[关键词]
[摘要]
目的 研究莫诺苷对肿瘤坏死因子-α(TNF-α)诱导成骨细胞MC3T3-E1炎症的保护作用,探讨Caspase、MAPK和NF-κB通路调控机制。方法 TNF-α(50 ng/mL)诱导MC3T3-E1细胞,制备细胞炎症模型;MTT法检测莫诺苷对MC3T3-E1细胞增殖的影响,并检测不同浓度的莫诺苷对TNF-α诱导MC3T3-E1细胞活力的影响;ELISA法检测细胞上清液中炎性因子白细胞介素-1β(IL-1β)、IL-6的水平;Western blotting法检测Caspase-3、Caspase-9、p-ERK、p-JNK、p-p38、p-IκBα、IκBα和NF-κB等相关蛋白的表达。结果 莫诺苷能够促进正常MC3T3-E1细胞增殖,TNF-α诱导MC3T3-E1细胞增殖受到抑制,而莫诺苷对TNF-α诱导MC3T3-E1细胞具有保护作用;莫诺苷可以抑制TNF-α诱导MC3T3-E1细胞IL-1β和IL-6的释放;降低Caspase-3、Caspase-9、p-ERK、p-JNK、p-p38、p-IκBα蛋白的表达,升高IκBα蛋白表达,对NF-κB p65无明显影响。结论 莫诺苷对TNF-α诱导MC3T3-E1炎症细胞有保护作用,其机制为抑制炎性因子的释放、抑制MAPK和NF-κB通路的激活、抑制凋亡蛋白Caspase的表达,从而抑制MC3T3-E1细胞的凋亡。
[Key word]
[Abstract]
Objective To investigate the effects of morroniside extracted from root-bark of Sambucus williamsii on TNF-α-induced MC3T3-E1 cell inflammation, and explore its mechanism of action including Caspase, MAPK, and NF-κB signaling pathway. Methods MC3T3-E1 cells were cultured in medium containing TNF-α (50 ng/mL) and different doses of morroniside. The cell viability was examined by MTT assay, and the levels of IL-1β and IL-6 in the supernatants were identified by ELISA. The expressions of Caspase-3, Caspase-9, p-ERK, p-JNK, p-p38, p-IκBα, IκBα, and NF-κB on the protein level was tested by Western blotting. Results MTT assay results showed that morroniside could promote the proliferation of MC3T3-E1 cells and protect the MC3T3-E1 cells induced by TNF-α. ELISA results showed that the expressions of IL-1β and IL-6 were inhibited. Meanwhile, morroniside could reduce the expression of Caspase3, Caspase9, p-ERK, p-JNK, p-p38, and p-IκBα protein, and increase IκBα protein expression while there was no significant difference in the expression of NF-κB p65. Conclusion Morroniside has protective effect on TNF-α-induced MC3T3-E1 cells inflammation. The possible related mechanism is that morroniside inhibits the release of inflammatory cytokines, the activation of MAPK and NF-κB pathway, and the expression of Caspase, thereby inhibiting the apoptosis of MC3T3-E1 cells.
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[基金项目]
国家自然科学基金面上项目(81173501);哈尔滨市应用技术研究与开发项目计划(2014AB3AS045)