Chemerin/CMKLR1激活自噬促进ARPE-19细胞炎症因子表达

Chemerin/CMKLR1 activates autophagy and promotes expression of inflammatory cytokines in ARPE-19 cells

目的研究趋化素/趋化因子样受体1(Chemerin/Chemokine-like receptor 1)对人视网膜色素上皮细胞ARPE-19自噬和炎症因子表达的影响。方法将ARPE-19细胞分为7组:空白对照组、1 nmol/L chemerin组、10 nmol/L chemerin组、100 nmol/L chemerin组、1 nmol/L chemerin+α-NETA组、10 nmol/L chemerin+α-NETA组、100 nmol/L chemerin+α-NETA组,采用CCK8法检测细胞增殖;将细胞分为5组:空白对照组、1 nmol/L chemerin组、10 nmol/L chemerin组、100 nmol/L chemerin组、10 nmol/L chemerin+α-NETA组,采用Western blot检测自噬相关蛋白LC3和Beclin-1表达。将细胞分为3组:空白对照组、10 nmol/L chemerin组、10 nmol/L chemerin+α-NETA组,使用Transwell检测细胞迁移,Western blot检测IL-1β、TNF-α蛋白表达水平。结果与空白对照组相比,1 nmol/L chemerin组、10 nmol/L chemerin组、100 nmol/L chemerin组细胞增殖水平升高(P<0.05);与相应浓度的chemerin组比较,加入α-NETA后细胞增殖均受到抑制(P<0.05)。与空白对照组相比,1 nmol/L chemerin组、10 nmol/L chemerin组、100 nmol/L chemerin组LC3、Beclin-1蛋白表达水平升高(P<0.05);与10 nmol/L chemerin组相比,10 nmol/L chemerin+α-NETA组LC3、Beclin-1蛋白表达水平降低(P<0.05)。与空白对照组相比,10 nmol/L chemerin组细胞迁移数增加,IL-1β、TNF-α蛋白表达水平升高,差异具有统计学意义(P<0.05);与10 nmol/L chemerin组相比,10 nmol/L chemerin+α-NETA组细胞迁移数减少,IL-1β、TNF-α蛋白表达水平降低(均P<0.05)。结论Chemerin/CMKLR1能够激活ARPE-19自噬,促进细胞增殖和迁移,促进炎症因子IL-1β、TNF-α的表达水平。

Objective To investigate the effects of Chemerin/chemokine-like receptor 1(CMKLR1)on the expression of autophagy and inflammatory factors in human retinal pigment epithelial cells(ARPE-19).Methods ARPE-19 cells were divided into seven groups:blank control group,1 nmol/L chemerin group,10 nmol/L chemerin group,100 nmol/L chemerin group,1 nmol/L chemerin+α-NETA group,10 nmol/L chemerin group chemerin+α-NETA group,100 nmol/L chemerin+α-NETA group,and then cell proliferation was detected by CCK8 method.ARPE-19 cells were divided into five groups:blank control group,1 nmol/L chemerin group,10 nmol/L chemerin group,100 nmol/L chemerin group,10 nmol/L chemerin+α-NETA group,and the expression levels of auto-phagy-related proteins LC3 and Beclin-1 were detected by Western blot.ARPE-19 cells were divided into three groups:blank control group,10 nmol/L chemerin group and 10 nmol/L chemerin+α-NETA group,and then the cell migration was detected by Transwell,and the expression levels of IL-1βand TNF-αwere detected by Western blot.Results Compared with blank control group,the cell proliferation was increased in 1 nmol/L chemerin group,10 nmol/L chemerin group and 100 nmol/L chemerin group(P<0.05).Compared with the corresponding chemerin groups,the cell proliferation was inhibited after NETA in chemerin+α-NETA groups(P<0.05).Compared with blank control group,the protein expression levels of LC3 and Beclin-1 in 1 nmol/L chemerin group,10 nmol/L chemerin group and 100 nmol/L chemerin group were increased(P<0.05).Compared with 10 nmol/L chemerin group,the protein expression levels of LC3 and Beclin-1 in 10 nmol/L chemerin+α-NETA group were decreased(P<0.05).Compared with blank control group,the number of cell migration and the protein expression levels of IL-1βand TNF-αwere increased in 10 nmol/L chemerin group(P<0.05).Compared with 10 nmol/L chemerin group,the cell migration number,and IL-1βand TNF-αprotein expression levels were reduced in 10 nmol/L chemerin+α-NETA group(P<0.05).Conclusion Chemerin/CMKLR1 can activate ARPE-19 autophagy,promote cell proliferation and migration,and promote the expression levels of inflammatory cytokines IL-1βand TNF-α.