携带hIL-10基因的腺病毒对大鼠肝纤维化的实验研究

Experimental research about the effect of adenovirus-mediated hIL-10 gene on liver cirrhosis of mice

目的探讨在体内试验中携带人白介素(hIL)-10基因的腺病毒是否有较高的转染效率、能够发挥抗炎和抗纤维化的作用以及hIL-10表达时间的长短。方法向四氯化碳(CCl4)诱导的肝脏纤维化模型大鼠尾静脉中注射含携带hIL-10基因的腺病毒载体的林格液,观察其与不经过CCl4处理的肝脏纤维化模型大鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、肝纤维化4项指标(血清透明质酸、层黏连蛋白、Ⅲ型前胶原、Ⅳ型胶原)以及病理结果对比;以增强绿色荧光蛋白(EGFP)作为荧光标记,观察腺病毒的转染效率;并使用ELISA、Western blot检测hIL-10在大鼠体内的表达情况。结果携带hIL-10基因的腺病毒有较高的转染效率;在大鼠体内,能够发挥抗炎和抗纤维化的作用;在血浆中,8h hIL-10的表达量达到最高,之后逐渐下降,1周之内代谢完全。结论携带hIL-10基因的腺病毒对肝病的治疗可能有广阔的应用前景。

Objective To explore whether adenovirus that carries hIL-10 gene has high transduction efficiency, an- ti-inflammatory and anti-fibrosis function as well as to test expression time of hIL-10, in vivo. Methods CC14 was used to induce mice models with liver cirrhosis. Then, Ringer＇s solution containing adenovirus -mediated IL-10 was injected into mice with liver cirrhosis through caudal vein. Later, ALT, AST and four index signs （ HA, LN, PC m, iv-c） of mice models were observed, and compared with those without injection. At the same time, adenovirus^s transduction efficiency was observed by using fluorescence microscope for the fluorescence of green fluorescent pro- tein. And expression time of hIL-10 was observed in vivo by using ELISA and Western blot methods. Results Ade- novirus-mediated IL-10 gene has high transfection efficiency, and has the function of anti-inflammatory and anti-fi- brosis in vivo. In blood plasma, hIL-10 expression level reached highest in 8 hours, and then decreased. After one week,hIL-10 totally disappeared. Conclusion Adenovirus with hIL-10 gene may have a promising future in trea- ting liver disease.