薄荷醇促进丹参素肠道吸收的作用与机制研究  

Effect and Mechanism of Menthol on Intestinal Absorption of Salvianic Acid A

作　　者：武娟娟 王爱娟 恽昊 刘文彬 王晖 WU Juan-juan;WANG Ai-juan;YUN Hao;LIU Wen-bin;WANG Hui(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China;Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances,School of Life Sciences and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学中药学院,广州510006 [2]广东药科大学生命科学与生物制药学院,广东省生物活性药物研究重点实验室,广州510006

出　　处：《中国药学杂志》2022年第22期1888-1894,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助(81072591)。

摘　　要：目的考察薄荷醇促进肠道吸收的作用及其分子机制。方法体外建立大鼠外翻肠囊模型和Caco-2细胞单层模型考察不同浓度薄荷醇对丹参素经肠吸收的影响。分别采用2、8、16μmol·L^(-1)薄荷醇,25μmol·L^(-1)维拉帕米和100μmol·L^(-1)吡咯烷二硫代氨基甲酸钠(ammonium pyrrolidine dithiocarbamate,PDTC)处理Caco-2细胞2 h后,Western blot法检测P-糖蛋白(P-gp)蛋白、IκBα蛋白、p65胞浆蛋白、p65核蛋白表达变化,定量实时聚合酶链式反应(Q-PCR)法检测MDR1基因和IκBα基因表达变化。结果0.1%薄荷醇可以明显提高丹参素的药物吸收速率常数(P<0.05);2μmol·L^(-1)薄荷醇组和维拉帕米组AP-BL P_(app)显著高于对照组(P<0.05),16μmol·L^(-1)薄荷醇组AP-BL P_(app)低于对照组(P<0.01),各组间BL-AP差异无统计学意义;Caco-2细胞给药2 h后,与空白组比较,2、8μmol·L^(-1)薄荷醇、100μmol·L^(-1)PDTC、25μmol·L^(-1)维拉帕米对MDR1 mRNA和P-gp蛋白表达均有显著的抑制作用(P<0.05),2μmol·L^(-1)薄荷醇、100μmol·L^(-1)PDTC、25μmol·L^(-1)维拉帕米对IκBαmRNA和IκBα蛋白表达均有显著的促进作用(P<0.01),2、8μmol·L^(-1)薄荷醇组、100μmol·L^(-1)PDTC组和25μmol·L^(-1)维拉帕米组p65核蛋白水平均有降低作用(P<0.05),p65胞浆蛋白水平均有升高作用(P<0.05)。结论低浓度薄荷醇对丹参素肠道吸收有显著的促进作用,机制可能与其调控NF-kB信号通路,抑制P-gp蛋白表达有关。OBJECTIVE To investigate the effect of menthol on intestinal absorption and its molecular mechanism.METHODS The rat everted intestinal sac model and Caco-2 cell monolayer model were established in vitro to investigate the effects of different concentrations of menthol on the intestinal absorption of salvianic acid A.After treating Caco-2 cells with 2μmol·L^(-1),8,16μmol·L^(-1)menthol,25μmol·L^(-1)verapamil,and 100μmol·L^(-1)ammonium pyrrolidine dithiocarbamate(PDTC)for 2 hours,P-gp protein,IκBαprotein,p65 cytoplasmic protein and p65 nucleoprotein were detected by Western blot,Q-PCR method was used to detect the changes of MDR1 gene and IκBαgene expression.RESULTS The 0.1%menthol could significantly increase the drug absorption rate constant of salvianic acid A(P<0.05);2μmol·L^(-1)menthol group and verapamil group AP-BL P_(app)were significantly higher than control group(P<0.05),and the 16μmol·L^(-1)menthol group AP-BL P_(app)was lower than control group(P<0.01);AP-BL P_(app)of the 16μmol·L^(-1)menthol group was lower than that of the control group(P<0.01),there was no statistical difference in BL-AP among groups.After Caco-2 cells were administered for 2 hours,compared with the blank group,the 2,8μmol·L^(-1)menthol,100μmol·L^(-1)PDTC,and 25μmol·L^(-1)verapamil significantly decreased the expression of MDR1 mRNA and P-gp protein(P<0.05),the 2μmol·L^(-1)menthol,100μmol·L^(-1)PDTC,and 25μmol·L^(-1)verapamil significantly increased the expression of IκBαmRNA and IκBαprotein(P<0.01),the p65 nucleoprotein level of the 2μmol·L^(-1)menthol group,8μmol·L^(-1)menthol group,the PDTC group and the verapamil group were reduced(P<0.05),and the p65 cytoplasmic protein level was increased(P<0.05).CONCLUSION Low concentration of menthol can significantly promote the intestinal absorption of salvianic acid A,and its mechanism may be related to its regulation of NF-kB signaling pathway and inhibition of P-gp protein expression.

关 键 词：薄荷醇 肠道吸收促进剂 P-糖蛋白 CACO-2细胞 丹参素 

分 类 号：R965[医药卫生—药理学]