文献类型：期刊文献

中文题名：超声作用下CTGF-siRNA对大鼠肝纤维化基因治疗的实验研究

英文题名：The Experimental Study of CTGF-siRNA Gene Therapy for Liver Fibrosis in Rats Under Ultrasound

作者：程祖胜[1];夏国园[2];钱彩艳[2];董梁[2];孙建刚[2];江珍珍[3];王爱芝[3]

机构：[1]绍兴市第七人民医院,浙江省绍兴市312000;[2]绍兴文理学院附属医院,浙江省绍兴市312000;[3]绍兴市人民医院,浙江省绍兴市312000

年份：2021

卷号：37

期号：1

起止页码：93

中文期刊名：中国超声医学杂志

外文期刊名：Chinese Journal of Ultrasound in Medicine

收录：CSTPCD、、CSCD2021_2022、北大核心、CSCD、北大核心2020

基金：浙江省自然科学基金项目(No.Y2110966);浙江省医药卫生研究计划A类项目(No.2015KYA225、2013KYB269、2014KYA217)。

语种：中文

中文关键词：肝纤维化;超声;CTGF-siRNA;基因治疗

外文关键词：Hepatic fibrosis;Ultrasound;CTGF-siRNA;Gene therapy

中文摘要：目的探讨利用超声微泡造影剂介导携带结缔组织生长因子-小干扰RNA(CTGF-siRNA)真核表达质粒转染大鼠肝细胞的有效性。方法 (1)构建CTGF-siRNA真核表达质粒;(2)将40只实验大鼠随机分为7组:正常组,实验对照组,CTGF-siRNA质粒组,超声联合微泡组,超声联合微泡介导基因低、中、高剂量组;(3)建立肝纤维化模型,基因治疗4周后处死大鼠,超声及病理切片HE染色评价肝纤维化程度,masson染色观察胶原纤维含量,反转录-聚合酶链反应(RT-PCR)检测肝组织中CTGF、Ⅰ型和Ⅲ型胶原的mRNA表达。结果 (1)CTGF-siRNA基因治疗后超声及病理切片显示干预组纤维化程度降低;(2)masson染色显示超声介导微泡基因组胶原纤维含量表达量随着剂量增高而降低(P<0.05);(3)RT-PCR显示超声介导微泡基因组CTGF、Ⅰ型和Ⅲ型胶原的mRNA表达明显低于其他组,且随着质粒剂量增大而减少(P<0.05)。结论超声微泡作用下CTGF-siRNA基因能特异地作用于靶位点,有效抑制肝纤维化进程,提高对肝纤维化干预的特异度。

外文摘要：Objective To investigate the efficiency of the transfection of connective tissue growth factor-small interfering RNA(CTGF-siRNA) eukaryotic expression plasmid mediated by ultrasound microbubbles to transfect rat hepatocytes. Methods(1)Construction of eukaryotic expression plasmid of CTGF-siRNA.(2)40 experimental rats were randomly divided into 7 groups. The normal group, the experimental control group,the CTGF-siRNA plasmid group, ltrasound+microbubbles group, ultrasound+microbubbles+low,medium,high-dose plasmid group.(3)Establishment of liver fibrosis model.Rats were killed after 4 weeks of gene therapy. Ultrasound and pathological section HE staining was used to evaluate the degree of liver fibrosis. Masson tricolor staining was used to observe the content of collagen fibers. RT-PCR was used to detect the mRNA expression of CTGF, I and type Ⅲ collagen in liver tissue. Results(1)Ultrasound and pathological examination showed that the intervention group reduced the degree of fibrosis after CTGF-siRNA gene therapy.(2)Masson tricolor staining showed that the content of collagen fibrils in the ultrasound mediated microbubble genome decreased with the increase of dose(P<0.05).(3)The expression of mRNA in CTGF,Ⅰ and type Ⅲ collagen in each group of microbubbles mediated by reverse transcription-polymerase chain reaction(RT-PCR)was significantly lower than that in other groups,and with the increase of the dose of plasmid decreased(P<0.05). Conclusions Under the action of ultrasound microbubbles, the CTGF-siRNA gene can play a more special role in the target loci. It effectively inhibits the process of liver fibrosis and improves the specificity of liver fibrosis intervention.