灵芝多糖肽对Caco-2细胞上P糖蛋白功能、表达的影响

Effect of ganoderma lucidum polysaccharides peptide on the expression and function of p-glycoprotein in Caco-2 cells

目的研究灵芝多糖肽(ganoderma lucidum polysaccharides peptide,GLPP)对Caco-2细胞P糖蛋白(P-gp)功能和表达的影响。方法用流式细胞仪测定Caco-2细胞中P-gp底物罗丹明123和抗体的荧光强度,分析药物对P-gp功能和表达的影响。双向跨膜转运实验考查GLPP对罗丹明123转运的影响。结果 10、20、50、100μg.mL-1的GLPP均抑制了Rh-123的外排,其外排分别降低了8%、16%、19%、33%,具有浓度依赖性。高、中浓度的GLPP(50、100μg.mL-1)对Rh-123的双向转运有一定的抑制,外排率(ER)较阴性对照组降低了15%、18%。细胞与药物作用72h后,细胞抗体荧光强度降低,P-gp的表达降低。结论 GLPP对P-gp的活性有一定的抑制作用,能在较短的时间内增加细胞内Rh-123的蓄积,抑制Rh-123的双向转运。长期使用GLPP可抑制P-gp的表达。

Objective To determine the effect of ganoderma lucidum polysaccharides peptide （GLPP） on the expression and function of p-glycoprotein in Caco-2 cells. Methods The effect of GLPP on P-glycoprotein function and expression in Caeo-2 cells was analyzed by rhodanmine-123 and antibody assay. Flowcytometry was used to determine the intraeel- lular Rhodanmine-123 concentration and antibody concentration. The effect of GLPP on the transportation of rhodan- mine-123 was detected by Caco-2 cell monolayer model. Results Compared with that of the controls, the efflux quan- tity of rhodanmine-123 was reduced by 8~, 16~~, 19~//00 and 33~. when treated with GLPP at 10, 20, 50 and 100 /~g＂ mL-1~ The inhibitive effect of GLPP was concentration-dependent. Rhodanmine-123 bidirectional transport showed that the efflux rate of rhodanmine-123 was reduced by 15% and 18% when treated with GLPP at 50 and 100 yg ~ mL-1. The P-gp protein level was decreased after 72 h incubation. Conclusion GLPP is an inhibitor of P-gp which can inhibit the function of P-gp in 1 h and decrease the expression of P-gp after 72 h incubation with Caeo-2 cells. GLPP can also reduce the bidirectional transport of rhodanrnine-123.