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目的初步考察甘草提取物及其主要成分(甘草甜素、甘草次酸和甘草苷)对Caco-2细胞膜上P-糖蛋白(P-gp)功能和表达的影响,以备进一步探讨甘草新的解毒机制。方法利用流式细胞术检测Caco-2细胞内罗丹明123(Rho-123)的荧光强度和细胞膜上P-gp的表达,以考察P-gp外排功能和表达水平的变化。结果经过60min的干预,甘草提取物(1,10,100μg.mL-1)、甘草甜素(1,10,100μg.mL-1)和甘草苷(1,10,100μg.mL-1)Caco-2细胞内Rho-123的荧光强度较阴性对照组分别降低了34.66%、28.90%、24.56%、27.89%、26.29%、37.33%、19.51%、21.86%和19.03%(P<0.05)。经过72h的干预,甘草提取物中质量浓度(10μg.mL-1)组,甘草甜素低、中、高质量浓度(1,10,100μg.mL-1)组,甘草次酸中、高质量浓度(1,10μg.mL-1)组Caco-2细胞膜上P-gp表达的阳性率较阴性组分别增加了31.18%、61.67%、70.32%、77.43%、37.58%和49.14%(P<0.05)。结论甘草提取物、甘草甜素和甘草苷可能增强Caco-2细胞膜上P-gp的功能,而中质量浓度的甘草提取物,低、中、高质量浓度的甘草甜素和中、高质量浓度的甘草次酸则可能上调P-gp的表达。甘草甜素既能增强Ca-co-2细胞膜上P-gp的功能,又能上调其表达,可能是甘草影响P-gp的有效成分。
Objective To investigate the effect of licorice extract and its major components (glycyrrhizin, glycyrrhetinic acid and glycyrrhizin) on the function and expression of P-glycoprotein (P-gp) on Caco-2 cell membrane in order to further explore the new Detoxification mechanism. Methods Flow cytometry was used to detect the fluorescence intensity and the expression of P-gp on the membrane of Caco-2 cells in order to investigate the function and expression of P-gp. Results Caco-2 cells were treated with glycyrrhizin (1, 10, 100μg.mL-1), glycyrrhizin (1,10,100μg.mL-1) and glycyrrhizin (1,10,100μg.mL- Fluorescence intensity of Rho-123 decreased by 34.66%, 28.90%, 24.56%, 27.89%, 26.29%, 37.33%, 19.51%, 21.86% and 19.03%, respectively, compared with the negative control group (P <0.05). After 72h intervention, the concentration of glycyrrhizin in 10μg.mL-1 group, low, middle and high concentration of glycyrrhizin (1, 10, 100μg.mL-1) The positive rate of P-gp expression in Caco-2 cell membrane was increased by 31.18%, 61.67%, 70.32%, 77.43%, 37.58% and 49.14%, respectively (P <0.05) . Conclusion Licorice extract, glycyrrhizin and glycyrrhizin may enhance the function of P-gp on Caco-2 cell membrane. However, licorice extract at medium concentration, low, medium and high concentration of glycyrrhizin, medium and high concentration Glycyrrhetinic acid may up-regulate the expression of P-gp. Glycyrrhizin can enhance the function of P-gp on Ca-co-2 cell membrane and up-regulate the expression of P-gp, which may be the effective component of licorice on P-gp.