【摘要】 目的 探究多药耐药蛋白1a（Multidrug resistance protein 1a,Mdr1a）对苯甲酰新乌头原碱（Benzoylmesaconine,BMA）的镇痛和抗炎活性、神经和心脏毒性以及体内暴露的调控作用。方法 Mdr1a^-/-和野生型FVB小鼠分别灌胃20 mg·kg^-1BMA后，采用醋酸致疼痛扭体模型和角叉菜胶诱导急性炎症模型来考察Mdr1对BMA的镇痛和抗炎活性的调控作用，同时采用脑和心脏组织病理切片和ELISA法考察Mdr1对BMA神经和心脏毒性的调控作用。Mdr1a^-/-和野生型FVB小鼠静脉注射1 mg·kg^-1或灌胃20 mg·kg^-1BMA后，采用超高效液相-质谱（UHPLC-MS/MS）技术测定小鼠各组织和血浆中BMA的浓度，考察Mdr1a对BMA组织分布及药动学特征的影响。采用在体单向肠灌流模型考察Mdr1a对BMA肠道吸收的影响。结果 口服20 mg·kg^-1BMA后，Mdr1a^-/-小鼠的疼痛扭体次数较野生型FVB小鼠下降60.82%（P<0.05），足肿胀率无明显变化。与野生型FVB小鼠相比，Mdr1a^-/-小鼠口服BMA 8 h后海马DG区和CA区可见明显的锥体细胞核固缩，S100B钙结合蛋白水平增加了0.60倍，脑和心脏中BMA的含量分别增加了3.12和2.64倍（P<0.05），但心肌组织未见异常，肌酸激酶水平也无明显变化。组织分布结果显示，静注BMA 0.5和2 h后，与野生型FVB小鼠相比，Mdr1a^-/-小鼠的脑、心脏、肾脏、结肠和血浆中BMA的含量均显著上升（P<0.05）。药动学实验结果表明，与野生型FVB小鼠相比，BMA在Mdr1a^-/-小鼠上的生物利用度（F）增加了4.53倍，同时，口服20 mg·kg^-1BMA后，药时曲线下面积(AUC_0-t)和AUC_0-∞分别增加了1.46和2.63倍，清除率（Cl）和表现分布容积（V_d）分别降低了66.13%和78.85%（P<0.05）。肠灌流实验结果表明，与野生型FVB小鼠相比，BMA在Mdr1a^-/-小鼠十二指肠上的有效表观渗透系数，吸收率和吸收量分别显著增加了4.00、3.96和3.96倍（P<0.05）。结论 Mdr1a通过改变BMA的组织蓄积、体内暴露量和肠道吸收，参与调节BMA的镇痛作用和神经毒性，但BMA的抗炎作用和心脏毒性可能与Mdr1a的缺失无关。更多还原

【Abstract】 Objective To investigate the effects of multidrug resistance protein 1a （Mdr1a） on the analgesic and anti-inflammatory activities,neurotoxicity and cardiotoxicity and in vivo exposure of benzoylmesaconine （BMA）.Methods After Mdr1a^-/- and wild-type FVB mice were orally gavaged with 20 mg·kg^-1BMA,the effects of Mdr1a on the analgesic and anti-inflammatory activities of BMA were determined by using acetic acid-induced writhing and carrageenan-induced acute inflammation models,respectively.The effects of Mdr1a on the neurotoxicity and cardiotoxicity of BMA were investigated by pathological sections and ELISA method.The effects of Mdr1a on the tissue distributions and pharmacokinetics of BMA were investigated,the concentrations of BMA in tissues and plasma of the mice were measured concurrently by UHPLC-MS/MS after intravenous injection of 1 mg·kg^-1or gavage of 20 mg·kg^-1BMA in Mdr1a^-/- and wild-type FVB mice.Besides,the effect of Mdr1 on the intestinal absorption of BMA was conducted by using single-pass intestinal perfusion model.Results After oral administration of 20 mg·kg^-1BMA,the writhing times of Mdr1a^-/- mice decreased by 60.82%（P<0.05） when compared to those of wild-type FVB mice,but no significant differences were found on the paw edema rate.When compared to wildtype FVB mice,obvious pyramidal cell nuclear contraction in the dentate gyrus （DG） and cornu ammonis （CA）regions of the hippocampus were observed in Mdr1a^-/- mice after oral administration of BMA for 8 hours.Meanwhile,the level of S100B increased by 0.60 times,and the amounts of BMA in the brain and heart of Mdr1a^-/- mice increased by 3.12 and 2.64 folds,respectively （P<0.05）.No abnormality in myocardial tissues and no significant change of creatine kinase level were found between the two groups of mice.The results of tissue distributions showed that the contents of BMA in the brain,heart,kidney,colon and plasma of Mdr1a^-/- mice increased remarkably after 0.5 and 2 hours of intravenous injection of BMA while compared to those of wild-type FVB mice （P<0.05）.The results of pharmacokinetic study showed that the F on Mdr1a^-/- mice increased by 4.53 times,compared to that on wild-type FVB mice.In the meantime,after oral administration of 20 mg·kg^-1BMA,AUC_0-t and AUC_0-∞of BMA on Mdr1a^-/- mice increased by 1.46 and 2.63 times,respectively,and the Cl and V_d decreased by 66.13%and 78.85%,respectively （P<0.05）,while compared to those in wild-type FVB mice.The results of intestinal perfusion experiments showed that the effective apparent permeability coefficient,absorption rate and absorption amount of BMA in the duodenum of Mdr1a^-/- mice significantly increased by 4.00,3.96 and 3.96 times,respectively,compared to those of wild-type FVB mice （P<0.05）.Conclusion Mdr1a regulates the analgesic effect and neurotoxicity of BMA by altering its tissue accumulations,in vivo exposure and intestinal absorption.Mdrla may have no effect on the anti-inflammatory effect and cardiotoxicity of BMA.更多还原