高表达线粒体融合蛋白2(MFN2)可抑制高BHBA活化的奶牛肝细胞NF-κB炎性通路

Overexpression of MFN2 inhibits the activation of NF-κB inflammatory pathway induced by BHBA in cow hepatocytes

为研究线粒体融合蛋白2(mitofusin 2,MFN2)对高β-羟丁酸(β-hydroxybutyrate,BHBA)活化NF-κB炎性通路的影响,体外培养奶牛肝细胞,添加不同浓度(0.0,1.2,2.4,4.8 mmol/L)的BHBA,并转染过表达MFN2的腺病毒,运用Western blot和qRT-PCR技术检测NF-κB炎性通路关键分子的基因和蛋白表达。结果显示,随着BHBA浓度的增加,IKBα和NF-κB p65的磷酸化水平以及IL-1β、IL-6和TNFα的mRNA表达均显著升高,MFN2的基因和蛋白表达水平则显著降低;过表达MFN2后,显著抑制了高BHBA诱导的IKBα和NF-κB p65磷酸化水平及IL-1β、IL-6、TNFαmRNA表达水平的升高。结果表明,在奶牛肝细胞中过表达MFN2可以显著抑制高BHBA活化的NF-κB炎性通路。

This aim of this study was to investigate the effect of mitofusin 2(MFN2)on the activation of NF-κB signaling pathway induced by high concentration ofβ-hydroxybutyrate(BHBA)in cow hepatocytes.Hepatocytes were isolated from healthy calves and were treated with different concentrations of BHBA(0.0,1.2,2.4,4.8 mmol/L),or adenovirus-mediated overexpression of MFN2.Western blot and qRT-PCR were used to detect the changes of mRNA and protein expression of molecules involved in NF-κB pathway.The results showed that the phosphorylation of IKBαand NF-κB p65,and the mRNA expression of IL-1β,IL-6,and TNFαwere significantly increased,while the expression of MFN2 was significantly decreased with the increased concentration of BHBA.However,overexpression of MFN2 significantly inhibited the phosphorylation of IKBα,NF-κB p65 and the expression of IL-1β,IL-6 and TNFαmRNA induced by BHBA.In summary,overexpression of MFN2 in cow hepatocytes could inhibit the activation of NF-κB inflammatory pathway induced by high concentration of BHBA.