siRNA对乙型肝炎孕鼠及胎鼠体内的抗乙型肝炎病毒的保护作用探讨

Discussion of protective effect of anti-hepatitis B virus of siRNA on pregnancy mouse and fetal mouse with infected Hepatitis B virus

目的探讨siRNA干扰对乙型肝炎病毒(HBV)感染孕鼠及胎鼠病毒的清除作用,并了解其阻断垂直传播的可能性。方法体外设计并合成siRNA,扩增,与HBV全基因组质粒pcDNA3.1-DNA注入孕鼠体内为实验组,无意义RNA与pcDNA3.1-DNA注入为对照组。96h后,取孕鼠血液,肝脏及胎血、胎肝,通过血清中HBV-DNA荧光定量,肝组织免疫组化,肝组织HBVC-mRNARTPCR等方法,检测其HBV在体内的表达情况。结果实验组孕鼠血中HBV-DNA定量[(2.77±0.95)Lgcopies/ml]较对照组HBV-DNA定量[(6.89±2.15)Lgcopies/ml]明显减少,差异有统计学意义(P<0.05);实验组孕鼠肝组织HBsAg阳性细胞[(22.5±14.6)个]及HBcAg阳性细胞数[(28.8±17.3)个]均少于对照组的[(42.5±19.2)个、(53.5±23.3)个],差异均有统计学意义(P<0.05);实验组胎鼠肝组织HB-sAg阳性细胞数[(17.5±9.6)个]及HBcAg阳性细胞数[(27.5±15.8)个]均少于对照组[(32.5±15.2)个和(45.7±16.5)个],差异均有统计学意义(P<0.05);孕鼠和胎鼠2组mRNA表达差异均有统计学意义(P<0.05)。结论 siRNA干扰可明显减少孕鼠及胎鼠的肝细胞上HBV表面及核心抗原表达,同时减少了HBVDNA在肝组织中的复制。说明siRNA有减少了母体内HBV病毒,同时减少了垂直传播的可能性。

Objective To investigate the effect on getting rid of hepatitis B virus（HBV） in pregnancy mouse and fetal mouse infected HBV by small interfereing RNA（siRNA） , and understanded the possibility of blocking vertical transmission. Methods siRNA after amplificatd inject into pregnancy mouse, with pcDNA3.1-DNA as experimental group;With nonsense RNA as control group. Blood, liver of pregnancy mouse and of fetal mouse are getted at 96h following injection. Fluorescent quantitation of HBV DNA, liver immunohistochemisty, RT-PCR of HVB mRNA were executed. Results Pregnancy mouse＇s blood HBV-DNA quantitation at experimental group [ （ 2.77 ± 0.95 ） Lg copies/ml ] was less than that at control group [ （6.89 ±2.15 ）Lg copies/ml ] （P 〈 0.05 ）. At pregnancy mouse of experimental group, HBsAg positive cells of liver [ （ 22.5 ± 14.6 ） copy ] and HBcAg positive ceils of liver [ （ 28.8 ± 17.3 ） copy ] were less than those at control group [ （42.5± 19.2 ） copy, （ 53.5 ± 23.3 ） copy ] （ P 〈 0.05 ）. Of fetal mouse, HBsAg positive cells of liver [ （ 17.5 ± 9.6 ） copy ] and HBcAg positive cells of liver [ （27.5 ± 15.8 ）copy ] were less than those at control group [ （32.5± 15.2 ）copy and （45.7 ± 16. 5 ） cop- Y 1 （ P 〈 0. 05 ）. In pregnancy mouse and fetal mouse, mRNA expression of two groups were statistically significant（ P 〈 0.05 ）. Conclusion siRNA can reduce the expression of HBsAg and HBcAg and the replication of HBV DNA in mouse,either pregnancy mouse or fetal mouse. So it may be block up vertical transmission of HBV.