自噬相关基因Atg7的表达对乙型肝炎病毒复制的影响

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摘要目的探讨Atg7的表达对乙型肝炎病毒感染肝细胞中HBV复制的影响。方法选取2019年4～5月首都医科大学附属北京市佑安医院慢性乙型肝炎病毒感染者的肝组织石蜡切片，采用免疫荧光染色观察Atg7、LC3和HBs的表达情况；利用Atg7表达质粒和小干扰RNA在HepG2.2.15细胞中建立Atg7过表达组及Atg7减少组，给予不同时间段的饥饿处理，利用蛋白质免疫印迹和实时定量PCR观察两组模型中自噬及HBV病毒复制水平的变化。采用SPSS 17.0统计学软件进行数据分析。结果在HBs阳性细胞中，Atg7和LC3的表达量高于HBs阴性细胞；Atg7过表达饥饿处理后，自噬水平、HBc和HBV-DNA表达量高于对照，且自噬和HBV的复制水平在饥饿6 h增加最显著，差异有高度统计学意义（P < 0.01）。Atg7敲减饥饿处理后，细胞自噬水平、HBc和HBV DNA表达量均低于对照，在饥饿6 h减少最为显著，差异有高度统计学意义（P < 0.01）。结论 HBV感染引起Atg7的表达增加，自噬增强；在自噬早期干预Atg7的表达会显著影响HBV复制。

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	王阳车阳高玉雪杨通旺张玉林陈德喜石英

关键词 ：自噬相关基因Atg7, 乙型肝炎病毒, 自噬, 乙肝病毒脱氧核糖核酸

Abstract：Objective To investigate the effect of Atg7 expression on HBV replication in hepatocytes infected with hepatitis B virus. Methods The expression of Atg7, LC3 and HBs in paraffin section of liver tissue of patients with chronic hepatitis B virus infection in Beijing You′an Hospital from April to May 2019 was observed by immunofluorescence staining. Atg7 overexpression group and Atg7 reduction group were established in HepG2.2.15 cells by using Atg7 expression plasmids and small interfering RNAs. The changes of autophagy and HBV replication level were observed by Western blot and real-time quantitative PCR. Data was analyzed by SPSS 17.0. Results The expression of Atg7 and LC3 in HBs positive cell was higher than that in HBs negative cell; after Atg7 overexpression and starvation, autophagy level, HBc and HBV-DNA expression were higher than those of control group, the replication level of autophagy and HBV increased most significantly in 6 h of starvation, and the differences were highly statistically significant (P < 0.01). The level of autophagy, the expression of HBc and HBV DNA in Atg7 treated cells were lower than those in control group, the decrease was most significant at 6 h after starvation, and the differences were highly statistically significant (P < 0.01). Conclusion HBV infection increases Atg7 expression and autophagy, early intervention of Atg7 expression in autophagy can significantly affect HBV replication.

Key words： Autophagy related gene Atg7 HBV Autophagy HBV-DNA

基金资助:国家自然科学基金资助项目（81470098）；
北京市自然科学基金资助项目（7182072）。

通讯作者: 石英（1976.12-），女，博士，副教授；研究方向：生物化学与分子生物学。

作者简介: 王阳（1991.5-），女，首都医科大学2017级生物化学与分子生物学专业在读硕士研究生；研究方向：病毒性肝炎及肝癌。

引用本文:

王阳车阳高玉雪杨通旺张玉林陈德喜石英. 自噬相关基因Atg7的表达对乙型肝炎病毒复制的影响[J]. 中国医药导报, 2020, 17(2): 4-8.
WANG Yang CHE Yang GAO Yuxue YANG Tongwang ZHANG Yulin CHEN Dexi SHI Ying. Effect of expression of autophagy related gene Atg7 on replication of hepatitis B virus. 中国医药导报, 2020, 17(2): 4-8.

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http://www.yiyaodaobao.com.cn/CN/ 或 http://www.yiyaodaobao.com.cn/CN/Y2020/V17/I2/4

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