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目的:探讨中国圆田螺多糖(PCC)体外抗乙肝病毒的生物学活性。方法:以HepG2 2.2.15细胞系为体外实验模型。MTT法检测细胞毒性,将不同安全浓度的PCC及阳性对照药物3TC作用于此细胞系,实验同时设不加药物的细胞对照,第9天收集细胞培养上清液。采用ELISA法测定各组细胞培养上清液中HBsAg,HBeAg水平,荧光探针定量PCR检测HBV-DNA含量。结果:PCC在HepG2 2.2.15细胞培养中的TC0为1 g·L-1,TC50>10 g·L-1,对细胞毒性较小。PCC对HepG2 2.2.15细胞HBsAg,HBeAg分泌的IC50分别为0.501,0.401 g·L-1,SI分别为>19.96和>24.94。PCC可以有效抑制HBsAg,HBeAg的分泌,且PCC对HBeAg的抑制效果优于HBsAg。PCC在0.1,1 g·L-1(P<0.05)对HepG2 2.2.15细胞中的HBV-DNA有明显的抑制作用。结论:中国圆田螺多糖具有一定的体外抗HBV活性,且毒性较小,具有良好的应用前景。
Objective: To investigate the biological activity of Chinese medicinal polysaccharide (PCC) against hepatitis B virus in vitro. Methods: The HepG2 2.2.15 cell line was used as an experimental model in vitro. MTT assay cytotoxicity, different concentrations of safe PCC and positive control drug 3TC role in this cell line, the experiment also set the drug-free cell control, the collection of cell culture supernatant on the 9th day. The HBsAg and HBeAg levels in the cell culture supernatants of each group were measured by ELISA, and the content of HBV-DNA was detected by fluorescent probe quantitative PCR. Results: The TC0 of PCC in HepG2 2.2.15 cell culture was 1 g · L-1 and TC50> 10 g · L-1, which showed little cytotoxicity. The IC50 of HBsAg and HBeAg secreted by PCC in HepG2 2.2.15 cells were 0.501 and 0.401 g · L -1, respectively, and SI> 19.96 and> 24.94, respectively. PCC can effectively inhibit the secretion of HBsAg and HBeAg, and the inhibitory effect of PCC on HBeAg is better than that of HBsAg. PCC at 0.1,1 g · L-1 (P <0.05) significantly inhibited HBV-DNA in HepG2 2.2.15 cells. CONCLUSION: Polysaccharide from China has a certain degree of anti-HBV activity in vitro with less toxicity and good application prospect.