醋柴胡多糖联用奥沙利铂增效活性及机制研究  

Synergistic activity and mechanism of vinegar baked radix bupleurum polysaccharides in combination with oxaliplatin

作　　者：韩明慧 王小双 赵亚 吴亚运 刘丽娟 赵瑞芝 Han Minghui;Wang Xiaoshuang;Zhao Ya;Wu Yayun;Liu Lijuan;Zhao Ruizhi(State Key Laboratory of Dampness Syndrome of Chinese Medicine,The Second Affiliated Hospital of Guangzhou University of Chinese Medicine;The Second Clinical Medical College of Guangzhou University of Chinese Medicine;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome)

机构地区：[1]广州中医药大学第二附属医院省部共建中医湿证重点实验室,广州510006 [2]广州中医药大学第二临床医学院,广州510006 [3]广东省中医证候临床研究重点实验室,广州510006

出　　处：《重庆医科大学学报》2025年第3期303-310,共8页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:82173981,82074086);广东省科技计划资助项目(编号:2023B1212060063,2021A1515110846)。

摘　　要：目的:探讨醋柴胡多糖联用奥沙利铂(oxaliplatin,OXA)增效活性及机制,为临床治疗原发肝细胞癌提供新的思路。方法:噻唑蓝(methyl thiazolyl tetrazolium,MTT)法检测醋制柴胡多糖3-4(vinegar baked radix bupleurum polysaccharides 3-4,VBCP 3-4)、醋制柴胡多糖3-3(vinegar baked radix bupleurum polysaccharides 3-3,VBCP 3-3)联用OXA对Huh7细胞的细胞杀伤作用;电感耦合等离子体质谱(inductively coupled plasma mass spectrometry,ICP-MS)检测Huh7细胞对OXA的摄取率,评价醋柴胡多糖VBCP 3-4联用OXA的体外增效途径;蛋白印迹法测定Huh7相关转运蛋白的表达,探讨VBCP 3-4联用OXA的增效机制。结果:VBCP 3-4、VBCP 3-3能够增加OXA对细胞的生长抑制作用。VBCP 3-4能够促进Huh7细胞对OXA的摄取,摄取率在4 h达到峰值,高、低剂量联用组增加摄取率为:分别43.43%(P=0.000)和29.02%(P=0.000)。OXA低、高剂量组单独作用Huh7细胞,能够促进细胞多药耐药相关蛋白(multidrug resistance-associated protein,MRP)2和MRP1蛋白表达,MRP2蛋白的上调幅度为18.11%、25.00%(P=0.008、P=0.001),MRP1蛋白的上调幅度为28.51%(P>0.05)、39.70%(P=0.015)。VBCP 3-4联用OXA后,MRP2、MRP1及乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)蛋白表达受到抑制,联用高、低剂量组MRP2降低幅度分别为47.38%、15.18%(P=0.000、P=0.049),MRP1蛋白降低幅度分别为64.96%、34.63%(P=0.000、P=0.000),联用高剂量组BCRP蛋白表达降低幅度为29.00%(P=0.020);VBCP 3-4单独作用Huh7细胞后,能够明显降低MRP2、MRP1及BCRP蛋白表达,VBCP 3-4高剂量组下调MRP2、MRP1幅度为24.91%、20.79%(P=0.004、P=0.005)。VBCP3-4下调BCRP蛋白的表达,高、中剂量下调幅度分别为15.02%、13.92%(P=0.003、P=0.030)。结论:醋柴胡多糖VBCP3-4通过抑制外排型转运蛋白MRP1、MRP2、BCRP表达,促进Huh7细胞摄入OXA,提高胞内有效浓度实现增效作用。Objective:To investigate the synergistic activity and mechanism of vinegar baked radix bupleurum polysaccharides(VBCP)in combination with oxaliplatin(OXA),and to provide new ideas for the clinical treatment of primary hepatocellular carcinoma.Methods:MTT assay was used to detect the cytotoxic effect of VBCP 3-4 and VBCP 3-3 in combination with OXA on Huh7 cells;ICP-MS was used to measure the uptake rate of OXA by Huh7 cells and evaluate the in vitro synergistic pathway of VBCP 3-4 in combination with OXA;Western blotting was used to measure the expression levels of related transporter proteins in Huh7 cells and explore the synergistic mechanism of VBCP 3-4 in combination with OXA.Results:VBCP 3-4 and VBCP 3-3 enhanced the inhibitory effect of OXA on the growth of cells.VBCP 3-4 promoted the uptake of OXA by Huh7 cells,with the peak uptake rate at 4 hours,and the uptake rate was increased to 43.43%in the high-dose combination group(P=0.000)and 29.02% in the low-dose combination group(P=0.000).Acting on Huh7 cells,the low-and high-dose OXA alone could promote the protein expression levels of MRP2 and MRP1 in Huh7 cells,and the protein expression level of multidrug resistance-associated protein(MRP)2 was upregulated to 18.11%and 25.00%,respectively(P=0.008,P=0.001),while that of MRP1 was upregulated to 28.51%(P>0.05)and 39.70%(P=0.015),respectively.After the combination of VBCP 3-4 and OXA,the protein expression of MRP2,MRP1,and breast cancer resistance protein(BCRP)was inhibited;MRP2 was reduced by 47.38%in the high-dose combination group(P=0.000)and 15.18%in the low-dose combination group(P=0.049);MRP1 was reduced by 64.96%in the high-dose combination group(P=0.000)and 34.63%in the low-dose combination group(P=0.000);BCRP was reduced by 29.00%(P=0.020)in the high-dose combination group.Acting on Huh7 cells alone,VBCP 3-4 significantly reduced the protein expression levels of MRP2,MRP1,and BCRP,and in the high-dose VBCP 3-4 group,MRP2 and MRP1 were reduced by 24.91%and 20.79%,respectively(P=0.004,P=0.005).VBCP 3-4

关 键 词：醋柴胡多糖 奥沙利铂 联用增效 抗肿瘤 

分 类 号：R285[医药卫生—中药学]