利用Caco-2细胞单层模型预测柯楠碱、育亨宾、阿马碱、萝芙木碱的人肠吸收

Studies on predict of absorption of corynanthine,yohimbine,ajmalicine and ajmaline across human intestinal epithelial by using human Caco-2 cells monolayers

目的:研究中药化学成分柯楠碱(corynanthine,COR)、育亨宾(yohimbine,YOH)、阿马碱(ajmalicine,AMC)和萝芙木碱(ajmaline,AML)在人肠Caco-2细胞单层模型的转运吸收。方法:利用人源结肠腺癌细胞系Caco-2细胞单层模型研究COR,YOH,AMC和AML由绒毛面(AP侧)到基底面(BL侧),BL侧到AP侧两个方向的转运吸收过程。应用高效液相色谱法分离、紫外检测法检测对上述4个生物碱进行定量分析,计算转运参数和表观渗透系数(Papp),并与在Caco-2细胞单层模型上呈易吸收的阳性对照药普萘洛尔和难吸收的阳性对照药阿替洛尔的进行比较。用药物的ADMET预测软件,考察4个化合物的Papp值与其log D值之间的关系。结果:COR,YOH,AMC和AML由AP侧到BL侧的Papp值分别为(1.863±0.055)×10-5,(1.540±0.082)×10-5,(2.522±0.246)×10-5,(1.155±0.099)×10-5cm.s-1;由BL侧到AP侧的Papp值分别为(2.390±0.017)×10-5,(1.987±0.154)×10-5,(1.374±0.260)×10-5,(2.418±0.124)×10-5cm.s-1,与普萘洛尔的Papp A→B值[(2.23±0.10)×10-5cm.s-1]基本一致。COR,YOH,AMC和AML的Papp B→A/Papp A→B分别为1.28,1.29,0.54和2.09;AML外流是摄取的2.09倍。结论:COR,YOH,AMC和AML能够通过Caco-2细胞单层被动吸收,属于良好吸收的化合物。其中,AML在Caco-2细胞单层模型的转运可能存在外流机制。在4个生物碱的转运吸收过程中,油/水分配系数起着关键性的作用。

Objective： To predict the absorption of corynanthine （COR）, yohimbine （YOH）, ajmalicine （AMC） and ajmaline （AML） as chemical constituents of some traditional Chinese medicines in human intestinal epithelial. Method： By using Caco-2 （the human colonic adenocarcinoma cell lines） cell monolayers as a human intestinal epithelial cell model, the permeability of COR, YOH, AMC and AML were studied from apical side （ AP side） to basolateral side （ BL side） or from BL side to AP side. The four alkaloids were measured by high performance liquid chromatography （HPLC） coupled with UV detector. Transport parameters and apparent permeability coefficients （ Papp ） were calculated and compared with those of propranolol （ a control substance of high permeability） and atenolol （a control substance of poor permeability）. The relationship between Papp and log D values of four alkaloids was investigated by using drugs ADMET predict software. Result： The Papp values of COR, YOH, AMC and AML were （1. 863 ± 0. 055 ）×10^-5, （1.540±0.082）×10^-5, （2.522 ± 0.246） × 10^-5 and （ 1.155 ±0. 099） × 10^-5cm·s^-1 from AP side to BL side, and （2.390±0.017）×10^-5, （1.987±0.154）×10^-5, （1.374±0.260）×10^-5 and （2.418±0.124）×10^-5 cm·s^-1 from BL side to AP side, respectively, which Papp values were identical with that of propranolol [ （2. 23 ±0. 10） × 10^-5 cm·s^-1 from AP to BL side]. The ratio of Papp B→A/Papp A→B of COR, YOH, AMC and AML were 1.28, 1.29, 0. 54 and 2.09, respectively, which suggested that the efflux transport of AML was 2.09 times higher more than its influx transport. Condusion： COR, YOH, AMC and AML can be transported and absorbed across the human Caco-2 cells monolayers, and they belong to completely absorbed compounds. AML may have been involved in efflux mechanism in Caco-2 cells monolayers model from the BL to AP side direction. The oil-water partition coefficient play key roles in the transport and absorption of the four alkaloids.