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目的研究蛤蟆草对巨噬细胞多种炎症因子的影响及免疫调节作用。方法通过小鼠血凝抗体滴度检测,考察蛤蟆草乙酸乙酯提取物对体液免疫的作用;通过小鼠迟发型超敏反应试验,考察对细胞免疫的作用;体外试验用ELISA法检测LPS诱导的小鼠巨噬细胞分泌TNF-α、IL-6和NO水平;MTT法检测脾淋巴细胞增殖;Western blot检测磷酸化NF-κB的水平。结果在体试验结果表明蛤蟆草提取物对小鼠体液血凝抗体生成和迟发型超敏反应均具有抑制作用,离体试验也显示其对T淋巴细胞和B淋巴细胞均具有抑制作用。与阴性对照组相比,蛤蟆草组LPS诱导的小鼠巨噬细胞分泌TNF-α、IL-6和NO的水平显著降低,且呈明显的剂量依赖关系。此外,蛤蟆草组磷酸化NF-κB表达水平显著降低。结论蛤蟆草乙酸乙酯提取物对小鼠体液免疫和细胞免疫均具有抑制作用,对TNF-α、IL-6和NO等促炎性细胞因子分泌水平的抑制作用可能是通过抑制NF-κB(p65)的磷酸化所致。
Objective To study the effects of toad on a variety of inflammatory cytokines and immunomodulatory effects in macrophages. Methods The titer of hemagglutination antibody in mice was used to test the effect of the extract of toadstool ethyl acetate on humoral immunity. The effect on the cellular immunity was investigated by the delayed hypersensitivity test in mice. The in vitro test was used to detect the induction of LPS The levels of TNF-α, IL-6 and NO secreted by mouse macrophages were measured. The proliferation of splenic lymphocytes was detected by MTT assay. The level of phosphorylated NF-κB was detected by Western blot. Results The in vivo test results showed that the extracts of Toadstool had inhibitory effects on the formation and delayed-type hypersensitivity of humoral and hemostatic antibodies in mice, and in vitro tests also showed that the extracts of Toadstool had inhibitory effects on T lymphocytes and B lymphocytes. Compared with the negative control group, the levels of TNF-α, IL-6 and NO secreted by LPS-induced murine macrophages were significantly decreased in a dose-dependent manner. In addition, the phosphorylation of NF-κB expression was significantly reduced in toadstool group. CONCLUSION: The extract of toadstool ethyl acetate has inhibitory effect on humoral immunity and cellular immunity in mice. The inhibitory effect on the secretion of TNF-α, IL-6 and NO and other proinflammatory cytokines may be related to inhibition of NF-κB p65) phosphorylation.