口服肝靶向制剂介导受体ASBT和ASGPR在肝泡型包虫病大鼠模型体内的表达分析  

Expression of ASBT and ASGPR mediated receptors for oral liver-targeting preparations in a rat model of hepatic alveolar echinococcosis

作　　者：高瑞雪 胡春晖[1] 张发斌[1] 高攀 甘雪辉 张耀刚 姜博璠 GAO Ruixue;HU Chunhui;ZHANG Fabin;GAO Pan;GAN Xuehui;ZHANG Yaogang;JIANG Bofan(Medical College of Qinghai University,Xining 810001,China;Qinghai Province Key Laboratory of Hydatid Disease Research,Xining 810001,China)

机构地区：[1]青海大学医学院,西宁810001 [2]青海省包虫病研究重点实验室,西宁8100011.8伦理学审查2.2ASBT在HAE大鼠模型回肠组织中的表达与分布

出　　处：《临床肝胆病杂志》2021年第4期846-851,共6页Journal of Clinical Hepatology

基　　金：国家自然科学基金项目(81960641);青海大学省部共建三江源生态与高原农牧业国家重点实验室自主课题项目(2019-ZZ-01)。

摘　　要：目的通过测定顶端钠依赖性胆盐转运体(ASBT)和去唾液酸糖蛋白受体(ASGPR)的表达研究,探讨其在肝泡型包虫病(HAE)药物治疗方面口服肝靶向制剂设计的可行性。方法18只雄性SD大鼠,10只大鼠造模,造模成功的8只大鼠为HAE组,正常组大鼠8只作为对照。采用免疫荧光、Western Blot、实时荧光定量PCR对ASBT在HAE模型大鼠及正常大鼠的回肠组织的表达分布、蛋白及基因表达水平进行检测分析;采用同样的检测方法分析ASGPR在HAE大鼠模型的非患病肝组织、肝组织病灶边缘带及正常大鼠肝组织的表达水平。符合正态分布的计量资料2组间比较采用t检验,3组间比较采用单因素方差分析,两两比较采用LSD-t检验。结果免疫荧光、Western Blot、实时荧光定量PCR结果均显示ASBT在HAE组回肠组织的表达较正常组表达上调(t值分别为5.309、4.110、28.060,P值均<0.05)。免疫荧光、Western Blot、实时荧光定量PCR结果均显示,ASGPR在正常对照组大鼠肝组织、HAE组大鼠非患病肝组织和HAE组大鼠肝织病灶边缘带中表达差异有统计学意义(F值分别为110.666、128.201、143.879,P值均<0.001),其中HAE组较正常组表达水平高,且越靠近病灶组织表达越高。结论ASBT和ASGPR可作为HAE潜在的口服肝靶向制剂的介导受体,为设计用于治疗HAE的口服肝靶向制剂提供理论基础。Objective To investigate the feasibility of apical sodium-dependent bile salt transporter(ASBT)and asialoglycoprotein receptor(ASGPR)in the design of oral liver-targeting preparations for the treatment of hepatic alveolar echinococcosis(HAE)by measuring the expression of ASBT and ASGPR.Methods A total of 18 male Sprague-Dawley rats were selected,among which 10 were used to establish a model of HAE(HAE group)and 8 were used as controls(normal group).Immunofluorescence assay,Western blotting,and quantitative real-time PCR were used to measure the expression distribution,protein expression level,and mRNA expression level of ASBT in the ileal tissue of HAE model rats and normal rats;the same methods were used to measure the expression level of ASGPR in the non-diseased liver tissue and the marginal zone of liver tissue lesion of HAE model rats and the liver tissue of normal rats.The t-test was used for comparison of normally distributed continuous data between two groups;a one-way analysis of variance was used for comparison between three groups,and the least significant difference t-test was used for comparison between two groups.Results The results of immunofluorescence assay,Western blotting,and quantitative real-time PCR showed that compared with the normal group,the HAE group had significantly upregulated expression of ASBT in the ileal tissue(t=5.309,4.110,and 28.060,all P<0.05)and a significantly higher expression level of ASGPR(the closer to the lesion,the higher the expression)(F=110.666,128.201,and 143.879,all P<0.001).Conclusion ASBT and ASGPR can be used as potential mediated receptors for oral liver-targeting preparations for HAE,which provides a theoretical basis for the design of oral liver-targeting preparations for the treatment of HAE.

关 键 词：棘球蚴病 肝 分子靶向治疗 顶端钠依赖性胆盐转运体 去唾液酸糖蛋白受体 大鼠 Sprague-Dawley 

分 类 号：R532.32[医药卫生—内科学]