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目的:探讨miR-126在截短型rhtBIGH3-(RGD)_2蛋白抑制HUVEC细胞生物学活性中的作用。方法:体外培养VEGF孵化的人脐静脉内皮细胞(VEGF-HUVEC),分别加入rhtBIGH3-(RGD)_2蛋白终浓度为0和100μg/mL,作用24、48、72 h条件下,分别检测Caspase-3活性和miR-126表达水平。在rhtBIGH3-(RGD)_2蛋白终浓度为0和100μg/mL时,分别加入miR-126 mimic和miR-126 inhibitor,作用VEGF-HUVEC 48 h后,Real-time PCR检测miR-126表达水平,通过检测Caspase-3活性来检测细胞凋亡情况。结果:在VEGF-HUVEC中,当rhtBIGH3-(RGD)_2蛋白终浓度为100μg/mL条件下,Caspase-3水平升高,miR-126表达水平升高,在48 h下达到最高峰。在VEGF-HUVEC中,当rhtBIGH3-(RGD)_2蛋白终浓度分别为100μg/mL条件下,加入miR-126mimic后,miR-126表达升高,Caspase-3水平升高;加入miR-126 inhibitor后,48 h后检测miR-126表达下降,Caspase-3水平也下降。结论:截短型rhtBIGH3-(RGD)_2蛋白通过上调miR-126表达从而促进细胞凋亡、抑制HUVEC生物活性,从而抑制角膜新生血管的发生
AIM: To investigate the role of miR-126 in the inhibition of the biological activity of HUVECs by truncated rhtBIGH3- (RGD) _2 protein. Methods: VEGF-HUVECs were cultured in vitro. The final concentration of rhtBIGH3- (RGD) 2 protein was 0 and 100 μg / mL respectively. The cells were treated with VEGF for 24, 48 and 72 h, respectively. The expressions of Caspase- 3 activity and miR-126 expression level. At the final concentrations of rhtBIGH3- (RGD) 2 protein of 0 and 100 μg / mL, miR-126 mimic and miR-126 inhibitor were added respectively and the expression of miR-126 was detected by Real-time PCR Caspase-3 activity was measured to detect apoptosis. Results: In the VEGF-HUVEC, the level of Caspase-3 increased and the expression of miR-126 increased when the final concentration of rhtBIGH3- (RGD) _2 protein was 100 μg / mL, reaching the peak at 48 h. When miR-126mimic was added at the final concentration of rhtBIGH3- (RGD) 2 protein of 100μg / mL in VEGF-HUVEC, the expression of miR-126 increased and the level of Caspase-3 increased. After 48 h, the expression of miR-126 and the level of Caspase-3 also decreased. Conclusion: The truncated rhtBIGH3- (RGD) _2 protein can inhibit the biological activity of HUVEC by up-regulating the expression of miR-126, thereby inhibiting the occurrence of corneal neovascularization