通脉方中异黄酮类化合物在人源肠Caco2细胞模型的吸收转运(1)
[摘要]通脈方是由葛根、丹参和川芎3味药按质量1∶1∶1组成的复方。该文研究通脉方中异黄酮类化合物大豆苷元、芒柄花素、5羟基芒柄花苷、芒柄花苷、大豆苷、3′甲氧基葛根素、染料木苷、葛根素、芒柄花素8CβD呋喃芹糖基(1→6)OβD吡喃葡萄糖苷、芒柄花素7OβD呋喃芹糖基(1→6)OβD吡喃葡萄糖苷、澳白檀苷、葛花宁、大豆苷元7，4′二OβD吡喃葡萄糖苷、泰国野葛根素、3′羟基葛根素、3′甲氧基大豆苷、芒柄花素8CβD吡喃木糖基(1→6)OβD吡喃葡萄糖苷、染料木素8CβD呋喃芹糖基(1→6)OβD吡喃葡萄糖苷、染料木素7OβD呋喃芹糖基(1→6)OβD吡喃葡萄糖苷、3′羟基泰国野葛根素、6″OβD木糖基葛根素、鹰嘴豆芽素A8CβD呋喃芹糖基(1→6)OβD吡喃葡萄糖苷、3′甲氧基大豆苷元7，4′二OβD吡喃葡萄糖苷、大豆苷元7OβD吡喃葡萄糖基(1→4)OβD吡喃葡萄糖苷和大豆苷元7OαD吡喃葡萄糖基(1→4)OβD吡喃葡萄糖苷在肠的吸收转运。采用人源肠Caco2细胞单层模型，研究通脉方中上述25个异黄酮类化合物由绒毛面(AP侧)到基底面(BL侧)或从BL侧到AP侧2个方向的转运过程。应用高效液相色谱法分离、紫外检测法对化合物进行定量分析，计算表观渗透系数(Papp)，并与阳性对照药普萘洛尔和阿替洛尔比较。大豆苷元和芒柄花素由AP侧到BL侧的Papp分别为(255±003)×10-5，(306±001)×10-5 cm·s-1；由BL侧到AP侧的Papp分别为(262±0)×10-5，(265±011)×10-5 cm·s-1。与本试验中在Caco2细胞单层模型上呈良好吸收的阳性对照药普萘洛尔的Papp(266±032)×10-5 cm·s-1和呈难吸收的阳性对照药阿替洛尔的Papp(234±010)×10-7 cm·s-1比较，大豆苷元和芒柄花素与普萘洛尔在同一数量级；其他化合物与阿替洛尔在同一数量级。大豆苷元和芒柄花素的Papp AP→BL/Papp BL→AP分别为097，115。可以预测，大豆苷元和芒柄花素可以通过小肠上皮细胞被动吸收进入体内，属于良好吸收的化合物；其他化合物属于吸收不良的化合物。5羟基芒柄花苷、染料木苷、澳白檀苷、葛花宁和染料木素7OβD呋喃芹糖基(1→6)OβD吡喃葡萄糖苷的Papp AP→BL/Papp BL→AP分别为018，028，045，038，049，推测它们在Caco2细胞单层模型中的转运可能存在外流机制。
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[关键词]Caco2细胞单层模型；通脉方；异黄酮；大豆苷元；芒柄花素；肠吸收；表观渗透系数
Absorption and transport of isoflavonoid compounds from Tongmai
formula across human intestinal epithelial (Caco2) cells in vitro
WANG Furong， YANG Xiuwei*
(State Key Laboratory of Natural and Biomimetic Drugs， Department of Natural Medicines，School of Pharmaceutical Sciences， Peking University， Beijing 100191， China)
, 百拇医药
[Abstract]Tongmai formula (TMF) is a drug combination of three components including Puerariae Lobatae Radix [roots of Pueraria lobata]， Salviae Miltiorrhizae Radix (roots of Salvia miltiorrhiza) and Chuanxiong Rhizoma (rhizomes of Ligusticum chuanxiong) in a weight ratio of 1∶1∶1 The absorption and transport of isoflavonoid compounds from Tongmai formula across human intestinal epithelial (Caco2) cells in vitro were studied in this paper The assay isoflavonoid compounds include daidzein， formononetin， 5hydroxylononin， ononin， daidzin， 3′methoxypuerarin， genistin， puerarin， formononetin8CβDapiofuranosyl(1→6)OβDglucopyranoside， formononetin7OβDapiofuranosyl(1→6)OβDglucopyranoside， lanceolarin， kakkanin， daidzein7，4′diOβDglucopyranoside， mirificin， 3′hydroxypuerarin， 3′methoxydaidzin， formononetin8CβDxylopyranosyl(1→6)OβDglucopyranoside， genistein8CβDapiofuranosyl(1→6)OβDglucopyranoside， genistein7OβDapiofuranosyl(1→6)OβDglucopyranoside (ambocin)， 3′hydroxymirificin， 6″OβDxylosylpuerarin， biochanin A8CβDapiofuranosyl(1→6)OβDglucopyranoside， 3′methoxydaidzein7，4′diOβDglucopyranoside， daidzein7OβDglucopyranosyl(1→4)OβDglucopyranoside， and daidzein7OαDglucopyranosyl(1→4)OβDglucopyranoside By using human Caco2 monolayer as an intestinal epithelial cell model in vitro， the permeability of abovementioned 25 isoflavonoids in TMF were studied from the apical (AP) side to basolateral (BL) side or from the BL side to AP side The assay compounds were determined by reversed phased highperformance liquid chromatography (HPLC) coupled with UV detector Transport parameters and apparent permeability coefficients (Papp) were then calculated and and compared with those of propranolol and atenolol， which are the transcellular transport marker and as a control substance for high and poor permeability， respectively The Papp values of daidzein and formononetin were (255±003) ×10-5，(306±001) ×10-5 cm·s-1 from AP side to BL side， respectively， and (262±000) ×10-5， (265±011) ×10-5 cm·s-1 from BL side to AP side， respectively Under the condition of this experiment， the Papp value was (266±032) ×10-5 cm·s-1 for propranolol and (234±010) ×10-7 cm·s-1 for atenolol The Papp values of daidzein and formononetin were at a same magnitude with those of propranolol And the Papp values of other 23 isoflavonoid compounds were at a same magnitude with those of atenolol On the other hand， the rats of Papp AP→BL/Papp BL→AP of daidzein and formononetin on the influx transport were 097 and 115， respectively It can be predicted that daidzein and formononetin can be absorbed across intestinal epithelial cells to go to the body circulation by the passive diffusion mechanism and they were assigned to the wellabsorbed compounds Other 23 isoflavonoid compounds were assigned to the poorly absorbed compounds Because of the rats of Papp AP→BL/Papp BL→AP of 5hydroxylononin， genistin， lanceolarin， kakkanin， and genistein7OβDapiofuranosyl(1→6)OβDglucopyranoside were 018， 028， 045， 038， 049， they may have been involved in the efflux mechanism in Caco2 cells monolayer model from the BL side to AP side direction, http://www.100md.com(王付荣 杨秀伟)
