Research The Autophagy In The Changes Of Vascular Endothelial Cell Biological Behavior Caused By Human Cytomegalovirus Infection

Objective In this study, the autophagy regulation drugs were used to investigate the role of autophagy in the biological behavior changes of vascular endothelial cells caused by human cytomegalovirus(HCMV) infection. The purpose of this study was to understand the role of autophagy and mTOR signaling pathways in the atherosclerosis progression induced by HCMV infection.Method By using autophagy promoter LiCl, autophagy inhibitors 3-MA, mTOR signaling pathways inhibitor Rapamycin to deal with the HUVEC cells after HCMV infection. By using autophagy fluorescence staining and pGFP-LC3 plasmid to detect the phenomenon of cell autophagy. By using MTT and Transwell assay to detect HUVEC proliferation and migration. Also, by using western boltting,the expression levels of the LC3 protein and mTOR signal pathway protein p70s6k-p were detected. By using Enzyme-linked immuno sorbent assay, the expression level of HUVEC cell adhesion factor were detected. By using real time qPCR, the HCMV copies in HUVEC, that were treated by autophagy modulation, were detectedResults By AO and MDC staining, tranfecting HUVECs with GFP-LC3, it was found that in HCMV-infected cells, an increasing number of punctuate structures per cell from 24h to 48h p.i. while a clear decreasing number at 60h p.i. these results suggest that autophagy was induced early in response to virus infection,,however it was quickly transformed by the virus at the time of 48h p.i. By using mTOR autophagy pathway inhibitor rapamycin to inhibit mTOR pathway of autophagy, it was found that the influences of mTOR signaling pathways to HCMV propagation in HUVEC cells existed. By using real time fluorescent qPCR, rapamycin was found that it could in some extent promote HCMV proliferation in HUVEC cells. In subsequent experiments, the role of autophagy in the changes of the behavior of vascular endothelial cells treated with HCMV infection were also analyzed. After HCMV continued to infected HUVEC, the detection of invasion Assay revealed that HCMV infection could cause HUVEC cell migration, and the detection of ELISA assay revealed that the expression level of Adhesion molecule of HUVEC cells increased after HCMV infection.Conclusions In vitro infection model of HUVEC by HCMV AD 169 strain was successfully established. Through regulating the autophagy and mTOR signaling pathways, it could change the autophagy in HUVEC cells and virus replication, as well as the inhibition of the biological behavior change of HUVEC cell that was infected by HCMV. These results could prompt that the intervention of mTOR autophagy signaling pathways can be used as an effective method for treatment of atherosclerosis.