石菖蒲、安息香和苏合香挥发油促进罗丹明123的肠吸收作用及其机制

目的 研究石菖蒲、安息香和苏合香挥发油促进罗丹明-123（Rho-123）的肠吸收作用及其机制。方法 制作大鼠肠外翻模型，向肠囊内注入1.0 mL K-R营养液，将肠囊悬浮于45.0 mL 37℃的K-R溶液中。向肠囊外溶液中分别加入浓度均为60 μg/mL的石菖蒲、安息香和苏合香挥发油，孵育30 min后加入Rho-123，用高效液相色谱-荧光检测法测定肠囊内Rho-123的含量。培养人结肠癌细胞Caco-2，分别加入浓度均为80 μg/mL的石菖蒲、安息香和苏合香挥发油作用48 h，应用流式细胞术检测P糖蛋白（P-gp）的表达；采用qPCR检测P-gp基因MDR1 mRNA的表达。结果 石菖蒲、安息香和苏合香挥发油均可增加大鼠空肠和回肠Rho-123的吸收速率常数和表观通透系数（P<0.01），降低P-gp及其基因MDR1 mRNA的表达（3者作用的降低率分别为53.15%、55.10%、61.86%及55.41%、16.24%、38.46%；P<0.01）。结论 抑制P-gp及其基因的表达可能是石菖蒲、安息香和苏合香挥发油促进Rho-123肠吸收的主要机制之一。
Objective To explore the intestinal absorption-promoting effect of volatile oils of Shichangpu, benzoin and storax on Rhodamine-123 (Rho-123) and its mechanisms. Methods Rat in vitro everted gut sac (EGS) model was made. The EGS was filled with 1.0 mL K-R solution, and suspended in 45.0 mL 37℃ K-R solution with or without 60 μg/mL volatile oils of Shichangpu, benzoin or storax. After incubation for 30 min, Rho-123 was added in the experimental solution outside the EGS. Rho-123 content in the EGS was determined by high-performance liquid chromatography-fluorescence detector. The human colon carcinoma cell lines Caco-2 were cultured in DMEM and incubated with 80 μg/mL volatile oils of Shichangpu, benzoin or storax for 48 h. P-glycoprotein (P-gp) expression in Caco-2 cells was assessed using flow cytometry, and P-gp gene MDR1 mRNA expression was determined by qPCR. Results The volatile oils of Shichangpu, benzoin and storax each significantly increased the absorption rate constant and apparent permeability coefficient of Rho-123 in rat jejunum and ileum (P<0.01), reduced P-gp expression and MDR1 mRNA level in Caco-2 cells (the decreased rates of P-gp expression were 53.15%, 55.10% and 61.86%, and of MDR1 mRNA levels were 55.41%, 16.24% and 38.46%, P<0.01). Conclusion Inhibiting the protein and MDR1 mRNA expressions of P-gp may be one of the main mechanisms of volatile oils of Shichangpu, benzoin and storax in promoting Rho-123 intestinal absorption