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目的:观察益气复元汤是否增加化疗药诱导大肠癌Lovo细胞凋亡的影响以及其分子水平作用机制。方法:试验分为益气复原汤、化疗药、益气复原汤+化疗药各自含药血清、血清对照组、空白对照组等5组,人吖啶橙/溴乙啶(AO/EB)双荧光染色法、Annexin V-FITC(FITC,异硫氰酸荧光素)单染法检测各组对大肠癌Lovo细胞凋亡诱导作用,Western blot(蛋白免疫印迹)法检测Lovo细胞凋亡蛋白B细胞淋巴瘤/白血病-2(Bcl-2),多肽抗原(Bax)的表达。结果:细胞形态学及流式细胞检测都表明益气复原汤含药血清对大肠癌Lovo细胞有诱导凋亡作用,并增强化疗药诱导癌细胞凋亡效应;Western blot法检测显示①血清对照组与空白对照组中Bcl-2,Bax蛋白表达无明显差异;②益气复元汤联合化疗组、化疗组、益气复元汤组、血清对照组各组Bcl-2,Bax蛋白表达组间两两比较具有显著差异,P<0.01;益气复元汤联合化疗组Bcl-2表达最弱,其次是化疗组,再次是益气复元汤组,血清对照组最强。③益气复元汤联合化疗组Bax表达最强,其次是化疗组,再次是益气复元汤组,血清对照组最弱。结论:益气复元汤对Lovo肠癌细胞有诱导细胞凋亡作用,并增强化疗药诱导癌细胞凋亡效应;下调Bcl-2蛋白及上调Bax蛋白的表达是诱导Lovo肠癌细胞凋亡的主要机制之一。
Objective: To observe whether Yiqi Fuyuan Decoction can increase the apoptosis induced by chemotherapeutics in colorectal cancer Lovo cells and its molecular mechanism. Methods: The experimental groups were divided into 5 groups: acridine orange / ethidium bromide (AO / EB) bis (acridine orange / ethidium bromide), chemotherapy drug, Yiqi Fuyuan decoction + chemotherapeutic drug containing serum, serum control group and blank control group The apoptosis of Lovo cells was detected by FITC and FITC staining. The apoptosis of B cells was detected by Western blot (Western blot) Lymphoma / leukemia-2 (Bcl-2), polypeptide antigen (Bax) expression. Results: The results of cell morphology and flow cytometry showed that Yiqi Fuyuan Decoction-containing serum could induce apoptosis of Lovo cells and enhance the apoptosis of cancer cells induced by chemotherapeutic drugs. The results of Western blot showed that ① serum control group There was no significant difference between Bcl-2 and Bax protein expression in the control group, Bcl-2 and Bax protein expression in the control group, Yiqi Fuyuan Decoction combined with chemotherapy group, chemotherapy group, Yiqi Fuyuan Decoction group and serum control group P <0.01. The expression of Bcl-2 in Yiqi Fuyuan Decoction combined with chemotherapy group was the weakest, followed by chemotherapy group and Yiqi Fuyuan Decoction group again. The serum control group was the strongest. ③ Yiqi Fuyuan Decoction combined with chemotherapy group Bax the strongest expression, followed by chemotherapy group, Yiqi Fuyuan Decoction group again, the serum control group weakest. Conclusion: Yiqi Fuyuan Decoction can induce cell apoptosis in Lovo cells and enhance the apoptosis of cancer cells induced by chemotherapeutic drugs. Down-regulation of Bcl-2 protein and up-regulation of Bax protein are the main inducers of apoptosis in Lovo cells One of the mechanisms.