参芪扶正注射液通过肿瘤相关巨噬细胞提高人乳腺癌MDA-MB-231细胞对顺铂的敏感性

目的:观察参芪扶正注射液通过免疫调节提高顺铂化疗的敏感性,并探讨其作用机制。方法:采用噻唑蓝(MTT)比色法测定参芪扶正注射液1,10,100 m L·L^-1对细胞的敏感性,流式细胞法检测细胞凋亡率,酶联免疫印迹法(Western blot)检测凋亡相关蛋白的表达,酶联免疫吸附实验(ELISA)检测细胞因子表达水平。结果:参芪扶正注射液能显著抑制人乳腺癌细胞MDA-MB-231与THP-1共培养细胞生长,参芪扶正注射液10,100 m L·L^-1时,其细胞存活率分别为71.8%,59.9%;参芪扶正注射液10 m L·L^-1与顺铂联合运用,提高了共培养细胞对顺铂的敏感性,顺铂的半数抑制浓度(IC50)由30μmol·L^-1降低至15μmol·L^-1;联合运用参芪扶正注射液10 m L·L^-1和顺铂,比15μmol·L^-1顺铂诱导的细胞凋亡率增加了15.5%(P<0.05);联合用药均能明显抑制B细胞淋巴瘤-2(Bcl-2),B细胞淋巴瘤-w(Bcl-w)和细胞外大淋巴瘤(Bcl-xl)的表达,增加Bcl-2相关X蛋白(Bax)和人BH3结构域凋亡诱导蛋白(Bid)的表达(P<0.05);参芪扶正注射液降低顺铂诱导的白细胞介素-10(IL^-10)和前列腺素E2(PGE2)的释放(P<0.05)。结论:参芪扶正注射液通过调节免疫细胞改善了乳腺癌细胞MDA-MB-231对顺铂的敏感性,起到协同抑制肿瘤细胞增殖的作用。该研究为益气扶正防治肿瘤提供实验依据,为中医药缓解肿瘤耐药研究提供实验参考。

Effect of Shenqi Fuzheng Injection in Improving Sensitivity of Human Breast Cancer MDA-MB-231 Cells to Cisplatin Through Tumor Associated Macrophages

Objective:To investigate the effect of Shenqi Fuzheng injection on the sensitivity of cisplatin through immunomodulation,in order to explore its mechanisms.Method:The cell survival was measured by thiazolyl blue tetrazolium bromide(MTT),the rate of apoptosis was detected by flow cytometry,the expressions of apoptosis-related proteins were detected by Western blot,and the expression level of cytokines was detected by enzyme linked immunosorbent assay(ELISA).Result:Shenqi Fuzheng injection 1,10,100 m L·L^-1 significantly inhibited the growth of co-cultured cells,and the cell survival rate was 71.8%and 59.9%at the concentration of 10,100 m L·L^-1 respectively.Shenqi Fuzheng injection 10 m L·L^-1 combined with cisplatin significantly increased the sensitivity of co-cultured cells to cisplatin.The half maximal inhibitory concentration(IC50)of cisplatin was reduced from 30 to 15μmol·L^-1.The rate of apoptosis induced by the combined treatment increased by 15.5%compared with that of cisplatin 15μmol·L^-1(P<0.05).The expressions of B-cell lymphoma-2(Bcl-2),B-cell lymphoma-w(Bcl-w)and B-cell lymphoma-extra large(Bcl-xl)were inhibited,and the expressions of Bcl-2-associated X protein(Bax)and protein induced by BH3 domain(Bid)apoptosis were increased(P<0.05).Shenqi Fuzheng injection reduced the release of interleukin-10(IL^-10)and prostaglandin E2(PGE2)induced by cisplatin(P<0.05).Conclusion:Shenqi Fuzheng injection improves the sensitivity of MDA-MB-231 to cisplatin by regulating immune cells,and plays a synergistic role in inhibiting the proliferation of breast cancer cells.This study provides experimental basis for the prevention and treatment of tumors with Yiqi Fuzheng method,and experimental reference for the study of traditional Chinese medicine in alleviating drug resistance of tumors.