[Study of the inhibitory effect of cell entry inhibitory myrcludex-B synthetic peptide on HepaRG cells infected with hepatitis B virus in an in vitro infection model]

[体外感染模型中细胞进入抑制肽myrcludex-B对乙型肝炎病毒感染的HepaRG细胞的抑制作用研究]

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Abstract

Objective: To establish and explore the inhibitory effect of cell entry inhibitor myrcludex-B on human liver cancer cell (HepaRG) infected with hepatitis B virus (HBV) positive serum in an in vitro infection model. Methods: HepaRG induced mature cells were divided into polyethylene glycol (PEG) infection group and non-PEG infection group, and then the two infection groups were divided into treatment group with myrcludex-B peptide fragment and control group without myrcludex-B peptide fragment, and the cells were infected with the serum of HBV DNA-positive patients. The expression of HBV DNA in supernatant was detected by fluorescence quantitative PCR, and the HBsAg and HBeAg content in supernatant were detected by chemical fluorescence. Measurement data of the non-normal distribution between the two groups was determined by rank-sum test, and normal distribution was determined by t-test. P < 0.05 was considered as statistically significant. Results: In the treatment and control group the virus titers of the PEG infection group and the non-PEG infection group were (103 877.00 ± 49 013.24) copies / ml and (5 050.09 ± 631.26) copies/ml, and (116 188.57 ± 50 957.19) copies/ml, (5 119.34 ± 1 102.43) copies/ml, respectively. Virus titers of both groups with PEG infection were significantly higher than non-PEG infection group, and the difference was statistically significant (P < 0.05). In the PEG infection group, the virus titers of the peptide-treated group and the non-peptide control group were (103 877.34 ± 49 013.24) copies /ml and (116 188.57 ± 50 957.19) copies/ml, respectively, and the virus titers of the peptide-treated group was decreased significantly (P < 0.05), and the difference was statistically significant. HBsAg content in the supernatant of this group was positive at 0, 1, 2 and 3 days, while the 0 day HBsAg supernatant treated with peptide fragment was significantly lower than that of the control group without peptide fragment. Conclusion: It is feasible to infect HepaRG cells with HBV positive serum supplemented with PEG in vitro, and the cell entry inhibitor myrcludex-B has a certain inhibitory effect on hepatitis B virus infection in this experimental model.

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