全蝎和蜈蚣的多肽粗提物对肝癌HepAD38细胞中HBV的抑制作用

目的: 研究全蝎尾部多肽粗提物（peptide extract from scorpion tail，PEST）与蜈蚣头部多肽粗提物（peptide extract from centipede head, PECH）对肝癌细胞乙型肝炎病毒（Hepatitis B virus, HBV）合成的影响及其可能机制。方法：超滤法提取PEST与PECH。选取肝癌HepAD38和HepG2细胞，将其分组：对照组，用含10%胎牛血清的培养基培养；PEST组，不同浓度PEST（0.125、0.250、0.500和1.000 mg/mL）处理；PECH组，不同浓度PECH（0.125、0.250、0.500和1.000 mg/mL）处理；阳性对照组，拉米夫定或恩替卡韦或四环素处理。MTT法检测细胞活力，qRT PCR检测HepAD38细胞上清液HBV DNA含量。另取HepAD38细胞，分为PEST组（1 mg/mL PEST处理）、PECH组（1 mg/mL PECH处理）、对照组、阳性对照组和阴性对照组；ELISA法检测细胞上清液中HBsAg和HBeAg含量，qRT PCR检测HBV X、S、preC、P mRNA表达量，蛋白印迹法检测HBV核心蛋白（HBV core protein，HBc）表达。结果：PEST或PECH处理的肝癌细胞HepG2和HepAD38细胞活力均在70%以上；与对照组相比，PEST组（0.250、0.500和1.000 mg/mL）和PECH组（0.125、0.500和1.000 mg/mL）HBV DNA拷贝数明显降低（P均<0.05）；与对照组相比，PEST组和PECH组HBsAg和HBeAg含量明显降低（P均<0.05），HBV P mRNA相对表达明显降低（P均<0.05），PEST组HBc表达几乎无差异，而PECH组HBc表达减少。结论：PEST和PECH在HepAD38细胞株中表现出抗HBV作用，可能与其抑制HBV P mRNA合成有关。

Inhibition of Hepatitis B virus in liver cancer cells by preliminary peptide extracts of scorpion and centipede

Objective: To explore the effects of scorpion peptide preliminary extract （peptide extract from scorpion tail, PEST） and the centipede preliminary peptide extract （peptide extract from centipede head, PECH） on Hepatitis B virus synthesis and potential mechanism in liver cancer cells. Methods: Ultrafiltration was used to extract PEST and PECH. HepAD38 and HepG2 cells were selected and divided into control group, cells were cultured with high sugar medium containing with 10% fetal bovine serum; PEST group, cells were treated with different concentrations （0.125, 0.250, 0.500 and 1.000 mg/mL） of PEST; PECH group, cells were treated with different concentrations （0.125, 0.250, 0.500，1.000 mg/mL） of PECH; positive control group, cells were treated with lamivudine or entecavir or tetracycline; MTT assay was used to detect cell viability. The qRT PCR was used to detect Hepatitis B virus DNA content in the supernatant of HepAD38 cells. In addition, HepAD38 cells were divided into PEST group （cells were treated with 1 mg/mL PEST）, PECH group （cells were treated with 1 mg/mL PECH）, control group, positive control group and negative group; HBsAg and HBeAg content were detected by ELISA; HBV X，S，preC，P mRNA relative expression level were detected by qRT PCR; Hepatitis B virus core protein synthesis was detected by Western blotting. Results: The cell viability of HepG2 and HepAD38 cells treated with PEST or PECH were both above 70%；compared with the control group, PEST（0.250, 0.500 and 1.000 mg/mL） or PECH （0.125, 0.500 and 1.000 mg/mL） group showed significantly decreased Hepatitis B virus DNA copy number （all P<0.05）; compared with the control group, PEST or PECH group showed significantly lower content of HBsAg and HBeAg and expression level of Hepatitis B virus P mRNA （all P<0.05）, PEST group had almost no effect on core protein expression, while PECH group decreased Hepatitis B virus core protein expression. Conclusion: PEST and PECH exerted anti Hepatitis B virus effect on HepAD38 cell, which may be related to its inhibition on P mRNA synthesis of Hepatitis B virus.