【摘要】 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。更多还原

【Abstract】 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.更多还原