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目的探讨日本血吸虫可溶性虫卵抗原(SEA)对小鼠巨噬细胞株RAW264.7增殖及TNF-α、TGF-β1表达的影响。方法实验分三组:RAW264.7对照组、SEA组和细菌脂多糖(LPS)对照组。培养巨噬细胞株RAW264.7,用20μg/ml SEA分别作用巨噬细胞3、6、9、12和15 h后,荧光定量PCR检测炎症因子TNF-αmRNA和TGF-β1 mRNA的表达;倒置显微镜观察SEA作用12 h后巨噬细胞的形态;Western blot检测炎症相关蛋白TNF-α和TGF-β1的表达;MTT比色法检测SEA对细胞增殖的影响。结果 SEA作用RAW264.7细胞3 h起,TNF-αmR-NA和TGF-β1 mRNA的表达逐渐增高,在第12 h时,其相对表达倍数分别达23.07±3.172和9.83±1.45,与其他各时间点比较差异均有统计学意义(均P<0.05);SEA作用12 h巨噬细胞体积明显增大,细胞拉长且形态不规则;SEA作用巨噬细胞24、48、72 h时,A_(490)值分别与RAW264.7对照组相比,差异均有高度统计学意义(均P<0.01),而与LPS组的差异无统计学意义(P>0.05);在SEA作用RAW264.7细胞12 h时,TNF-α和TGF-β1蛋白表达均高于RAW264.7对照组(均P<0.05)。结论 SEA能促进巨噬细胞的增殖并诱导TNF-α和TGF-ββ1的表达。
Objective To investigate the effects of Schistosoma japonicum soluble egg antigen (SEA) on the proliferation of mouse macrophage cell line RAW264.7 and the expression of TNF-α and TGF-β1. Methods The experiment was divided into three groups: RAW264.7 control group, SEA group and bacterial lipopolysaccharide (LPS) control group. Macrophage cell line RAW264.7 was cultured and macrophages were treated with 20μg / ml SEA for 3h, 6h, 9h, 12h and 15h respectively. The expression of inflammatory cytokines TNF-αmRNA and TGF-β1 mRNA were detected by inverted fluorescence microscope The morphology of macrophages was observed 12 h after SEA treatment. The expression of TNF-α and TGF-β1 were detected by Western blot. The effect of SEA on cell proliferation was detected by MTT assay. Results The expression of TNF-αmR-NA and TGF-β1 mRNA in RAW264.7 cells increased gradually from 3 h after SEA treatment. At 12 h, the relative expression ratios of TNF-αmR-NA and TGF-β1 were 23.07 ± 3.172 and 9.83 ± 1.45, (P <0.05). The volume of macrophages at 12 h after SEA treatment was significantly increased and the cells were elongated and irregular in shape. When macrophages treated with SEA for 24, 48 and 72 h, A_ ( 490) were significantly higher than that of RAW264.7 control group (all P <0.01), but no significant difference with LPS group (P> 0.05). In RAW264.7 cells treated with SEA At 12 h, the expression of TNF-α and TGF-β1 protein were higher than that of RAW264.7 control group (all P <0.05). Conclusion SEA can promote the proliferation of macrophages and induce the expression of TNF-α and TGF-ββ1.