Effect of 27nt-miRNA on apoptosis of human umbilical vein endothelial cells induced by oxidized low-density lipoprotein |
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Authors: | ZHAO Yan-jiao YAN Yuan-yuan LI Dan LUO Xue-lan YANG Peng ZHOU Fu-long YANG Rui-xia TANG Shuang-yi OU He-sheng |
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Institution: | 1.Pharmaceutical College of Guangxi Medical University, Nanning 530021, China;2.Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530201, China;3.Department of Pharmacy, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China. |
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Abstract: | AIM To investigate the effect of 27nt-miRNA (27nt-miR) on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by oxidized low-density lipoprotein (Ox-LDL) and its underlying mechanism. METHODS HUVECs were cultured in vitro and grouped as below: normal control group, Ox-LDL group, 27nt-miR+Ox-LDL group, anti-27nt-miR+Ox-LDL group and negative control+Ox-LDL group. The cells in Ox-LDL group were treated with Ox-LDL at 40 mg/L for 48 h, while those in normal control group were untreated but cultured normally. The cells in 27nt-miR+Ox-LDL group, anti-27nt-miR+Ox-LDL group and negative control+Ox-LDL group were transfected with their corresponding lentiviral vectors under the same procedure, followed by treatment with Ox-LDL at 40 mg/L for 48 h to induce apoptosis. The cell viability was measured by CCK-8 assay. The migration capacity was detected by scratch assay. The caspase-3 activity was measured by caspase-3 activity assay kit. The apoptotic rate was analyzed by Hoechst 33258 and flow cytometry. The mRNA and protein expression levels of Bcl-2, Bax and caspase-3 were determined by RT-qPCR and Western blot. RESUITS: Compared with negative control+Ox-LDL group, the cell viability and migration ability were significantly decreased by over-expression of 27nt-miR in the HUVECs (P<0.05), while the activity of caspase-3 and apoptosis induced by Ox-LDL were significantly increased (P<0.05). Furthermore, the mRNA and protein expression levels of Bax and caspase-3 were significantly up-regulated (P<0.05), and the mRNA and protein expression level of Bcl-2 was down-regulated in 27nt-miR+Ox-LDL group (P<0.05). Meanwhile, all the above indexes showed an opposite tendency in anti-27nt-miR+Ox-LDL group. CONCLUSION 27nt-miR promotes Ox-LDL-induced apoptosis and inhibits the viability and migration of HUVECs in vitro, possibly through regulating the expression of apoptotic/anti-apoptotic proteins such as Bax,caspase-3 and Bcl-2. |
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Keywords: | 27nt-miRNA Human umbilical vein endothelial cells Oxidized low-density lipoprotein Apoptosis |
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