关键词: 人巨细胞病毒, 氧化应激, 细胞自噬, 人包皮成纤维细胞

Abstract: Objective To provide new insights into the prevention and treatment of human cytomegalovirus (HCMV) by observing the effect of autophagy induced by oxidative stress on the proliferation of HCMV in cells. Methods Using PCR to amplify the LC3B encoding fragment, an autophagy expression vector was constructed to observe the formation of cellular autophagy. Then, HFF cells infected with HCMV (AD169) for 72 hours were collected. The levels of UL122 expression and viral particles were detected by real-time quantitative PCR. And the viral protein pp65 expression level and the proliferation of virion were detected by Western blot and TCID₅₀, respectively. Results Autophagy detection vectors were successfully constructed, which could be used to indicate the formation of cellular autophagy. Compared with normal cultured cells, oxidative stress could induce autophagy formation, and upregulated UL122 gene expression, pp65 protein levels and viral load by this pathway. The viral titer test also showed that autophagy could promote the replication of HCMV in cells. Conclusion Oxidative stress can induce autophagy and promote the replication and proliferation of HCMV, while autophagy inhibitor 3-MA can inhibit the replication of HCMV promoted by oxidative stress. It is confirmed that autophagy is one of the mechanisms of oxidative stress promoting the replication and proliferation of HCMV.

Key words: Human cytomegalovirus, Oxidative stress, Autophagy, Human foreskin fibroblasts