论文部分内容阅读
目的探索慢性乙型肝炎病毒(HBV)感染树鼩Kupffer细胞Toll样受体(TLR)家族中的TLR2和TLR4在mRNA水平的表达情况及其对Kupffer细胞功能的影响。方法树鼩分为确定慢性感染HBV的树鼩、疑似慢性感染HBV的树鼩和未接种HBV的正常对照树鼩。全部动物定期抽血和进行肝活检手术,采用实时荧光定量PCR(qRT-PCR)分析血清和肝组织的HBV DNA水平;对手术切取的树鼩肝组织进行Kupffer细胞的分离、纯化和原代培养,采用qRT-PCR检测TLR2、TLR4以及TNF-α的mRNA表达水平;采用迁移实验及溶酶体荧光探针等方法分析TLR2和TLR4对Kupffer细胞迁移能力及溶酶体数量的影响。结果确定慢性感染HBV的树鼩TLR2 mRNA和TLR4 mRNA表达水平均低于疑似慢性感染HBV的树鼩和未接种HBV的正常对照树鼩(P<0.05),表达水平均与动物肝组织的HBV DNA拷贝数呈负相关(P<0.05),与Kupffer细胞的细胞迁移数、溶酶体密度及TNF-αmRNA表达水平呈正相关(P<0.05)。结论 Kupffer细胞中的TLR2和TLR4可能通过影响Kupffer细胞功能而参与树鼩HBV感染后肝脏病变的慢性化发展过程。
Objective To investigate the expression of TLR2 and TLR4 in the Toll-like receptor (TLR) family of Chlamydia trachomatis Kupffer cells infected by chronic hepatitis B virus (HBV) and its effect on the function of Kupffer cells. Methods Tree shrews were divided into tree shrews to identify chronic HBV infection, tree shrews that were suspected of chronic HBV infection, and tree shrews that were not inoculated with HBV. All animals were routinely drawn for blood and liver biopsy. HBV DNA levels in serum and liver tissue were analyzed by real-time quantitative PCR (qRT-PCR). Kupffer cells were isolated, purified and primary cultured . The mRNA expression of TLR2, TLR4 and TNF-α were detected by qRT-PCR. The migration ability and lysosome number of TLR2 and TLR4 were analyzed by migration assay and lysosome fluorescence probe. The results showed that the expression levels of TLR2 mRNA and TLR4 mRNA in the tree shrews infected with chronic HBV were lower than those in the chickens suspected of chronic HBV infection and in the normal control tree without HBV vaccination (P <0.05) (P <0.05), which was positively correlated with cell migration number, lysosome density and TNF-αmRNA expression in Kupffer cells (P <0.05). Conclusions TLR2 and TLR4 in Kupffer cells may participate in the chronic development of liver pathology after the infection of tree 鼩 with HBV by influencing the function of Kupffer cells.