HBV转基因小鼠尿液中乙肝病毒相关指标的分析  

Analysis of hepatitis B virus-related indexes in the urine of HBV transgenic mice

作　　者：杨阳[1,2] 刘光泽[2] 李秀梅[2] 孔祥平[2] 顾为望[1] 

机构地区：[1]南方医科大学比较医学研究所暨实验动物中心,广州510515 [2]解放军第458医院全军肝病中心,广州510602

出　　处：《中国比较医学杂志》2016年第6期53-60,共8页Chinese Journal of Comparative Medicine

基　　金：国家科技重大专项课题-艾滋病和病毒性肝炎等重大传染病防治-HBV细胞和动物模型的建立和应用(课题编号:2012ZX10004503010)

摘　　要：目的检测分析HBV转基因小鼠尿液中HBV相关指标表达情况,进一步认识转基因小鼠的生物学特性;并通过多种实验方法确定尿液中HBV相关指标的组织来源。方法使用酶联免疫吸附法（ELISA）和荧光定量PCR（real-time PCR）检测转基因小鼠尿液中HBV相关指标;并通过水动力转染的小鼠实验、RNA干扰方法抑制HBV表达转基因鼠实验、乙肝患者HBV阳性血清感染正常小鼠实验等多种实验方法确定其复制表达组织来源。结果 HBV转基因小鼠尿液中存在HBsAg、HBe Ag、及HBV-DNA表达,其中HBsAg的表达水平较高（6674±619.8 IU/m L）,但低于血清中HBsAg的表达水平（16470±2704 IU/m L）,尿液HBsAg雄性小鼠的表达水平高于雌性小鼠;而尿液HBe Ag表达水平较低,且尿液中HBe Ag的阳性率高于血液,HBe Ag的表达水平个体间和性别间有明显差异;尿液中存在HBV-DNA含量达到10~3~10~5copy/m L;未检测到相关抗体表达。通过水动力转染等实验表明转基因鼠尿液中的HBV相关指标来源于肾脏的复制表达,而不是由肝脏分泌进入血液循环以尿液排出,肾脏是HBV独立的表达场所。结论转基因小鼠尿液中存在HBV相关指标的表达,并随月龄长期表达,其中HBsAg及HBV-DNA表达水平较高且比较稳定,雄性小鼠尿液HBsAg的滴度高于雌性小鼠;HBe Ag雄性鼠表达水平相对雌性鼠较高且比较稳定;尿液中无各类抗体表达;肾脏也是HBV独立的复制表达场所。Objective To detect and analyze the HBV-related indexes in the urine of HBV transgenic mice and further understand the biological characteristics of transgenic mice,and to clarify the tissue sources of HBV-related indexes. Methods HBV-related indexes in the urine of transgenic mice were tested using enzyme-linked immune sorbent assay（ ELISA） and fluorescence quantitative PCR（ real-time RCR）. The tissue sources were confirmed by several experiments,i. e. hydrodynamic transfection of mice,RNA interference to inhibit HBV-expression in the transgenic mice,and to infect normal mice with HBV-positive serum from patients. Results Expression of HBsAg,HBe Ag and HBV-DNA was present in the urine of transgenic mice,of which the HBsAg expression level was high（ 6674 ± 619. 8 IU / m L）,butlower than that in the serum（ 16470 ± 2704 IU / m L）. The level of HBsAg expression in the urine of male mice was higher than that in female mice. The level of HBe Ag expression in the urine was lower and the HBe Ag positive rate of urine was higher than that of blood, and the levels of HBe Ag expression showed significant inter-individual and inter-sexual differences. HBV-DNA level reached 103- 105 copy / m L in the urine,but no related antibody expression was detected. The experiments such as hydrodynamic infection test indicated that the HBV-related indexes in the urine are derived from replication in the kidneys rather than secreted from the liver,entered into the blood circulation,and discharged from the urine. The kidneys are an independent expression site of HBV. Conclusions The expression of HBV-related indexes is present in the urine of transgenic mice and it is a long-term expression along with the age in months,of which the expression levels of HBsAg and HBV-DNA are rather high and stable. HBsAg titer in the urine of the male mice is higher than that of female mice. HBe Ag expression level in the male mice is more stable compared with that in female mice. No expressions of various kinds of antibodies have been fo

关 键 词：肝炎病毒 乙型 转基因小鼠 模型 动物 尿液 生物学特性 

分 类 号：R33[医药卫生—人体生理学]