摘要
目的通过构建急性乙型肝炎病毒(HBV)小鼠模型,以期探讨La在急性HBV模型中的表达及意义。方法采用水动力法构建急性HBV模型;分别用RT-PCR、ELISA、生化发光检测小鼠血清中HBV DNA、HBsAg、HBeAg和ALT水平;免疫组化和HE染色检测肝中HBcAg和损伤情况;RT-PCR、Western bolt和ELISA检测小鼠La的mRNA水平和蛋白表达水平变化。结果小鼠血清中HBV DNA、HBsAg、HBeAg和肝组织中HBcAg水平显示,急性HBV小鼠模型构建成功;血清ALT水平和HE表明造模后前3 d肝脏损伤较为严重,随后逐渐恢复正常;肝组织中La的mRNA水平提示,HBV模型组普遍高于对照组,而Western bolt结果显示,并无统计学差异,但是血清中La水平普遍高于对照组,尤其是第7天(P<0.000 1)。结论成功构建了急性HBV模型,该模型中急性HBV可增加La蛋白的血清学表达,提示La可能参与了HBV致病过程,或可作为诊断标志物。
OBJECTIVE To explore the role of La protein in acute hepatitis B virus(HBV)by establishing a mouse model.METHODS We constructed an acute hepatitis B virus model by hydrodynamics-based injection of plasmid pAAV/HBV1.2 containing 1.2 times HBV genome.The expression levels of HBV DNA,HBsAg and HBeAg in serum were detected by RT-PCR and ELISA,respectively.HBcAg in liver tissue were assayed by immunohistochemical staining.The level of ALT was detected by bioluminescence.Pathological changes in liver carried by Hematoxylin and eosin(HE)staining.RT-PCR,Western blot and ELISA were used to detect the changes of mRNA level and protein expression level of La protein.RESULTS The levels of HBV DNA,HBsAg and HBeAg in the serum of mice,and the percentage of HBcAg-positive hepatocytes in liver tissues was up to 3.66%at day 5 showed that the mouse model of acute HBV was successfully constructed.The level of serum ALT and HE results showed that the liver damage was serious after the model was established,and then dropped sharplyand returned to normal.The mRNA level of La protein in the liver of the mice indicates that the level of La protein in the model group was generally higher than that in the control group.However,Western bolt results showed that there was no significant difference in intracellular protein levels,but the level of La protein in serum of mice is generally higher thancontrol group,especially on the 7th day(P<0.000 1).CONCLUSION The acute HBV model was successfully constructed.The level of La protein in this model is related to HBV expression,suggesting that La protein may be involved in the pathogenesis of HBVandbea new marker in diagnosis of HBV.
作者
童双梅
潘佳倩
汤静
TONG Shuang-mei;PAN Jia-qian;TANG Jing(Department of Clinical Pharmacy,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China;Department of Pharmacy,The Obstetrics & Gynecology Hospital of Fudan University,Shanghai 200011,China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2019年第12期981-986,共6页
Chinese Pharmaceutical Journal
基金
国家自然科学基金项目资助(81470852)
上海市临床药学重点专科建设项目资助(AB83110002017005)
中国药学会-以岭生物医药创新基金项目资助(GL-B03-20180298)
