诱导小鼠肝纤维化模型的治疗性作用,初步探讨FK228对减轻小鼠肝纤维化的机制。方法:将30只8周龄C57BL/6雄性小鼠随机分为对照组、造模组和治疗组,每组10只;对照组腹腔注射生理盐水;造模组腹腔注射10%CCl4溶液诱导肝纤维化,持续24周;治疗组在诱导纤维化第20周开始采用FK228进行治疗。检测各组小鼠血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate transaminase,AST)含量水平;对肝组织进行HE染色和天狼星红染色,观察假小叶形态结构;进行α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)免疫组化染色,观察肝星状细胞活化程度及肝纤维化进程;Western blot半定量分析α-SMA蛋白在肝脏中的表达情况。结果:造模组血清ALT(67.270±11.109)IU/L,而治疗组小鼠的血清ALT(37.670±8.461)IU/L显著下降(P=0.019);肝组织切片HE染色和天狼星红染色显示造模组纤维化程度高、假小叶形成明显,而治疗组纤维化程度明显减弱;免疫组化染色和Western blot结果显示治疗组α-SMA水平相对于造模组明显降低(P=0.001)。结论:研究结果首次证实FK228对降低小鼠肝纤维化程度有治疗性作用,而且其机制可能与抑制星状细胞活化和降低α-SMA蛋白表达水平有关。

Objective：To observe the effect of histone deacetylase inhibitor romidepsin （FK228） on liver cirrhosis in mice, and to in-vestigate the underlying mechanism of renoprotective effect of FK228. Methods：Thirty experimental mice were randomly di- vided into three groups ： control group （group A）, model group （group B） and treatment group（group C）. To set up the liver cirrhosis model,mice in group B and C were treated with in- traperitor/eal injection of CC14 for 24 weeks. From the 21th week, group C were treated with intraperitoneal injection of FK228 weekly. The levels of serum ALT and AST were detectedto judge liver damage degree. HE and sirius red staining were used to observe pseudolobuli and tubercle. Immunohistochemical stain- ing and Western blot analysis were used to detect the expression of osteopontin α-smooth muscle actin（α-SMA） in liver. Results：The data showed the average serum ALT in group C was （37.670 ± 8.461） IU/L and （67.270 ± 11.109） IU/L in group B. It was significantly decreased when treating the experimental mice with FK228（P=-0.019）. Also, HE and sirius red staining revealed the degree of liver fi- brosis in group C were weakened obviously. Additionally, Western blot and immunohistochemistry indicated the levels of α-SMA in group C were notable reduced（P=-0.001）. Conclusion ：The preventive effect of FK228 treatment for liver cirrhosis and the mechanism may involve down regulating levels of α-SMA and inhibition of HSC activation.