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目的:探索枯否细胞在树鼩感染乙肝病毒(HBV)慢性化过程中的意义。方法:树鼩分为3组:A组6只,为前期实验已确定慢性感染HBV的树鼩;B组3只,为疑似慢性感染HBV的树鼩;C组4只,为未接种HBV的正常对照树鼩。全部动物定期抽血和进行肝活检手术;对手术切取的树鼩肝组织进行枯否细胞的分离、纯化和原代培养,采用流式细胞术、细胞免疫组化、溶酶体荧光探针及实时荧光定量RT-PCR等方法检测CD163+细胞数量、溶酶体数量、溶菌酶的表达及肿瘤坏死因子α(TNF-α)mRNA表达水平。结果:(1)慢性感染HBV的树鼩肝脏枯否细胞比例及肝组织内CD163+细胞数量显著高于其它2组(均P<0.05);(2)慢性感染HBV的树鼩肝脏枯否细胞的溶酶体荧光强度、肝组织内溶菌酶阳性细胞计数和TNF-αmRNA的表达水平均显著低于其它2组(均P<0.05)。结论:枯否细胞在宿主感染HBV的慢性化过程中可能起一定的调节作用。
Objective: To explore the significance of Kupffer cells in the process of chronic hepatitis B virus infection. Methods: The tree shrews were divided into 3 groups: 6 in group A, which had been identified as chlamydia of chronically infected HBV in the previous experiment; 3 in group B as tree shrews suspected of chronic HBV infection; 4 in group C, Normal control tree 鼩. All animals were routinely bled and liver biopsy was performed. The liver tissues of tree shrews were excised and purified from Kupffer cells, and primary culture was performed. Flow cytometry, immunohistochemistry, lysosomal fluorescent probes and The number of CD163 + cells, the number of lysosomes, the expression of lysozyme and the mRNA expression of tumor necrosis factor-α (TNF-α) were detected by real-time fluorescence quantitative RT-PCR. Results: (1) The proportion of liver Kupffer cells and the number of CD163 + cells in the liver tissues of chickens infected with HBV were significantly higher than those in the other two groups (all P <0.05). (2) The percentage of liver Kupffer cells Lysosome fluorescence intensity, Lysozyme positive cells count and TNF-αmRNA expression level in liver tissue were significantly lower than the other two groups (all P <0.05). Conclusion: Kupffer cells may play a regulatory role in the process of chronic HBV infection in host.