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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(2~50μg/ml)对BV2细胞均无毒性。SVHRP(20μg/ml)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 observe the effects of scorpion venom heat-resistant polypeptide (SVHRP) on inflammatory reaction induced by bacterial lipopolysaccharide (LPS) in BV2 cells and explore whether it has the effect of inhibiting neuroinflammation. Methods: The activation of BV2 cells was induced by LPS and the morphology of cells was detected by immunocytochemistry (OX-42 antibody). The cytotoxicity of SVHRP was detected by MTT assay. The levels of extracellular load were determined by Griess reagent and enzyme-linked immunosorbent assay (ELISA) The levels of nitric oxide (NO) and tumor necrosis factor-α (TNF-α) were detected by Western Blot. The expression of inducible NO synthase (iNOS) protein was detected by Western Blot. Results: Different concentrations of SVHRP (2 ~ 50μg / ml) were non-toxic to BV2 cells. Pretreatment with SVHRP (20μg / ml) could significantly inhibit the morphological activation of BV2 cells induced by LPS, reduce the production of inflammatory cytokines NO and TNF-α and inhibit the expression of iNOS. Conclusion: SVHRP can significantly inhibit the inflammatory response of BV2 microglia, suggesting that SVHRP has neuroinflammation.