摘要
目的探讨超声辐照携载短发夹RNA干扰增殖1阻断(block of proliferation1,BOP1)基因的超声微泡造影剂治疗大鼠肝纤维化的可行性。方法成功构建大鼠肝纤维化模型,将40只肝纤维化大鼠随机分为携载质粒的超声微泡组(BOP1-shRNA+US/MB)、携载质粒的常规超声组(BOP1-shRNA+US)、单纯质粒组(BOP1-shRNA)及空白对照组(CON),每组10只。BOP1-shRNA+US/MB组及BOP1-shRNA+US组分别经股静脉注入1mL微泡溶液及1mL携载单纯质粒的生理盐水后,行肝区超声辐照。所有分组大鼠在干预21d后行磁共振扩散加权成像检查后麻醉脱颈处死,取肝脏组织行苏木精-伊红(HE)染色观察病理变化。应用Western blot法检测BOP1蛋白在大鼠肝脏组织中的表达,检验BOP1的沉默效果。结果 BOP1-shRNA+US/MB组的表观弥散系数(ADC)值高于其他3组,而指数表观弥散系数(EADC)值低于其他3个组(P<0.05)。HE染色示BOP1-shRNA+US/MB组肝脏细胞排列尚整齐,汇管区纤维组织增生,见少量炎性细胞浸润,小叶结构完整,其肝纤维化程度低于其他组。Western blot结果显示BOP1-shRNA+US/MB组大鼠肝脏组织的BOP1蛋白表达敲减成功。结论超声辐照携载BOP1基因短发夹RNA的微泡造影剂可成功治疗大鼠肝纤维化,可为肝纤维化的精准靶向治疗提供新的理论依据。
Objective To evaluate the feasibility of employing ultrasound-targeted microbubble destruction which mediated BOP1 gene silencing to treat rat live fibrosis. Methods Rat models of liver fibrosis were successfully constructed. Rats were divided into four groups：BOPI-shRNA＋US/MB,BOPI-shRNA＋US, simple plasmid group （BOPI-shRNA） and blank control group （CON）,with 10 rats in each group. Croup BOPI-shRNA＋US/MB and BOPI-shRNA＋US were injected with lmL microbubble solution via femoral vein and lmL normal saline with plasmid. After intervention for 21d, all rats were sacrificed by magnetic resonance diffusion weighted imaging （DWI）, and liver tissue was stained with Hematoxylin Eosin （HE）. Western blot was used to detect the expression of BOP1 protein in rat liver, and to test the silencing effect of BOP1. Results The apparent diffusion coefficient （ADC） of group BOPI-shRNA＋US/MB was higher than that of the other 3 groups, while the index apparent diffusion coefficient （EADC） value was lower than that of the other 3 groups （P〈0.05）. HE staining showed that the liver cells in group BOPI-shRNA＋US/MB were neatly arranged, fibrous tissue proliferated in the pipe area,a small amount of inflammatory cells were infiltrated, and the structure of the lobule was complete, and the degree of liver fibrosis was lower than that of the other groups. Western blot results showed that the expression of BOP1 protein in liver tissue of BOPI-shRNA＋US/MB group was knocked down successfully. Conclusion The microbubble contrast agent carrying BOP1 gene short hairpin RNA can successfully treat the liver fibrosis in rats. It can provide a new theoretical basis for the precise targeting therapy of liver fibrosis.
作者
马丽园
姜韬
姜艺
王乐
杨光飞
米成嵘
MA Liyuan;JIANG Tao;JIANG Yi;WANG Le;YANG Guangfei;MI Chengrong(Department of Ultrasound,the General Hospital of Ningxia Medical University,Yinchuan 750004;Department of Anal-Colorectal Surgery,the General Hospital of Ningxia Medical University,Yinchuan 750004)
出处
《宁夏医科大学学报》
2018年第6期641-643,653,共4页
Journal of Ningxia Medical University
基金
国家自然科学基金(81560394)
宁夏自然科学基金(NZ15161)
关键词
超声联合微泡造影技术
BOP1基因
肝纤维化
基因治疗
大鼠
microbubble contrast agent combined ultrasound techniques
block of proliferationl
liver fibrosis
gene therapy
rat
