短链脂肪酸通过抑制白细胞介素17A和NF-κB信号通路减轻γδT细胞介导的炎症反应

Short-chain fatty acids alleviate γδT cell-mediated inflammatory response via inhibiting IL-17A and NF-κB signaling pathway

目的 探究短链脂肪酸减轻γδT细胞介导炎症反应的作用机制。方法 使用一定浓度的短链脂肪酸干预大鼠肠道来源的γδT细胞,CCK-8检测短链脂肪酸对γδT活性的影响,酶联免疫吸附测定(ELISA)检测白细胞介素17A(IL-17A)因子含量,实时荧光定量(RT-q PCR)检测IL-17A因子的表达水平,流式细胞仪分析IL-17+γδT细胞的比例,Western blot研究γδT细胞IL-17A和核转录因子κB(NF-κB)信号通路的影响。结果 CCK-8结果提示乙酸钠的浓度≤0.5 mmol/L,丙酸钠、丁酸钠和混合短链脂肪酸的浓度≤0.25 mmol/L对γδT细胞的增殖无抑制作用(P> 0.05);ELISA和RT-q PCR结果显示,丙酸钠、丁酸钠及混合短链脂肪酸处理的γδT细胞IL-17A的含量较Control组均显著降低。流式细胞检测结果示IL-17+γδT细胞的比例在丙酸钠、丁酸钠、混合短链脂肪酸及TSA干预下均明显下降(P <0.001);Western blot检测发现丙酸钠、丁酸钠和混合短链脂肪酸可抑制IL-17A、IKK的表达及NF-κB磷酸化水平(P <0.05)。结论 短链脂肪酸能抑制γδT细胞IL-17A和NF-κB信号通路的激活,从而减轻机体的炎症反应。

Objective To explore the mechanism of short-chain fatty acids in alleviating γδT cell-mediated inflammatory response.Methods A certain concentration of short-chain fatty acids was used to intervene in rat intestinal-derived γδT cells.CCK-8 was used to detect the effect of shortchain fatty acids on γδT activity.ELISA was used to detect the content of the IL-17A,RT-qPCR was utilized to detect the expression of the IL-17A,and flow cytometry was applied to analyze the IL-17+γδT cell frequency.The effect of SCFAs on IL-17A and NF-κB signaling pathway in γδT cells were observed by Western blot.Results CCK-8 assay showed that the concentration of sodium acetate was less than or equal to 0.5 mmol/L,and the concentrations of sodium propionate,sodium butyrate,and mixed short-chain fatty acids were less than or equal to 0.25 mmol/L,indicating no inhibitory effect on the proliferation of γδT cells.ELISA and RT-qPCR showed that the treatment of γδT cells with sodium propionate,sodium butyrate,and mixed short-chain fatty acids significantly decreased the levels of IL-17A compared with the control group.Flow cytometry revealed a significant decrease in the proportion of IL-17+γδT cells under the treatment of sodium propionate,sodium butyrate,mixed shortchain fatty acids,and TSA intervention.Western blot found that sodium propionate,sodium butyrate,and mixed shortchain fatty acids could inhibit the expression of IL-17A and IKK,as well as the phosphorylation level of NF-κB.Conclusion Shortchain fatty acids could inhibit the activation of IL-17A and NF-κB signaling pathway in γδT cells,thereby alleviating the inflammatory response in the body.