Chemerin/CMKLR1激活自噬和促进体外视网膜新生血管形成

Chemerin/CMKLR1 Activates Autophagy and Promotes Retinal Neovascularization in vitro

目的研究Chemerin/CMKLR1对RF/6A细胞自噬和血管形成的影响。方法将RF/6A细胞分为对照组,1、10、100 nmol/L chemerin组,CMKLR1受体抑制剂组(10 nmol/L chemerin+α-NETA)。培养24 h后利用Western印迹检测自噬相关蛋白LC3、Beclin-1表达水平,CCK8法、Transwell及基质胶法检测RF/6A细胞增殖,迁移,管腔形成。结果LC3、Beclin-1在3种浓度chemerin组表达水平较对照组高(P<0.05),与10 nmol/L chemerin组相比,CMKLR1受体抑制剂组LC3、Beclin-1表达水平降低(P<0.05)。相比对照组,10 nmol/L、100 nmol/L chemerin组促进细胞增殖,而抑制剂组作用相反(P<0.05);对照组、10 nmol/L chemerin组、CMKLR1受体抑制剂组3组细胞迁移数分别是52.8±5.6、69.5±3.6、39.5±7.5(P<0.05),3组细胞管腔形成长度分别是10215±912、12925±1768、10672±735(P<0.05),差异具有统计学意义。结论Chemerin/CMKLR1通路能够激活RF/6A细胞自噬,促进细胞增殖、迁移和管腔形成。

Objective To investigate the effects of Chemerin/CMKLR1 on autophagy and angiogenesis in RF/6A cells.Methods RF/6A cells were divided into control group,chemerin group with three concentrations(1,10,100 nmol/L),CMKLR1 receptor inhibitor group(10 nmol/L chemerin+α-NETA).After 24 h of culture,the expression levels of autophagy related proteins LC3 and beclin-1 were detected by Western blotting.The proliferation,migration,tube formation of RF/6A cells were detected by using CCK8 assay,Transwell assay,and matrigel assay,respectively.Results The expression levels of LC3 and Beclin-1 in the chemerin group were higher than those in the control group(P<0.05),and the expression levels of LC3 and Beclin-1 in the CMKLR1 receptor inhibitor group were lower than those in the 10 nmol/L chemerin group(P<0.05).Compared with the control group,chemerin at 10 nmol/L and 100 nmol/L concentration could promote cell proliferation,CMKLR1 receptor inhibitors had the opposite effect(P<0.05).The cell migration numbers of control group,10 nmol/L chemerin group and CMKLR1 receptor inhibitor group were 52.8±5.6,69.5±3.6 and 39.5±7.5(P<0.05),respectively.The length of tube formation was 10215±912,12925±1768 and 10672±735 in the three groups(P<0.05),and the difference was statistically significant.Conclusion Chemerin/CMKLR1 pathway can activate RF/6A cell autophagy and promote cell proliferation,migration and tube formation.