花生四烯酸氨基乙醇对人成牙本质细胞样细胞MMP-9表达的影响

Regulation of matrix metalloproteinase-9 expression by anandamide in human odontoblast-like cells

目的探讨花生四烯酸氨基乙醇(anandamide,AEA)对人成牙本质细胞(HODs)样细胞中基质金属蛋白酶(MMP)-9表达的影响。方法体外培养HODs样细胞,免疫荧光进行形态和功能验证,四甲基偶氮唑盐(MTT)法研究AEA对HODs样细胞活性影响,实时荧光定量聚合酶链反应(RT-qPCR)和Western blotting检测AEA处理后MMP-9基因和蛋白表达水平及AEA的作用受体大麻素受体1,2和TRPV1的表达,并检测MAPK信号通路的调控作用。结果大麻素受体1,2和TRPV1在HODs样细胞中表达,不同浓度AEA和不同处理时间可刺激MMP-9基因和蛋白表达水平提高。抑制大麻素受体1,2和TRPV1后AEA诱导的MMP-9表达水平下降,其中大麻素受体1和TRPV1作用较强。JNK信号通路在AEA对MMP-9的表达调节中起主导作用。结论AEA主要通过大麻素受体1和TRPV1调控HODs样细胞中MMP-9的表达,JNK信号通路是此过程的主要调节通路。

Objective To investigate the anandamide-induced expression of matrix metalloproteinase(MMP)-9 in human odontoblasts(HODs)-like cells.Methods HODs-like cells were cultured in vitro,and their morphology and function were verified by immunofluorescence.The effect of anandamide on cell viability of HODs-like cells was examined by MTT assay.The gene and protein expression levels of MMP-9,cannabinoid receptors 1,cannabinoid receptors 2 and transient receptor potential vanilloid-1(TRPV1)after different treatments were measured by real-time quantitative fluorescence polymerase chain reaction(RT-qPCR)and Western blotting.Results Cannabinoid receptors 1,cannabinoid receptors 2 and TRPV1 were expressed in HODs-like cells.Increased MMP-9 expression at gene and protein levels were found when treated with different concentrations or time of anandamide.After inhibition of cannabinoid receptors 1,cannabinoid receptors 2 and TRPV1,the anandamide-induced MMP-9 expression decreased,especially when inhibiting cannabinoid receptors 1 and TRPV1.JNK signaling pathway played a predominant role in the regulation of anandamide on MMP-9.Conclusions The upregulation effect of anandamide on MMP-9 expression in HODs-like cells was mainly through the cannabinoid receptors 1 and TRPV1.This process was regulated mainly by the JNK signaling pathway.