超声辐照携TIMP-1 siRNA微泡对大鼠肝纤维化的影响

Effect of microbubble taking TIMP-1 siRNA to liver fibrosis of rats under ultrasound irradiation

目的探讨携TIMP-1siRNA的造影剂微泡在超声辐照条件下对肝纤维化大鼠组织学改变的影响。方法二甲基亚硝胺建立肝纤维化模型,6周后分组并静脉质粒注射,2天后观察肝、肾、肺、脑、心肌组织内基因荧光表达;12天后检测肝组织内TIMP-1mRNA、蛋白表达及胶原纤维变化。结果2天后,超声照射微泡组基因荧光表达明显增强,较其他组织差异显著,P<0.001;12天后,其TIMP-1mRNA及蛋白表达、胶原含量均较单质粒组及对照组减低,P<0.001。结论超声辐照携TIMP-1siRNA超声微泡有将目的基因定位释放作用,TIMP-1siRNA转染肝纤维化大鼠可改善肝纤维化结构,为肝纤维化的基因治疗提供了新的思路及方法。

Objective To investigate the TIMP-1 siRNA expression and the histopathological change of hepatic fibrosis by ultrasound contrast agent and ultrasound irradiation to rats liver. Methods The rat hepatic fibrosis model was established by abdominal injection of dimethylnitrosamine （DMN）, then injected TIMP-1 siRNA plasmid to penic vein at 6 weeks. One of four groups means P＋U being radiated by ultrasound in mechanical index （MI） 1.0 while injecting. Fluorescent expression of TIMP-1 siRNA was observed in ice tissues piece of liver, kendy, heart, lung and brain at 48 hours. TIMP 1 mRNA and protein expression were detected by immunohistochemistry and in situ hybridization, contain of collegen was evaluated by Masson after 12 day. Results After 2 days, fluorescent expression in liver of P＋U group was significantly higher than other tissues（P〈0. 001） ; TIMP-1 mRNA, protein and collegen expression significantly decreased in P＋U than other groups （P〈0. 001）. Conclusion Ultrasound contrast agent and ultrasound irradiation promoted gene transfection and targeting TIMP-1 siRNA to liver. TIMP-1 siRNA promoted the station of hepatic fibrosis in rats.