清解扶正颗粒促进凋亡和自噬逆转结直肠癌细胞多药耐药研究  

Qingjie Fuzheng Granules Reverse Multidrug Resistance of Colorectal Cancer Resistant Cells via Inducing Apoptosis and Autophagy

作　　者：赵锦燕 朱晓勤 蓝紫林 李斐 林明和 林久茂 ZHAO Jinyan;ZHU Xiaoqin;LAN Zilin;LI Fei;LIN Minghe;LIN Jiumao(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350012,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350012,China;Key Laboratory of Integrative Medicine of Fujian Province University,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中西医结合研究院,福州350012 [2]福建省中西医结合老年性疾病重点实验室,福州350012 [3]福建省中西医结合高校重点实验室,福州350122

出　　处：《世界中医药》2024年第15期2246-2252,共7页World Chinese Medicine

基　　金：国家自然科学基金项目(81303125);福建省自然科学基金项目(2021J01939,2022J01368)。

摘　　要：目的:探讨清解扶正颗粒(QFG)对人结直肠癌HCT-8/5-FU耐药细胞的逆转作用机制。方法:培养人结直肠癌HCT-8细胞及HCT-8/5-FU耐药细胞,计算不同化疗药物的耐药指数(RI)和QFG的逆转倍数(RF);设立空白对照组(0 mg/mL)和不同浓度(0.25、0.5、1.0 mg/mL)QFG干预组;各组干预48 h后,通过罗丹明-123(Rho123)染色和多柔比星(DOX)染色检测细胞外排泵功能,通过克隆形成试验检测细胞存活能力,应用磷脂酰丝氨酸(Annexin V-FITC)/碘化丙啶(PI)双染检测细胞凋亡,应用Cyto-ID染色检测细胞自噬,应用蛋白质印迹法检测P-糖蛋白(gP)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、选择性自噬接头蛋白(p62/SQSTM1)、微管相关蛋白1轻链(LC)3Ⅱ的蛋白表达。结果:HCT-8/5-FU细胞对5-氟尿嘧啶(5-FU)、多柔比星(DOX)和顺铂(DDP)均耐药,QFG可有效逆转化疗耐药,与空白对照组比较,不同浓度QFG显著增加Rho123、DOX和Cyto-ID的平均荧光强度(均P<0.05);抑制细胞的克隆形成率(P<0.05),提高细胞凋亡率(P<0.05);和空白对照组比较,QFG抑制P-gP、p62的蛋白表达(P<0.05),降低Bcl-2/Bax比值(P<0.05),促进LC3-Ⅱ的蛋白表达(P<0.05)。结论:QFG促进细胞凋亡和自噬,抑制耐药细胞对化疗药物的外排泵功能,从而逆转多药耐药。Objective:To investigate the mechanism by which Qingjie Fuzheng Granules(QFG)reverse drug resistance in human colorectal cancer HCT-8/5-FU resistant cells.Methods:HCT-8 cells and HCT-8/5-FU resistant cells were cultured,and the resistance index(RI)for different chemotherapeutic drugs and the reversal fold(RF)of QFG were calculated.The cells were divided into a blank control group(0 mg/mL)and QFG intervention groups with different concentrations(0.25,0.5,1.0 mg/mL).After 48 hours of intervention,the efflux pump function was assessed by rhodamine-123(Rho123)and doxorubicin(DOX)staining.Cell survival was evaluated by colony formation assay.Apoptosis was detected using Annexin V-FITC/propidium iodide(PI)double staining.Autophagy was analyzed using Cyto-ID staining.Protein expression levels of P-glycoprotein(P-gP),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),selective autophagy receptor protein(p62/SQSTM1),and microtubule-associated protein 1 light chain 3Ⅱ(LC3-Ⅱ)were measured by Western blot.Results:HCT-8/5-FU cells showed resistance to 5-fluorouracil(5-FU),DOX,and cisplatin(DDP).QFG effectively reversed this drug resistance.Compared with the blank control group,QFG significantly increased the average fluorescence intensity of Rho123,DOX,and Cyto-ID(all P<0.05),inhibited cell colony formation(P<0.05),and increased the apoptosis rate(P<0.05).QFG also reduced the protein expression of P-gP and p62(P<0.05),decreased the Bcl-2/Bax ratio(P<0.05),and promoted the protein expression of LC3-Ⅱ(P<0.05).Conclusion:QFG can promote apoptosis and autophagy,inhibit the efflux pump function of resistant cells,and thereby reverse multidrug resistance.

关 键 词：清解扶正颗粒 结直肠癌 多药耐药 细胞凋亡 细胞自噬 克隆形成 逆转 泵 

分 类 号：R242[医药卫生—中医临床基础] R574.6[医药卫生—中医学]