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目的:探索复方浙贝颗粒(CZBG)药物血清联合阿霉素(ADR)在体外对P388细胞的增殖抑制作用及机制。方法:1制备高、中、低剂量CZBG药物血清,以不同血清给药浓度作用于小鼠急性淋巴细胞白血病P388细胞株,MTT法观察CZBG药物血清对P388的抑制作用;2以不同剂量及浓度的CZBG药物血清联合ADR与单用ADR比较,分析联合用药后对P388细胞的增殖抑制影响;3流式细胞仪(FCM)定量检测实验各组P388细胞内阿霉素荧光值;Annexin-V/PI双染法检测P388细胞早期凋亡率。结果:1CZBG药物血清组及空白血清组在5%、10%及20%浓度时随着浓度增加对P388细胞株逐渐显示出抑增殖趋向,未发现两者间有统计学差异。CZBG药物血清高、中、低剂量在20%血清给药浓度时配伍不同浓度的ADR作用48h有增强ADR效应的趋势,剂量越高,增效作用越明显。其中CZBG高剂量联合ADR 0.04μg/mL及0.2μg/mL组与单用ADR组比较有显著性差异(t值分别为3.38,7.17,P<0.05);CZBG高、中、低剂量联合ADR用药后ADR IC50由0.46μg/mL分别降至0.05、0.30、0.30μg/mL;2经ADR作用后,不同剂量组的P388细胞内ADR荧光强度明显高于空白血清组。低剂量组最高,中,高剂量组次之。中剂量组早期凋亡率最高,为22.6%,高剂量组次之,为18.2%,低剂量组为16.4%,空白血清组为15.7%。结论:复方浙贝药物血清有增强阿霉素对P388细胞的增殖抑制趋向,可能与增加细胞内阿霉素含量,促进细胞凋亡有关。
Objective: To investigate the inhibitory effect and mechanism of CZBG drug serum combined with doxorubicin (ADR) on the proliferation of P388 cells in vitro. Methods: 1 CZBG serum was prepared at high, medium and low doses and treated with different concentrations of serum in mouse acute lymphoblastic leukemia cell line P388. The inhibitory effect of CZBG serum on P388 was observed by MTT assay. CZBG drug serum combined ADR with ADR alone was used to analyze the effect of combination therapy on the proliferation inhibition of P388 cells.3 Flow cytometry (FCM) was used to quantitatively detect the fluorescence intensity of doxorubicin in P388 cells; Annexin-V / PI double staining was used to detect the early apoptosis rate of P388 cells. Results: The concentration of CZBG drug serum group and blank serum group at 5%, 10% and 20% concentration gradually showed inhibition of proliferation on P388 cell line. There was no significant difference between the two groups. CZBG drug serum high, medium and low doses of 20% serum concentration and compatibility of different concentrations of ADR 48h have the potential to enhance the trend of ADR, the higher the dose, the more obvious synergistic effect. CZBG high dose combination of ADR 0.04μg / mL and 0.2μg / mL group and ADR group were significantly different (t values were 3.38,7.17, P <0.05); CZBG high, medium and low doses of ADR ADR of IC50 decreased from 0.46μg / mL to 0.05,0.30,0.30μg / mL, respectively. 2 After ADR, the fluorescence intensity of ADR in P388 cells of different dose groups was significantly higher than that of blank serum group. The highest dose, the second dose and the second dose were the lowest dose. The highest apoptosis rate was 22.6% in the middle dose group, 18.2% in the high dose group, 16.4% in the low dose group and 15.7% in the blank serum group. Conclusion: The compound prescription of Euphausia senticosus serum can enhance the inhibitory effect of doxorubicin on the proliferation of P388 cells, which may be related to the increase of doxorubicin content and the promotion of cell apoptosis.