绿原酸对颞下颌关节软骨细胞增殖、凋亡和炎症因子释放的影响及作用机制研究

Effects of chlorogenic acid on IL-1β-induced proliferation,apoptosis and inflammatory factor release of temporomandibular joint chondrocytes and its mechanism

目的探讨绿原酸对颞下颌关节骨关节炎的保护作用机制。方法采用10ng/ml的白细胞介素1β(IL-1β)诱导大鼠颞下颌关节软骨细胞损伤,同时使用不同浓度绿原酸对软骨细胞进行干预。采用CCK-8法检测软骨细胞增殖情况,流式细胞术检测细胞凋亡情况,qRT-PCR法检测软骨细胞中肿瘤坏死因子α(TNF-α)、IL-6、基质金属蛋白酶3(MMP-3)、MMP-13、Wnt-5A和Ror2 mRNA的表达水平,Western blot法检测Wnt-5A和Ror2蛋白的表达水平。结果不同浓度绿原酸干预IL-1β诱导的颞下颌关节软骨细胞损伤后,绿原酸能够促进软骨细胞增殖,抑制软骨细胞凋亡,抑制TNF-α、IL-6、MMP-3和MMP-13 mRNA的表达水平,同时抑制Wnt-5A/Ror2信号通路中Wnt-5A和Ror2的表达。结论绿原酸或通过抑制Wnt-5A/Ror2信号通路调控IL-1β诱导损伤的颞下颌关节软骨细胞增殖、凋亡和炎症因子释放。绿原酸或可作为治疗颞下颌关节骨关节炎的药物。

Objective To investigate the effects of chlorogenic acid on IL-1β-induced temporomandibular joint chondrocyte proliferation,apoptosis and inflammatory factor release,and its mechanism.Methods The temporomandibular joint chondrocyte injury was induced by IL-1βat 10ng/ml in rats,and the injured chondrocytes were treated with different concentrations of chlorogenic acid.Chondrocyte proliferation was determined with CCK-8;cell apoptosis was detected with flow cytometry;the mRNA expression levels of NF-α,IL-6,MMP-3,MMP-13,Wnt-5A and Ror2q was detected with RT-PCR;Wnt-5A and Ror2 protein expression levels were detected by Western blot.Results In temporomandibular joint injured by IL-1βchlorogenic acid promoted chondrocyte proliferation,inhibited chondrocyte apoptosis,and down-regulated the mRNA expression of TNF-α,IL-6,MMP-3,MMP-13,and Wnt-5A and Ror2 expression levels in the Wnt-5A/Ror2 signaling pathway.Conclusion Chlorogenic acid can regulate IL-1β-induced chondrocyte proliferation,apoptosis and inflammatory factor release by inhibiting Wnt-5A/Ror2 signaling pathway,indicating that it may be used to treat temporomandibular joint osteoarthritis.