蝎毒耐热多肽对脂多糖诱导的BV2细胞炎性反应的抑制作用

Inhibitory effect of scorpion venom heat resistant peptide on lipopolysacchrideinduced inflammatory response in BV2 cells

目的观察蝎毒耐热多肽（SVHRP）对细菌脂多糖（LPS）诱导的BV2细胞炎性反应的影响，探讨其是否具有抑制神经炎症的作用。方法：LPS诱导BV2细胞活化，进行免疫细胞化学染色（OX-42抗体）检测细胞的形态，MTT法检测SVHRP对细胞的毒性；分别用Griess试剂法和酶联免疫吸附法（ELISA）测定细胞外液炎性介质一氧化氮（NO）和肿瘤坏死因子-α（TNF-α）的含量；Western Blot检测诱导型NO合成酶（iNOS）蛋白表达水平。结果：不同浓度的SVHRP（2-50μ/ml）对BV2细胞均无毒性。SVHRP（20μg／m1）预处理能明显抑制LPS诱导的BV2细胞的形态活化改变，减少细胞激活后产生的炎性因子NO和TNF-α，抑制iNOS的蛋白表达。结论：SVHRP明显抑制BV2小胶质细胞的炎性反应，提示SVHRP具有抗神经炎症作用。

Objective: To investigate the effect of scorpion venom heat resistant peptide(SVHRP) on lipopolysacchride(LPS)-induced inflammatory response in BV2 cells and decern whether SVHRP has the potential of suppressing CNS inflammation. Methods: LPS was used to induce activation of BV2 cells and changes in BV2 cell morphology were examined by immunocytochemical staining with OX-42 antibody. MTT assay was used to evaluate whether SVHRP was toxic to BV2 cells. Extracellular levels of inflammatory mediators including nitric oxide(NO) and tumor necrosis factor-α(TNF-α) were measured with Griess test and ELISA respectively,and the protein level of inducible NO synthase(iNOS) was examined by Western Blot. Results: SVHRP at doses of 2- 50 μg /ml showed no toxicity to BV2 cells. Pretreatment with20 μg /ml of SVHRP inhibited morphological changes in BV2 cells induced by LPS,and suppressed the production of NO and TNF-α as well as iNOS expression in BV2 cells stimulated with LPS. Conclusion: SVHRP significantly inhibits the inflammatory response in BV2 microglia cells,suggesting that SVHRP may have an anti-inflammatory effect.