摘要
目的:考察醋柴胡多糖对拉米夫定抗乙型肝炎病毒(HBV)的增效作用,并初步探讨其作用机制。方法:将不同浓度的醋柴胡多糖、拉米夫定及二者联合作用于人肝癌细胞HepG2.2.15,同时设立对照组,孵育48 h,酶联免疫吸附试验试剂盒检测细胞上清液中乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)分泌量,荧光探针定量聚合酶链式反应(PCR)检测细胞HBV脱氧核糖核酸(DNA)表达量,金(正均)氏公式定量分析醋柴胡多糖的增效作用。采用高效液相色谱法测定细胞内拉米夫定含量;Western blot法测定有机阳离子转运蛋白(OCT)1、OCT2、P糖蛋白(P-gp)和多药耐药蛋白2(MRP2)的表达量。结果:与拉米夫定单用组相比,醋柴胡多糖增加拉米夫定对HBsAg分泌的抑制作用,表现为相加作用、对HBeAg作用表现为协同增强,Q值达6.55、对HBV-DNA抑制作用表现为相加。醋柴胡多糖低剂量组、醋柴胡多糖低中剂量组、醋柴胡多糖低高剂量组可显著促进拉米夫定的摄取;醋柴胡多糖高剂量联用组可显著降低P-gp的表达;醋柴胡多糖单用及联用组均可显著提高OCT1的表达。结论:醋柴胡多糖可通过增加拉米夫定的摄取发挥协同抗HBV作用,其作用机制可能P-gp、OCT1有关。
Objective:To study the synergistic effect of polysaccharide from vinegar-baked radix bupleuri(VBCP)with lamivudine(LAM)against hepatitis B virus(HBV)and investigate the mechanism of action.Methods:VBCP,LAM,and VBCP combined with LAM of different concentrations were added into HepG2.2.15 cells.At the same time,the control group was set up and incubated for 48 h.The secretion levels of hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in the supernatant of the cells were detected by enzyme-linked immunosorbent assay kit,and fluorescence probe quantitative polymerase chain reaction(PCR)was used to detect HBV-deoxyribonucleic acid(DNA)expression in cells.Quantitative analysis of the synergistic effect of polysaccharide from VBCP was conducted by Jin(Zhengjun)formula.The intracellular LAM content was determined by high-performance liquid chromatography.The expression levels of organic cation transporter(OCT)1,OCT2,P-glycoprotein(P-gp),and multidrug resistance protein 2(MRP2)were determined by Western blot.Results:Compared with mono-LAM treatment,polysaccharide from VBCP increased the inhibitory effect of LAM on HBsAg secretion,showing an additive effect,and the effect on HBeAg showed a synergistic enhancement.Q value was 6.55,and the inhibitory effect on HBV-DNA was additive.Low-dose,low and middle-dose,and low and high-dose groups of polysaccharide from VBCP could significantly promote the uptake of LAM.The expression of P-gp was significantly decreased in the combined group with a high dose of polysaccharide from VBCP,and groups with polysaccharide from VBCP alone and VBCP combined with LAM significantly increased the expression of OCT1.Conclusion:Polysaccharide from VBCP can play a synergistic anti-HBV effect by increasing the uptake of LAM,and its mechanism may be related to P-gp and OCT1.
作者
刘丽娟
赵亚
吴亚运
赵瑞芝
LIU Lijuan;ZHAO Ya;WU Yayun;ZHAO Ruizhi(The Second Clinical Medical College of Guangzhou University of Chinese Medicine Guangdong Provincial Hospital of Chinese Medicine,Guangdong Academy of Traditional Chinese Medicine,Guangzhou 510006,China;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome,Guangzhou 510120,China)
出处
《世界中医药》
CAS
2023年第8期1079-1084,共6页
World Chinese Medicine
基金
国家自然科学基金面上项目(82173981)
广东省科技计划项目(2017A020211013
2017B030314166)。
关键词
醋柴胡多糖
乙型肝炎病毒
乙型肝炎E抗原
乙肝病毒脱氧核糖核酸
协同增效
拉米夫定
有机阳离子转运蛋白1
P糖蛋白
多药耐药蛋白2
Polysaccharide from vinegar-baked radix bupleuri(VBCP)
Hepatitis B virus(HBV)
Hepatitis B e antigen(HBeAg)
HBV-DNA
Synergistic effect
lamivudine(LAM)
Organic cation transporter 1(OCT1)
P-glycoprotein(P-gp)
Multidrug resistance protein 2(MRP2)
