论文部分内容阅读
C42属于epipolythiodioxopiperazines(ETPs)二酮呱嗪类化合物,具有多种生物学活性,包括抑制病毒复制;不过其对hepatitis B virus(HBV)复制的影响及机理鲜有报道。自噬是广泛存在于真核细胞、通过溶酶体降解长半衰期蛋白的现象,参与多种生理、病理过程。有研究发现自噬对HBV的复制至关重要。C42是否通过改变自噬来影响此病毒的复制目前还未见报道。在该研究中,我们发现表达HBV基因组的Hep G2.215细胞较原始的Hep G2细胞,自噬体明显增加并伴随着Akt磷酸化的增高。C42可以降低自噬基因LC3-II和p62的水平,同时会影响Akt信号通路。氯喹是一种自噬抑制剂,它的存在可以抑制C42导致的LC3-II降低,表明C42可以引起该细胞的自噬。敲降自噬基因和抑制Akt磷酸化均可以减少HBV-X蛋白表达。而利用氯喹抑制自噬体与溶酶体的融合却提高了HBV-X蛋白水平。由于HBV-X对该病毒的复制至关重要,因此,我们认为,C42通过自噬和Akt信号通路来抑制HBV的复制。
C42 belongs to the class of epipolythiodioxopiperazines (ETPs), which has a variety of biological activities including inhibition of viral replication; however, its effect on hepatitis B virus (HBV) replication and its mechanisms have rarely been reported. Autophagy is a widespread phenomenon in eukaryotes that degrade long-lived proteins through lysosomes and is involved in a variety of physiological and pathological processes. Some studies have found that autophagy is essential for HBV replication. Whether C42 affects autophagy to affect the replication of this virus has not been reported yet. In this study, we found that Hep G2.215 cells expressing the HBV genome significantly increased autophagosomes and accompanied an increase in Akt phosphorylation compared to the original Hep G2 cells. C42 can reduce the level of autophagy genes LC3-II and p62, while affecting the Akt signaling pathway. Chloroquine is an autophagy inhibitor whose presence inhibits the reduction of LC3-II caused by C42, suggesting that C42 can cause autophagy in this cell. Knockdown of autophagy genes and inhibition of Akt phosphorylation can reduce HBV-X protein expression. The use of chloroquine to inhibit the fusion of autophagosomes and lysosomes increased HBV-X protein levels. Because HBV-X is essential for replication of the virus, we believe that C42 inhibits HBV replication through autophagy and Akt signaling.