桂皮醛经JAK2/STAT3通路抑制VEGF诱导的内皮细胞增殖、迁移及成管

Inhibitory Effect of Cinnamaldehyde on Proliferation,Migration and Tube Formation of VEGF-induced Endothelial Cells via JAK2/STAT3 Pathway

目的:为探讨桂皮醛对糖尿病视网膜病变新生血管的作用及机制,观察了桂皮醛对血管内皮生长因子(VEGF)诱导EA. hy926细胞增殖、迁移、成管以及Janus激酶2/信号传导与转录激活因子3(JAK2/STAT3)通路的影响。方法:将EA.hy926细胞分成空白组、模型组(7μg·L-1VEGF),VEGF+桂皮醛(60,90,120,150μmol·L-1)组,分别采用噻唑蓝(MTT)比色法和划痕实验检测桂皮醛对VEGF诱导EA. hy 926细胞增殖和迁移作用的影响;将EA. hy 926细胞分成空白组,模型组(7μg·L-1VEGF),VEGF+桂皮醛(90,150μmol·L-1)组,采用管腔形成实验检测桂皮醛对VEGF诱导EA. hy 926细胞成管作用的影响;将EA. hy 926细胞分成空白组,模型组(7μg·L-1VEGF),VEGF+AG490 (50μmol·L-1)组,VEGF+桂皮醛(90μmol·L-1)组,VEGF+桂皮醛(150μmol·L-1)组,VEGF+桂皮醛(150μmol·L-1)+AG490(50μmol·L-1)组,采用蛋白免疫印迹法(Western blot)检测桂皮醛对VEGF诱导EA. hy 926细胞JAK2/STAT3通路的影响。结果:与空白组比较,模型组能够显著地促进EA. hy 926细胞增殖和迁移(P <0. 01)。与模型组比较,桂皮醛(60,90,120,150μmol·L-1)组能显著抑制VEGF诱导EA. hy 926细胞的增殖和迁移(P <0. 01)。与空白组比较,VEGF对EA. hy 926细胞成管具有一定的促进作用,成管的节点数、交叉点数、网眼数和血管分支数均有增加,但无统计学差异。与模型组比较,桂皮醛(90,150μmol·L-1)组对成管的节点数、交叉点数和网眼数均有明显抑制作用(P <0. 05,P <0. 01)。与空白组比较,模型组p-JAK2,p-STAT3,STAT3蛋白表达明显升高(P <0. 05,P <0. 01)。与模型组比较,桂皮醛(150μmol·L-1)能够显著抑制VEGF引起的p-JAK2,p-STAT3,STAT3蛋白表达升高(P <0. 01),桂皮醛(90μmol·L-1)能够显著抑制VEGF引起的p-STAT3,STAT3蛋白表达升高(P <0. 05,P <0. 01)。结论:桂皮醛对VEGF诱导EA. hy 926细胞的增殖、迁移、成管具有明显的抑制作用,该作用与抑制JAK2/STAT3通路的激活有关。

Objective: To investigate the effect and mechanism of cinnamaldehyde on the angiogenesis of diabetic retinopathy,and the effect of cinnamaldehyde on vascular endothelial growth factor( VEGF) induced proliferation,migration,tube formation and Janus kinase 2/signal transducer and activator of transcription 3( JAK2/STAT3) pathway of EA. hy 926 cells were observed. Method: EA. hy 926 cells were divided into normal control group,model group( 7 μg·L-1 VEGF),and VEGF + cinnamaldehyde group( 60,90,120,150 μmol·L-1).The methyl thiazolyl tetrazolium( MTT) assay and scratch test were used to observe the effect of cinnamaldehyde on the proliferation and migration of EA. hy 926 cells induced by VEGF. EA. hy 926 cells were divided into normal control group,model group( 7 μg·L-1 VEGF),and VEGF + cinnamaldehyde group( 90,150 μmol·L-1). The tube formation experiment was used to observe the effect of cinnamaldehyde on the tube formation of EA. hy 926 cells induced by VEGF. EA. hy 926 cells were divided into normal control group,model group( 7 μg·L-1 VEGF),VEGF + AG490 group( 50 μmol·L-1),VEGF + cinnamaldehyde group( 90 μmol·L-1),VEGF + cinnamaldehyde group( 150 μmol·L-1),and VEGF + cinnamaldehyde group( 150 μmol·L-1) + AG490 group( 50 μmol·L-1).Western Blot method was used to explore the effect of cinnamaldehyde on the JAK2/STAT3 signaling pathway in EA. hy 926 cells induced by VEGF. Result: Compared with the control group,model group obviously promoted the proliferation and migration of EA. hy 926 cells( P < 0. 01). Compared with the model group,cinnamaldehyde( 60,90,120,150 μmol·L-1) significantly suppressed VEGF-induced proliferation and migration of EA. hy 926 cells( P < 0. 01). Compared with the control group,VEGF group could promote the tube formation of EA. hy 926 cells. The number of nodes,junctions,meshes and vascular branches were increased,but with no statistical difference. Compared with the model group,cinnamaldehyde( 90,150 μmol·L-1) showed an obvious inhibitory effect on the number of nodes,junctions and meshes of tubules( P < 0. 05,P < 0. 01). Compared with the control group,the expressions of p-JAK2,p-STAT3,and STAT3 in the model group were significantly increased( P <0. 05,P < 0. 01). Compared with the model group,Cinnamaldehyde( 150 μmol·L-1) significantly reduced the expressions of P-JAK2,P-STAT3,STAT3 proteins( P < 0. 01). Cinnamaldehyde( 90 μmol·L-1) obviously reduced the expressions of p-STAT3 and STAT3 proteins( P < 0. 05,P < 0. 01). Conclusion: Cinnamaldehyde showed a significantly inhibitory effect on the proliferation,migration and tube formation of VEGF-induced EA. hy 926 cells,which was related to the inhibition of the activation of JAK2/STAT3 pathway.