摘要
目的研究和厚朴酚(HNK)对类风湿关节炎滑膜细胞(FLS)活化的抑制作用及其分子机制。方法将肿瘤坏死因子(TNF)-α处理MH7A细胞建立FLS活化模型,细胞外调节激酶(ERK)抑制剂MK-8353(MK)抑制ERK激活,siRNA干扰抑制肿瘤坏死因子受体相关因子3(TRAF3)表达。采用CCK-8检测细胞增殖,ELISA检测白细胞介素(IL)-6和CXC趋化因子配体10(CXCL10)分泌情况,Western blot检测基质金属蛋白酶(MMP)-2、MMP-9、TRAF3和p-ERK蛋白表达,Real-time PCR检测TRAF3 mRNA表达。结果TNF+HNK组细胞增殖较CON组和TNF组明显降低,且IL-6、CXCL10分泌和MMP-2、MMP-9蛋白表达较TNF组明显降低(P<0.05)。与CON组和TNF组相比,TNF+HNK组TRAF3 mRNA和蛋白表达均明显升高(P<0.05)。si-TRAF3组TRAF3蛋白表达较si-CON组明显降低(P<0.05),TNF+HNK+si-TRAF3组IL-6、CXCL10分泌和MMP-2、MMP-9蛋白表达较TNF+HNK组明显升高(P<0.05)。与CON组和TNF组比较,TNF+HNK组p-ERK1/2蛋白表达明显升高(P<0.05),TNF+HNK+MK组p-ERK1/2、TRAF3蛋白表达明显低于TNF+HNK组(P<0.05)。结论HNK可能通过激活ERK信号通路引起TRAF3表达上调,进而抑制FLS活化。
Objective To explore the inhibiting effect of honokiol(HNK)on fibroblast-like synoviocyte(FLS)activation and its molecular mechanism.Methods The MH7A cells were treated by TNF-αto establish the FLS activation model,the extracellular signal-regulated kinase(ERK)inhibitor MK-8353 was used to inhibit the ERK activation,and siRNA was used to inhibit TNF-αreceptor associated factor 3(TRAF3)expression.Then the cellular proliferation was detected by CCK-8,the secretion of interleukin(IL)-6 and CXC chemokine ligand 10(CXCL10)was detected by ELISA,the expression of matrix met alloproteinase(MMP)-2,MMP-9,TRAF3 and p-ERK protein was detected by Western blot,and the mRNA expression of TRAF3 was measured by real-time PCR.Results The cellular proliferation in the TNF+HNK group was significantly decreased compared with the group CON and TNF(P<0.05),moreover the excretion of IL-6 and CXCL10 and expressions of MMP-2 and MMP-9 proteins were significantly decreased compared with the group TNF(P<0.05).Compared with the group CON and TNF,the TRAF3 mRNA and protein expression in the group TNF+HNK were significantly increased(P<0.05).The TRAF3 protein expression in the group si-TRAF3 was significantly decreased compared with the group si-CON(P<0.05),the secretion of IL-6 and CXCL10 and expressions of MMP-2 and MMP-9 proteins in the group TNF+HNK+si-TRAF3 were significantly increased compared with the group TNF+HNK(P<0.05).Compared with the group CON and TNF,the p-ERK1/2 protein expression in the group TNF+HNK was significantly increased(P<0.05),the p-ERK1/2 and TRAF3 proteins expression in the group TNF+HNK+MK was significantly lower than that in the group TNF+HNK.Conclusion HNK causes the up-regulation of TRAF3 expression by activating ERK signal,thus inhibits the FLS activation.
作者
吕昌伟
和晶
张乾
郗海涛
强毅
张静涛
LYU Changwei;HE Jing;ZHANG Qian;XI Haitao;QIANG Yi;ZHANG Jingtao(Department of Orthopedics,Affiliated Hospital of Northwestern University/Xi′an Municipal Third Hospital,Xi′an,Shaanxi 710018,China)
出处
《重庆医学》
CAS
2021年第22期3797-3802,共6页
Chongqing medicine
基金
陕西省重点研发计划项目(2019SF-194)。
