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目的:研究芍药甘草汤(SGT)对Caco-2细胞P-糖蛋白(P-gp)的功能和表达的影响。方法:采用Caco-2单层细胞模型,以P-gp底物3H-地高辛为探针,P-gp抑制剂维拉帕米(Ver)为阳性对照药,测定SGT对Caco-2细胞P-gp功能的影响;用免疫组化法检测SGT对Caco-2细胞膜上的P-gp表达的影响。结果:在Caco-2模型上,3H-地高辛从基底侧(BL)到肠腔侧(AP)与肠腔侧(AP)到基底侧(BL)的表观渗透系数(apparent permeabilities,Papp)比值Papp(BL→AP)/Papp(AP→BL)约为27倍,证明3H-地高辛在Caco-2模型上的吸收方式为主动吸收。阳性药Ver与3H-地高辛联合时,明显增高了3H-地高辛在AP→BL方向的Papp(BL→AP)达3.82倍,但对BL→AP方向的转运影响不明显,说明Ver可明显抑制P-gp活性。SGT在1/25 IC5,1/5IC5,IC5浓度范围对P-gp底物3H-地高辛从AP→BL方向的转运分别促进了159.83%,217.95%,160.26%。1/25 IC5,1/5 IC5浓度下对3H-地高辛在BL→AP方向的转运分别促进了59.16%,50.73%,而相对于IC5浓度下SGT对3H-地高辛在BL→AP方向的转运则无影响。免疫组化结果显示SGT对P-gp的表达有抑制作用。结论:SGT可抑制P-gp的功能和表达,从而促进P-gp底物的吸收。
Objective: To investigate the effect of Shaoyao Gancao Decoction (SGT) on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. Methods: Caco-2 monolayer cell model was used to detect the inhibitory effect of SGT on Caco-2 cells by using P-gp substrate 3H-digoxin as probe and P-gp inhibitor Verapamil as positive control. The effect of SGT on the expression of P-gp on Caco-2 cell membrane was detected by immunohistochemistry. RESULTS: In the Caco-2 model, the apparent permeabilities (Papp) of 3H-digoxin from the basolateral (BL) to the luminal (AP) and basal (BL) ) Ratio Papp (BL → AP) / Papp (AP → BL) was about 27 times, which proves that the absorption mode of 3H-digoxin in the Caco-2 model is active absorption. When the positive drug Ver combined with 3H-digoxin, 3H-digoxin significantly increased Papp (BL → AP) by 3.82 times in the AP → BL direction but had no obvious effect on the transport of BL → AP, indicating that Ver P-gp activity can be significantly inhibited. SGT promoted the transport of P-gp substrate 3H-digoxin from AP → BL by 159.83%, 217.95% and 160.26% in the range of 1/25 IC5, 1/5 IC5 and IC5, respectively. At 1/5 IC5 and 1/5 IC5 concentrations, the translocation of 3H-digoxin in the BL → AP direction was promoted by 59.16% and 50.73%, respectively. However, compared with the SGT at IC5 concentration, 3H- The direction of transport has no effect. Immunohistochemistry showed that SGT inhibited the expression of P-gp. Conclusion: SGT can inhibit the function and expression of P-gp and promote the absorption of P-gp substrate.