树鼩慢性感染乙肝病毒过程中枯否细胞变化的意义*

 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-α mRNA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。

Changes of Kupffer cells during tree shrew chronically infected with hepatitis B virus

AIM：To explore the changes and significance of Kupffer cells in the process of tree shrew chronically infected with hepatitis B virus (HBV). METHODS：The animals were divided into 3 groups. Group A consists of 6 tree shrews that were identified as persistently infected with HBV; group B consists of 3 tree shrews that were suspected as persistently infected with HBV; group C consists of 4 tree shrews that were not inoculated with HBV and were applied as normal controls. Liver biopsies were collected regularly from all animals, and the Kupffer cells were isolated, purified and primarily cultured. The techniques of flow cytometry, immunohistochemistry, lysosomal fluorescent probe staining and real-time RT-PCR were applied to determine the number and function of these Kupffer cells. RESULTS：The result showed that the count and proportion of CD163+ cells in group A were significantly higher than those in group B and group C (P<0.05). Meanwhile, the fluorescence intensity levels of lysosomal, the number of lysozyme-positive cells and the mRNA expression level of TNF-α in the Kupffer cells in group A were significantly lower than those in group B and group C (P<0.05). CONCLUSION：Kupffer cells may play a regulatory role during host’s chronic HBV infection.