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目的:探讨补肾疏肝方对人肺癌A549细胞及其相关干性因子的影响。方法:SD雄性大鼠随机分为6组,每组10只,分别为补肾疏肝方低、高剂量组(15,30 g·kg-1),顺铂组(8 g·kg-1),低剂量联合顺铂组(联合低剂量组),高剂量联合顺铂组(联合高剂量组),正常组,顺铂组ip给予相应药物,补肾疏肝方ig给予相应药物,正常组给予等体积的生理盐水,制备大鼠含药血清;分别配制10%含补肾疏肝方低、高剂量、顺铂、低剂量联合顺铂、高剂量联合顺铂,分别与肺癌A549细胞共同培养,CCK8法检测补肾疏肝方含药血清对A549增殖的影响;流式细胞术检测含药血清对A549细胞周期的影响;Annexin V/PI双染流式细胞术观察含药血清对A549细胞凋亡的影响;荧光定量PCR检测含药血清对相关m RNA表达的影响;流式细胞术检测含药血清对CD133蛋白表达的影响。结果:补肾疏肝方以时间、剂量依赖方式抑制A549细胞增殖,与顺铂联合有协同增效作用,24,48,72 h高剂量联合顺铂组的抑制率分别为45.39%,54.76%及59.94%;与正常组比较,补肾疏肝方、联合、顺铂含药血清可促进A549细胞凋亡,随着浓度增加,G2/M期细胞的百分比明显增加,明显下调CD133,SOX-2,OCT-4 m RNA的表达,明显降低CD133蛋白表达量,均具有明显统计学差异(P<0.05,P<0.01)。结论:补肾疏肝方可抑制A549细胞增殖,促进细胞凋亡,与顺铂联合可下调CD133,SOX-2,OCT-4等干性因子的表达。
Objective: To investigate the effects of Bushen Shugan Recipe on human lung cancer A549 cells and its related factors. Methods: SD male rats were randomly divided into 6 groups with 10 rats in each group. The rats were given low dose and high dose of Bushen Shugan Fang (15,30 g · kg -1) and cisplatin (8 g · kg -1) , Low-dose combined cisplatin group (combined with low-dose group), high-dose combined with cisplatin group (combined with high-dose group), normal group, cisplatin group corresponding drug given ip, Bushen Shugan prescription ig given corresponding drugs, Equal volume of normal saline to prepare rat drug-containing serum; respectively, formulated with Bushen Shugan Fang low, high dose, cisplatin, low dose combined with cisplatin, high-dose cisplatin, lung cancer A549 cells co- CCK8 assay was used to detect the effect of serum containing Bushen Shugan Fang on the proliferation of A549 cells. Flow cytometry was used to detect the effect of drug-containing serum on A549 cell cycle. Annexin V / PI double staining flow cytometry was used to observe the apoptosis of A549 cells The effect of drug-containing serum on the expression of m RNA was detected by real-time PCR. The effect of serum containing serum on the expression of CD133 protein was detected by flow cytometry. Results: Bushen Shugan decoction could inhibit the proliferation of A549 cells in a dose-dependent and time-dependent manner. Synergistic effect was observed when combined with cisplatin. The inhibitory rates were 45.39%, 54.76% and 24.48,72 h, respectively. 59.94%. Compared with the normal group, the serum of Bushen Shugan Fang combined with cisplatin could promote the apoptosis of A549 cells. With the increase of concentration, the percentage of cells in G2 / M phase increased obviously, and the expressions of CD133, SOX-2, The expression of OCT-4 m RNA significantly decreased the expression of CD133 protein, with statistical significance (P <0.05, P <0.01). Conclusion: Bushen Shugan decoction can inhibit the proliferation of A549 cells and promote cell apoptosis. Combined with cisplatin, it can down-regulate the expression of CD133, SOX-2, OCT-4 and other dry factors.