内毒素血症大鼠肠黏膜免疫细胞的变化及通腑颗粒的干预作用

摘要

Objective To observe the changes of intestinal mucosal immune cells in rats with endotoxemia and investigate the effects of Tongfu granules on the changes.Methods One hundred and twenty male Wistar rats were randomly divided into 3 groups: control group ( group A), endotoxemia group ( group B) and treatment group ( group C).Group B and C were induced endotoxemia by a single caudal venous injection of 1％ LPS solution (10 mg/kg).Group A was injected normal saline equivalently (1 mL/kg).Group C was given 50％ Tongfu solution (5 g/kg) by a single gastric lavage immediately after LPS solution injected.Group A and B were given distilled water equivalently.Animals were anesthetized and sacrificed in batches (ten animals each time) at 2, 6, 12 and 24 hour after molding.The pathological changes of intestinal mucosa were observed.The numbers of immune cells in the intestinal mucosa were counted after immunohistochemical staining.Results Compared with group A, there were statistically significant decrease of M-cells, DCs, CD8 + T cells, IgA + B cells while increase of CD4 + T cells, Tr cells and apoptotic lymphocytes in group B.There were no statistically significant differences of M-cells, CD8 + T cells and Tr cells between group C and group A.Compared with group A, the count of DCs, IgA + B cells and apoptotic lymphocytes were significantly decreased while CD4 + T cells were increased in group C.There was no statistically significant difference of M-cells between group C and group B.Compared with group B, the count of Tr cells and apoptotic lymphocytes were statistically significant decreased while DCs, CD4 + T cells, CD8 + T cells and lgA + B cells were increased in group C.Conclusion The small intestinal mucosa and immune cells were injured and intestinal lymphocyte apoptosis increased in rats with endotoxemia result in intestinal immunodepression.Tongfu granules can protect the intestinal immune function due to reducing the injury of small intestinal mucosa and immune cells and decreasing intestinal lymphocyte apoptosis.%目的 探讨内毒素血症大鼠肠黏膜免疫细胞的变化及中药通腑颗粒对其影响,为临床治疗提供理论依据.方法 120只雄性Wistar大鼠随机分为正常对照组(A组)、内毒素血症组(B组)及中药治疗组(C组)三组.B组及C组采取尾静脉一次性注射给药方法,注射1%LPS溶液(给药量为10 mg/kg)制作内毒素血症模型,A组注射等量生理盐水(1 mL/kg);C组尾静脉注射给药后即刻给予50%通腑颗粒溶液(给药量为5 g/kg)灌胃,A组及B组给予等量蒸馏水灌胃.造模后在第2、6、12、24 h时间点分批(每次10只)麻醉后处死动物,取小肠组织进行检测.采用HE染色观察肠黏膜病理变化,采用免疫组织化学及TUNEL染色方法,分别标记并计数各组大鼠肠黏膜免疫细胞及凋亡淋巴细胞.结果 与A组比较,B组肠黏膜M细胞、DC细胞、CD8+T细胞及IgA+B细胞计数明显减少,而CD4+T细胞、Tr细胞及淋巴细胞凋亡计数明显增多;C组肠黏膜M细胞、CD8+T及Tr细胞计数无明显改变,DC细胞、IgA+B细胞及淋巴细胞凋亡计数明显减少,而CD4+T细胞计数明显增多;与B组比较,C组肠黏膜M细胞无明显改变,Tr细胞及淋巴细胞凋亡计数明显减少,而DC细胞、CD4+T细胞、CD8+T细胞及IgA+B细胞计数明显增多.结论 内毒素血症时肠黏膜损伤,肠免疫不同阶段的免疫细胞均发生损害,Th1/Th2比例失调,淋巴细胞凋亡增多,导致肠黏膜局部免疫发生抑制.应用通腑颗粒可以减轻肠黏膜损伤,减轻免疫细胞损伤,改善Th1/Th2比例,减少淋巴细胞凋亡,保护局部免疫功能.