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目的观察TLR4和Notch1信号交叉调控炎症以及芝麻素的干预作用。方法体外培养RAW264.7细胞,MTT法检测LPS和芝麻素对RAW264.7细胞毒性作用,Reverse transcription-PCR(RT-PCR)观察LPS刺激后RAW264.7细胞Notch信号相关基因Jagged1,Notch1和Hes1的表达,ELISA和Western blot方法检测芝麻素和Notch1抑制剂DAPT对RAW264.7细胞合成和释放白细胞介素-6(IL-6)和肿瘤坏死因子α(TNF-α)的影响。结果 200 ng/ml LPS处理2-6 h后能明显提高Jagged1,Notch1和Hes1基因的表达(P<0.05),其中4h最为明显(P<0.01)。同时,LPS明显刺激RAW264.7合成和释放IL-6,TNF-α(P<0.01),芝麻素和Notch1抑制剂GAPT能明细降低LPS诱导RAW264.7细胞IL-6,TNF-α的分泌(P<0.05)。结论 TLR4和Notch1信号交叉调控炎症细胞因子产生,芝麻素能够抑制TLR4和Notch1信号介导的炎症反应。
Objective To observe the effects of interleukin-4 (TLR4) and Notch1 signaling on inflammation and sesamin intervention. Methods RAW264.7 cells were cultured in vitro. MTT assay was used to detect the cytotoxicity of LPS and sesamin on RAW264.7 cells. Reverse transcription-PCR (RT-PCR) was used to detect the expression of Notch signal related genes Jagged1, Notch1 and Hes1 in RAW264.7 cells The effects of sesamin and Notch1 inhibitor DAPT on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in RAW264.7 cells were detected by ELISA and Western blot. Results The expression of Jagged1, Notch1 and Hes1 genes was significantly increased after treated with 200 ng / ml LPS for 2 to 6 hours (P <0.05), and the most obvious was in 4 hours (P <0.01). At the same time, LPS significantly stimulated the synthesis and release of IL-6 and TNF-α by RAW264.7 (P <0.01). Sepsis and Notch1 inhibitor GAPT attenuated the secretion of IL-6 and TNF-α induced by LPS in RAW264.7 cells P <0.05). Conclusions TLR4 and Notch1 signals cross-regulate the production of inflammatory cytokines. Sesamin can inhibit the inflammatory response mediated by TLR4 and Notch1 signals.