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 |
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引用本文: | 宋凡倩,高宁宁,孙莉尧,曹志超,刘平,葛红岩.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[J].现代生物医学进展,2017,17(2):225-228. |
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作者姓名: | 宋凡倩 高宁宁 孙莉尧 曹志超 刘平 葛红岩 |
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作者单位: | 哈尔滨医科大学第一临床学院眼科医院;哈尔滨医科大学 |
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基金项目: | 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) |
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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,从而抑制角膜新生血管的发生
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关 键 词: | rhtBIGH3-(RGD)2蛋白 miR-126 HUVEC 角膜新生血管 Caspase-3 |
Biological Activities of HUVECS Is Inhibited by Truncated rhtBIGH3-
(RGD)2 Protein though the Up-regulation of miR-126 |
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Abstract: | 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 ug/mL and 100
ug/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 ug/mL and 100 ug/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 ug/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 ug/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.Conclusion:The truncated rhtBIGH3- (RGD)2 protein promote apoptosis and inhibit HUVEC activity by increasing miR-126
expression, thus inhibiting corneal neovascularization. |
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Keywords: | RhtBIGH3-(RGD)2 protein MiR-126 HUVEC Corneal neovascularization Caspase-3 |
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