灵芝多糖对H22肝癌小鼠肠道黏膜免疫功能的影响

作者	单位
周桂琴	北京地坛医院中西医结合科
赵宏艳	中国中医科学院中医基础理论研究所
吕诚	中国中医科学院中医临床基础医学研究所
张皖东	安徽中医学院第一附属医院
谭勇	首都医科大学中医药学院
陈士林	广东省深圳市中药药学及分子药理学研究重点实验室
杨大坚	广东省深圳市中药药学及分子药理学研究重点实验室
陈新滋	广东省深圳市中药药学及分子药理学研究重点实验室
吕爱平	中国中医科学院中医临床基础医学研究所

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中文摘要:

目的观察灵芝多糖(ganoderma lucidum polysaccharides,GLP)抗肿瘤的黏膜免疫机制。方法将H22细胞浓度调整为1×109个/L,于昆明小鼠右腋下行无菌皮下接种,每只小鼠接种0.2mL细胞悬液,接种后随机将动物分为模型组、GLP组、环磷酰胺(cytoxan,CTX)组、GLP+CTX组,另设空白对照组,每组8只。于造模后第2天,GLP组及GLP+CTX组分别以1.02g/(kg.d)灌胃GLP,每天1次,连续12天;CTX组及GLP+CTX组分别于第1天及第6天按100mg/(kg.d)腹腔注射CTX;空白对照组与模型组以等量蒸馏水灌胃。于造模后第14天,拉颈处死小鼠,分离小肠,取盲肠上端1cm处回肠,4%多聚甲醛固定,石蜡切片,采用免疫组化(iHC)的方法检测免疫球蛋白A(immunoglobulin A,IgA)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-2(interleukin-2,IL-2)及IL-10的表达;剩余小肠分离肠道上皮内淋巴细胞(intraepithelial lymphocytes,IEL)、固有层内淋巴细胞(lamina propria lymphocytes,LPL)及PP结内淋巴细胞(peyer′s patch lymphocytes,PPL),采用免疫荧光染色及流式细胞仪分析T淋巴细胞亚群的变化。结果(1)与正常组比较,模型组肠道黏膜淋巴细胞表型及肠道内细胞因子的表达均发生改变,并且各部位的淋巴细胞表型变化不同;(2)与模型组比较,GLP、CTX及GLP+CTX均能不同程度的调节肠道黏膜淋巴细胞的表型及细胞因子的表达。结论GLP对肠道黏膜免疫的调节作用可能是其发挥抑瘤作用的另一条途径。

英文摘要:

Objective To observe the mucosal immune mechanism of anti-tumor action of Ganoderma lucidum polysaccharides(GLP).Methods The concentration of H22 cells in suspension were adjusted to 1×109/L,and 0.2 mL of the cell suspension was injected subcutaneously in the right oxter of Kunming mice.Then the H22 bearing mice were randomly divided into 4 groups:the GLP group,the Cytoxan(CTX) group,the CTX+GLP group and the untreated model group,8 mice in each group.Besides,a blank control group was set up.Starting from the 2nd day of modeling,GLP,at the dose of 1.02 g/kg was given to GLP group and GLP+CTX group by gastrogavage once a day for 12 successive days;CTX at the dose of 100 mg/kg was administered via peritoneal injection to the CTX group and the GLP+CTX group on the 1st day and the 6th day of the experimental course;but to the model group and the blank group,only equal volume of distilled water was given.All mice were sacrificed on the 14th day,the ileum at 1 cm upper to cecum was taken,through 4% paraform fixation and paraffin section,it was used for immunohistochemical detecting expressions of immunoglobulin A(IgA),tumor necrosis factor-α(TNF-α),interleukin-2(IL-2) and interleukin-10(IL-10) in ileum.Besides,the lymphocyte subsets in the intraepithelial lymphocyte(IEL),lamina propria lymphocytes(LPL),and Peyer’s patch lymphocytes(PPL) were analyzed by immune fluorescence technique and flow cytometry.Results Compared with the blank control group,the phenotype of lymphocytes and the expression of cytokines in ileum in the model group changed significantly;and the phenotype was variant in different regions.Compared with the model group,both indexes were adjusted in the GLP,CTX and GLP+CTX group to different degrees.Conclusion The adjustment of GLP on intestinal mucosal immune is probably another path for its anti-tumor action.

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