日本血吸虫可溶性虫卵抗原对体外培养大鼠腹腔巨噬细胞活化及TNF-α表达的影响  

Effect of soluble egg antigen of Schistosoma japonicum on activation and expression of TNF-α in cultred rat peritoneal macrophages in vitro

作　　者：胡敏[1] 王亚东[1] 黄顺[1] 钱琛[1] 

机构地区：[1]安徽中医药大学病理学教研室,安徽合肥230038

出　　处：《皖南医学院学报》2015年第3期209-213,共5页Journal of Wannan Medical College

基　　金：安徽省高等学校省级优秀青年人才基金(2009SQRZ118)

摘　　要：目的:观察日本血吸虫可溶性虫卵抗原(SEA)对体外培养的大鼠腹腔巨噬细胞(PMCs)中TNF-α活化及表达的影响。方法:体外培养大鼠PMCs,利用苏木素-伊红染色(HE染色)、免疫细胞化学染色(IHC)、RT-PCR以及放免法,对SEA作用后的大鼠PMCs形态变化和TNF-α表达进行观察;同时,制备SEA活化腹腔巨噬细胞条件培养基(SEA-MCM),利用MTT比色法和3H-脯氨酸掺入法,对经过SEA-MCM作用后的HSC-T6细胞增殖及胶原合成结果进行检测。结果:1大鼠PMCs在LPS(10 mg/L)和SEA(10 mg/L)分别作用3 h、6 h和9 h后,可以使PMCs活化增加,并促进大鼠PMCs TNF-α蛋白表达(P<0.05);2LPS(10 mg/L)和SEA(10 mg/L)分别作用6 h,大鼠PMCs TNF-αmRNA表达增加(P<0.05);3LPS(10 mg/L)和SEA(10 mg/L)分别作用3 h、6 h和9 h,培养上清中TNF-α含量高于阴性对照组(P<0.05);6 h和9 h组均高于3 h组(P<0.05),6 h和9 h组间无明显差别(P>0.05);4SEA-MCM促进HSC-T6细胞增殖和胶原合成(P<0.05)。结论:体外实验证实SEA与LPS在活化大鼠PMCs、促进TNF-α表达和分泌中有相似的作用,这可能与SEA活化大鼠PMCs能明显促进HSC-T6细胞增殖和胶原合成有关。Objective： To observe the effect of soluble egg antigen（ SEA） on TNF-α expression in rat peritoneal macrophages in vitro and proliferation and collagen synthesis of HSC-T6 cells. Methods： Rat peritoneal macrophages were harvested via irrigation of the peritoneal cavity,and cultured in vitro. The effect of SEA on the expression of TNF-α in the rat peritoneal macrophages was observed after staining of the cells with HE and immunocytochemistry,RTPCR and radiommunity. SEA-MCM was prepared,and MTT colorimetric assay and 3H-proline incorporation were performed to detect the proliferation and collagen synthesis of HSC-T6. Results： ①At 3,6 and 9h,the expression of TNF-α in the SEA and LPS group was significantly higher than that in the control treated with RPMI1640（ P〈0. 05）; ②The level of TNF-α in the supernatant was also significantly higher than that in the control treated RPMI1640（ P〈0. 05）; ③After exposure to SEA or LPS for 3,6 and 9 h,the expression of TNF-α mRNA of peritoneal macrophages in the SEA and LPS group was significantly higher than that in the control after treatment with RPMI1640（ P〈0. 05）; ④SEA-MCM significantly promoted the proliferation and collagen production of HSC-T6 cells（ P〈0. 05）. Conclusion： SEA,functioning similarly with LPS,may have the effect to induce rat peritoneal macrophages activation and promote expression and secretion of TNF-α,which is possibly related to the capacity of SEA that is capable of promoting the proliferation and collagen production of HSC-T6 in vitro.

关 键 词：腹腔巨噬细胞 肿瘤坏死因子-α 血吸虫可溶性虫卵抗原 增殖 胶原合成 

分 类 号：R392[医药卫生—免疫学] R575.2[医药卫生—基础医学]