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

作　　者：宋凡倩 高宁宁[1] 孙莉尧 曹志超[2] 刘平[1] 葛红岩[1] 

机构地区：[1]哈尔滨医科大学第一临床学院眼科医院,黑龙江哈尔滨150001 [2]哈尔滨医科大学,黑龙江哈尔滨150001

出　　处：《现代生物医学进展》2017年第2期225-228,共4页Progress in Modern Biomedicine

基　　金：黑龙江省教育厅面上项目(12541516);国家自然科学青年基金项目(81300728)

摘　　要：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.

关 键 词：rhtBIGH3-(RGD)2蛋白 MIR-126 HUVEC 角膜新生血管 Caspase-3 

分 类 号：R772.23[医药卫生—眼科]