, http://www.100md.com
[关键词]Caco2细胞单层模型；通脉方；异黄酮；大豆苷元；芒柄花素；肠吸收；表观渗透系数
Absorption and transport of isoflavonoid compounds from Tongmai
formula across human intestinal epithelial (Caco2) cells in vitro
WANG Furong， YANG Xiuwei*
(State Key Laboratory of Natural and Biomimetic Drugs， Department of Natural Medicines，School of Pharmaceutical Sciences， Peking University， Beijing 100191， China)
, 百拇医药
[Abstract]Tongmai formula (TMF) is a drug combination of three components including Puerariae Lobatae Radix [roots of Pueraria lobata]， Salviae Miltiorrhizae Radix (roots of Salvia miltiorrhiza) and Chuanxiong Rhizoma (rhizomes of Ligusticum chuanxiong) in a weight ratio of 1∶1∶1 The absorption and transport of isoflavonoid compounds from Tongmai formula across human intestinal epithelial (Caco2) cells in vitro were studied in this paper The assay isoflavonoid compounds include daidzein， formononetin， 5hydroxylononin， ononin， daidzin， 3′methoxypuerarin， genistin， puerarin， formononetin8CβDapiofuranosyl(1→6)OβDglucopyranoside， formononetin7OβDapiofuranosyl(1→6)OβDglucopyranoside， lanceolarin， kakkanin， daidzein7，4′diOβDglucopyranoside， mirificin， 3′hydroxypuerarin， 3′methoxydaidzin， formononetin8CβDxylopyranosyl(1→6)OβDglucopyranoside， genistein8CβDapiofuranosyl(1→6)OβDglucopyranoside， genistein7OβDapiofuranosyl(1→6)OβDglucopyranoside (ambocin)， 3′hydroxymirificin， 6″OβDxylosylpuerarin， biochanin A8CβDapiofuranosyl(1→6)OβDglucopyranoside， 3′methoxydaidzein7，4′diOβDglucopyranoside， daidzein7OβDglucopyranosyl(1→4)OβDglucopyranoside， and daidzein7OαDglucopyranosyl(1→4)OβDglucopyranoside By using human Caco2 monolayer as an intestinal epithelial cell model in vitro， the permeability of abovementioned 25 isoflavonoids in TMF were studied from the apical (AP) side to basolateral (BL) side or from the BL side to AP side The assay compounds were determined by reversed phased highperformance liquid chromatography (HPLC) coupled with UV detector Transport parameters and apparent permeability coefficients (Papp) were then calculated and and compared with those of propranolol and atenolol， which are the transcellular transport marker and as a control substance for high and poor permeability， respectively The Papp values of daidzein and formononetin were (255±003) ×10-5，(306±001) ×10-5 cm·s-1 from AP side to BL side， respectively， and (262±000) ×10-5， (265±011) ×10-5 cm·s-1 from BL side to AP side， respectively Under the condition of this experiment， the Papp value was (266±032) ×10-5 cm·s-1 for propranolol and (234±010) ×10-7 cm·s-1 for atenolol The Papp values of daidzein and formononetin were at a same magnitude with those of propranolol And the Papp values of other 23 isoflavonoid compounds were at a same magnitude with those of atenolol On the other hand， the rats of Papp AP→BL/Papp BL→AP of daidzein and formononetin on the influx transport were 097 and 115， respectively It can be predicted that daidzein and formononetin can be absorbed across intestinal epithelial cells to go to the body circulation by the passive diffusion mechanism and they were assigned to the wellabsorbed compounds Other 23 isoflavonoid compounds were assigned to the poorly absorbed compounds Because of the rats of Papp AP→BL/Papp BL→AP of 5hydroxylononin， genistin， lanceolarin， kakkanin， and genistein7OβDapiofuranosyl(1→6)OβDglucopyranoside were 018， 028， 045， 038， 049， they may have been involved in the efflux mechanism in Caco2 cells monolayer model from the BL side to AP side direction, http://www.100md.com(王付荣 杨秀伟)