發(fā)布時(shí)間：2018-03-03 21:55

  本文選題：丙型肝炎　切入點(diǎn)：RNA干擾　出處：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2017年20期 　論文類(lèi)型：期刊論文

【摘要】：目的研究攜帶針對(duì)丙型肝炎病毒(hepatitis C virus,HCV)RNA的小干擾RNA(siRNA)的維生素E(Vitamin E)脂質(zhì)體納米顆粒在體外靶向抑制HCV復(fù)制的作用。方法采取"薄膜水化法"制備Vitamin E脂質(zhì)體納米顆粒,應(yīng)用激光粒度分析儀測(cè)量納米顆粒VE-DC/siRNA的顆粒大小及zeta電位,應(yīng)用透射電子顯微鏡觀(guān)察VE-DC形態(tài)及分散性。以Huh7.5.1細(xì)胞株作為載體。CCK-8法檢測(cè)納米顆粒的細(xì)胞毒性,瓊脂糖凝膠電泳檢測(cè)納米顆粒攜帶siRNA的血清穩(wěn)定性,蛋白印跡技術(shù)和免疫熒光技術(shù)檢測(cè)該siRNA納米顆粒的轉(zhuǎn)染效率及對(duì)HCV的抑制作用。結(jié)果 Vitamin E脂質(zhì)體納米顆粒形態(tài)呈球形,粒度分布為(125.5±9.0)nm,zeta電位為(39.1±4.8)m V,分散均一。相較同濃度商品化脂質(zhì)體,VE-DC/siRNA對(duì)細(xì)胞的毒性相對(duì)較小。血清穩(wěn)定性實(shí)驗(yàn)顯示,VE-DC/siRNA在24 h后siRNA無(wú)降解,較裸siRNA穩(wěn)定性顯著提高。攜帶siRNA的Vitamin E脂質(zhì)體納米顆粒可以較高效率進(jìn)入Huh7.5.1細(xì)胞內(nèi),并抑制HCV核心蛋白的表達(dá)。結(jié)論 Vitamin E脂質(zhì)體納米顆�？梢赞D(zhuǎn)運(yùn)siRNA至Huh7.5.1細(xì)胞,并抑制HCV核心蛋白的表達(dá)。
[Abstract]:Objective to study the inhibition of HCV replication by liposome nanoparticles carrying small interfering RNAs against hepatitis C virus (HCV) virus. Methods Vitamin E liposome nanoparticles were prepared by "membrane hydration method". The particle size and zeta potential of VE-DC/siRNA nanoparticles were measured by laser particle size analyzer, and the morphology and dispersion of VE-DC were observed by transmission electron microscope. The cytotoxicity of nanoparticles was detected by using Huh7.5.1 cell line as carrier. CCK-8 method was used to detect the cytotoxicity of nanoparticles. Agarose gel electrophoresis was used to detect the serum stability of the nanoparticles carrying siRNA, Western blot and immunofluorescence techniques were used to detect the transfection efficiency of the nanoparticles and their inhibitory effect on HCV. Results the Vitamin E liposome nanoparticles were spherical in shape. The particle size distribution was 125.5 鹵9.0nmnmdeta potential (39.1 鹵4.8mV). The cytotoxicity of VE-DCR siRNA was relatively less than that of commercial liposome at the same concentration. Serum stability test showed that VE-DCR / siRNA did not degrade after 24 h. Vitamin E liposome nanoparticles carrying siRNA could enter Huh7.5.1 cells more efficiently and inhibit the expression of HCV core protein. Conclusion Vitamin E liposome nanoparticles can transport siRNA to Huh7.5.1 cells. And inhibit the expression of HCV core protein.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第二醫(yī)院科研處;重慶醫(yī)科大學(xué)附屬第二醫(yī)院檢驗(yàn)科;重慶醫(yī)科大學(xué)附屬第二醫(yī)院感染科;
【基金】：國(guó)家自然科學(xué)基金面上項(xiàng)目(81171628)~~
【分類(lèi)號(hào)】：R450;R512.63

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