目的 在乙型肝炎病毒(HBV)基因转染的人肝癌细胞系(Hep G2)的2215细胞培养中,观察芹灵冲剂(Qinling Chongji,QL)对细胞的毒性和对乙型肝炎病毒表面抗原(HBsAg)和e抗原(HBeAg)的抑制效果。 方法 ①2215细胞培养:将配制成细胞(1×10~5个/mL)接种细胞培养板,96孔板每孔0.2mL,24孔板每孔1mL,37℃50mL/L CO_2培养24h,细胞长成单层后进行实验。②药物对细胞毒性试验:QL 12g/L用培养液2倍稀释成6个浓度:8,4,2,1,0.5g/L,加入96孔细胞培养板,每浓度4孔,每4d换同浓度药液,培养12d,观察药物对细胞的毒性,设无药物细胞对照。③药物对HBsAg,HBeAg抑制试验:在无毒浓度以下2倍稀释试验药液,3个稀释度分别为4,2,1,0.5g/L,每浓度3孔,37℃ 50mL/L CO_2培养,每4d收取培养液,换原浓度药液培养,将d8,d12收集的培养液同时测定HBsAg和HBeAg。④HBsAg和HBeAg测定:用固相放射免疫测定盒测定,方法见说明书,用γ-计数器测定每孔药液cpm值。 结果 QL 4g/L加入细胞培养8d和12d,三批实验对细胞的半数中毒浓度(TC50)平均为8.7g/L±0.3g/L,最大无毒浓度(TC0))为4g/L。在最大无毒浓度(4g/L)时对细胞分泌HBeAg平均抑制率(8d)为51.1％±2％和(12d)为65.0％±1.9％,半数有效剂量(IC_(50))平均(8d)为3.1g/L±0.8g/L和(12d)为9.1g/L±0.9g/L,与 Objective To observe the toxicity of Qinling Chongji (QL) on cells and the surface antigen of hepatitis B virus in 2215 cell culture of human hepatoma cell line (Hep G2) transfected with hepatitis B virus (HBV) gene ( Inhibitory effect of HBsAg) and e antigen (HBeAg). Method 12215 Cell Culture: Prepare cells (1×10~5 cells/mL) into cell culture plates, 0.2 mL per well in 96-well plates, 1 mL per well in 24 well plates, and culture at 37°C for 50 hours at 50 mL/L CO 2 for 24 hours. After a single layer experiment. 2 Drugs for cytotoxicity test: QL 12g/L was diluted 2-fold with culture medium into 6 concentrations: 8, 4, 2, 1, 0.5 g/L, and 96-well cell culture plates were added. Each well of 4 wells was changed every 4 days. The same concentration of liquid, cultured for 12 days, observed the toxicity of drugs on the cells, set no drug cell control. 3 Drugs against HBsAg, HBeAg inhibition test: 2-fold diluted test solution below non-toxicity concentration, 3 dilutions of 4, 2, 1 and 0.5 g/L, 3 wells per concentration, and 37°C 50 mL/L CO 2 culture The culture medium was collected every 4 days. The culture medium was changed to the original concentration, and the HBsAg and HBeAg were measured simultaneously in the culture fluids collected on d8 and d12. 4HBsAg and HBeAg assay: Determined using a solid-phase radioimmunoassay kit. For instructions, see instructions. Determine the cpm value of each well using a gamma-counter. Results QL 4g/L was added to cell culture for 8 days and 12 days. The average half-toxicity (TC50) of three batches of experiments was 8.7g/L±0.3g/L, and the maximum non-toxic concentration (TC0) was 4g/L. The average inhibition rate of HBeAg secretion (8d) was 51.1%±2% and (12d) was 65.0%±1.9% at the maximum non-toxic concentration (4g/L), and the average effective dose (IC_(50)) was 8d. ) is 3.1g/L ± 0.8g/L and (12d) is 9.1g/L ± 0.9g/L, with