摘要
目的:我们前期研究发现在正常大鼠肝脏中存在2个巨噬细胞亚群,本实验将探讨猪血清诱导免疫性肝损伤大鼠肝脏巨噬细胞的构成及作用。方法:(1)制作大鼠免疫性肝损伤模型。(2)肝脏病理组织学及免疫荧光研究。(3)流式细胞技术研究并分离肝脏巨噬细胞。(4)实时定量PCR检测肝脏巨噬细胞、炎症及纤维化相关细胞因子及Ki67(细胞增殖抗原,MKI67)的表达。结果:(1)猪血清腹腔注射组(IPS组)大鼠肝内大量纤维间隔形成,α平滑肌肌动蛋白(α-SMA)阳性的肌成纤维细胞增生,ED2(CD163,表面抗原分化簇163)阳性细胞增多。(2)免疫荧光及印度墨注射后显示IPS组肝脏中巨噬细胞大小不一,自身荧光强弱不等,在肝内分布不同;GdCl3仅可清除体积大、自身荧光强的细胞。(3)自大鼠肝脏分离出2个巨噬细胞亚群,即ED2染色强阳性、自身荧光强的细胞(ED2high/AFhigh)和ED2染色弱阳性、自身荧光弱的细胞(ED2dim/AFdim),与对照组相比IPS组2个亚群细胞数量均显著增多。(4)ED2high/AFhigh细胞中主要表达与炎症相关的细胞因子,ED2dim/AFdim细胞则主要表达与纤维化相关的细胞因子。(5)ED2dim/AFdim细胞Ki67表达显著高于ED2high/AFhigh细胞,IPS组巨噬细胞Ki67表达显著高于对照组。结论:猪血清诱导免疫性肝脏损伤大鼠肝脏中亦存在2个巨噬细胞亚群,它们在肝脏损伤发病中发挥不同的作用,ED2dim/AFdim细胞增殖活跃,主要表达与纤维化相关的基因,可能在猪血清诱导的肝纤维化形成中起着重要作用。
AIM： Our previous study identified two subsets of liver macrophages in normal rat liver, here we plan to identify and study the liver macrophages in pig serum induced rat liver injury. METHODS ： Immunologically mediated liver injuries were chronically induced in rats by injection of pig sera （IPS）. The histology and immunofluorescent characteristics of the livers were studied. The liver macrophages were isolated and identified by flow cytometry. The mRNA expression of the genes relating to inflammation, fibrogenesis and Ki67 in both subsets of liver macrophages was quantitatively analyzed by the method of quantitative real time PCR （qPCR）. RESULTS： The IPS livers were characterized by intensive septum formation, positive α- SMA ＋ myofibroblast and proliferation of ED2 ^＋ cells. Immunofluorescent image and Indian ink staining showed that the liver macrophages were in different size, different autofluorescence and different distribution. GdCl3 only deleted the liver macrophages of big size/strong autofluorescence. Two subsets of macrophages were isolated and identified from the rat livers, namely ED2^high/AF^high and ED2^high/AF^high, both of which in IPS group increased in number as compared to control group. ED2^high/AF^high subset expressed cytokines mainly related with inflammation, whereas ED2^dim/ AF^dim subset expressed cytokines mainly related with fibrosis. The mRNA level of Ki67 expression was significantly higher in ED2^dim/AF^dim subset than that in ED2^high/AF^high subset, and it was significantly higher in macrophages isolated from IPS group than that from control group. CONCLUSION： The liver macrophages can be divided into two subsets in pig serum induced rat liver injury model. The cells of two subsets play different roles in the pathogenesis of pig serum induced liver injury. The ED2^dim/AF^dim cells proliferate actively and express cytokines mainly related with fibrosis, indicating an imporant role in the formation of pig serum induced liver fibrosis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2010年第7期1341-1347,共7页
Chinese Journal of Pathophysiology
基金
广东省自然科学基金资助项目(No.4009383)
