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目的:探讨连翘脂苷A对人克隆结肠癌细胞(Caco-2)细胞膜上P-糖蛋白(P-gp)的外排功能及作用机制。方法:采用刃天青法检测不同质量浓度连翘脂苷A对Caco-2的细胞毒性作用。以P-gp底物罗丹明123(Rh-123)为荧光探针,采用流式细胞术评价不同质量浓度连翘脂苷A对Rh-123细胞蓄积的影响,考察不同质量浓度连翘脂苷A对P-gp三磷酸腺苷(ATP)酶活性的影响。结果:连翘脂苷A在1~80 mg·L-1时,与阴性组比较,差异无统计学意义,细胞存活率>90%,对Caco-2细胞形态无影响。连翘脂苷A的荧光强度较空白组显著增加,其中80 mg·L-1连翘脂苷A荧光强度(223.43±5.6)%增加最多。连翘脂苷A不同质量浓度均有抑制P-gp ATP酶耗能的作用。结论:连翘脂苷A可通过抑制P-gp ATP酶产生抑制P-gp外排的作用。
Objective: To investigate the efflux function and mechanism of P-glycoprotein (P-gp) on the cell membrane of human colon carcinoma cell line Caco-2. Methods: Cytotoxicity of forsythin A with different mass concentration on Caco-2 was tested by resazurin method. The effect of different concentrations of forsythiaside A on the accumulation of Rh-123 cells was assessed by flow cytometry using P-gp substrate Rhodamine 123 (Rh-123) as a fluorescent probe. The effects of different concentrations of forsythiaside A on P-gp adenosine triphosphate (ATP) enzyme activity. Results: The results showed that there was no significant difference between the forsythia liposide A and the negative group when the concentration of glycoside A was 1 ~ 80 mg · L-1, the cell survival rate was over 90%, which had no effect on the morphology of Caco-2 cells. The fluorescence intensity of forsythin A increased significantly compared with that of blank group, and the fluorescence intensity of forsythin A 80 mg · L-1 (223.43 ± 5.6)% increased most. Forsythin A different concentrations of inhibitory P-gp ATPase energy consumption role. Conclusion: Forsythiaside A can inhibit the efflux of P-gp by inhibiting the production of P-gp ATPase.