摘要
目的利用马丁达比犬的肾近曲小管上皮细胞(MD-CK)研究葛根素跨膜转运的机制。方法将MDCK按5×104个/cm2接种在悬挂式培养皿上培养至细胞生长分化成符合要求的细胞单层,用Millicell-ERS测量细胞单层的电阻值以确定细胞单层的完整性和致密性,改变葛根素的浓度、转运方向、温度以及加入P-糖蛋白(P-gp)抑制剂维拉帕米进行跨膜转运实验并用高效液相色谱(HPLC)测定葛根素的含量,计算表观渗透系数(Papp)分析葛根素的跨膜转运机制。结果 80 mg/L葛根素在37℃和4℃下,A→B方向上的Papp分别是(2.36±0.12)×10-6、(6.10±0.36)×10-7cm/s,而在B→A方向上则分别为(9.24±0.75)×10-7、(2.14±0.25)×10-7 cm/s。37℃下当加入维拉帕米之后80 mg/L葛根素的吸收Papp上升到(5.74±0.28)×10-6cm/s。结论葛根素的跨膜过程有多种转运机制参与,不仅包括简单扩散还有能量依赖的主动转运,同时呈现出明显的方向性,P-gp对葛根素的吸收有抑制作用。
Objective To study the transport were seeded 50 000 per cm： on Millicell-CM mechanism of puerarin based on MDCK. Methods MDCK cells Hangi values （TEER） were measured with Millicell ERS ng Culture Plates Inserts. Transepithelial electrical resistance （Electrical Resistance System） to indicate the integrity and compactness. The transport mechanism of puerarin was investigated by changing concentration, transport direction, temperature, and with or without verapamil. Puerarin was detected by a HPLC method and Papp was calculated. Re- suits Papp of 80 mg/L puerarin were （2. 36 ± 0. 12） x 10 -6 and （6. 10 ±0. 36） x 10 -7 cm/s in the basolateral compartment, 37C and 4C, while （9.24 ±0. 75） x 10-Tand （2. 14 ±0. 25） x 10 -7 cm/s in the apical compart- ment, respectively. Added with verapamil, P^vp of 80 mg/L raised to （5.74 ±0. 28） x 10-6 cm/s in the basolateral compartment. Conclusion The transport mechanism across MDCK includes not only simple diffusion but also ac- tive transport. The process shows obvious directionality. The efflux function of P-gp reduces absorption of puerarin.
出处
《安徽医科大学学报》
CAS
北大核心
2012年第12期1418-1422,共5页
Acta Universitatis Medicinalis Anhui
