截短型rhtBIGH3-(RGD)_2蛋白通过上调miR-126表达抑制人脐静脉内皮细胞的生物活性

Biological Activities of HUVECS Is Inhibited by Truncated rhtBIGH3-(RGD)_2 Protein though the Up-regulation of miR-126

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。

Objective： To explore the effect of truncated rhtBIGH3 protein in inhibiting the biological activities of HUVECS.Methods： Human umbilical vein endothelial cells（HUVECS） were cultured with VEGF in vitro, supplemented with 0 μg/mL and 100μg/mL rhtBIGH3-（RGD）_2 protein. HUVECS were incubated for 24 h, 48 h, 72 h, and testing Caspase-3 activities and miR-126 expression level. Added miR-126 mimic and miR-126 inhibitor when the rhtBIGH3-（RGD）_2 concentration was 0 μg/mL and 100 μg/mL,after 48 h, use Real-time PCR testing the miR-126 expression level, and test the apoptosis though the activity of Caspase-3. Results： In the VEGF-HUVEC supplemented with 100 μg/mL rhtBIGH3-（RGD）_2 protein, the expression of both Caspase-3 and miR-126 raised, and reach the peak at 48 h. In VEGF-HUVEC, when the final concentration of rhtBIGH3-（RGD）_2 protein was 100 μg/mL, after the addition of mimic miR-126, miR-126 and Caspase-3 expression increased, after adding inhibitor 48 h, miR-126 and Caspase-3 expression decreased. Conclusions： The truncated rhtBIGH3-（RGD）_2 protein promote apoptosis and inhibit HUVEC activity by increasing miR-126 expression, thus inhibiting corneal neovascularization.