多药耐药蛋白1a对苯甲酰新乌头原碱的效-毒-体内暴露的调控研究  

Study on the Regulation of Multidrug Resistance Protein 1a on the Efficacy-Toxicity-In Vitro Exposure of Benzoylmesaconine

作　　者：左慧琳 李小翠[1,2] 区晓君 杨彩华 刘中秋[1,2,4] 梁奇 朱丽君[1,2,4] ZUO Huilin;LI Xiaocui;OU Xiaojun;YANG Caihua;LIU Zhongqiu;LIANG Qi;ZHU Lijun(International Institute for Translational Chinese Medicine,School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Joint Laboratory for Translational Cancer Research of Chinese Medicine of the Ministry of Education of the People's Republic of China,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Department of Pharmacy,Nanfang Hospital,Southern Medical University,Guangzhou 510515 Guangdong,China;Guangdong-Hong Kong-Macao Joint Lab on Chinese Medicine and Immune Disease Research,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Shenzhen Bao'an Traditional Chinese Medicine Hospital,Shenzhen 518133 Guangdong,China)

机构地区：[1]广州中医药大学中药学院,国际中医药转化医学研究所,广东广州510006 [2]中华人民共和国教育部中医药防治肿瘤转化医学研究联合实验室,广东广州510006 [3]南方医科大学南方医院药学部,广东广州510515 [4]广州中医药大学粤港澳中医药与免疫疾病研究联合实验室,广东广州510006 [5]深圳市宝安区中医院,广东深圳518133

出　　处：《中药新药与临床药理》2023年第7期959-969,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家重点研发计划项目(2022YFC0867400);国家自然科学基金国际(地区)合作研究项目(81961128028);广东省自然科学基金杰出青年项目(2022B1515020079);广东省“岭南中医药”现代化基金项目(2020B1111100004);2020年广东省科技创新战略专项资金(粤港澳联合实验室)项目(2020B1212030006);深圳市“医疗卫生三名工程”项目(SZZYSM202206005)。

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^(-1)BMA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^(-1)BMA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^(-1)BMA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^(-1)BMA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�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^(-1)BMA,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^(-1)or gavage of 20 mg·kg^(-1)BMA 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^(-1)BMA,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 w

关 键 词：苯甲酰新乌头原碱 多药耐药蛋白 药效 毒性 体内暴露 小鼠 

分 类 号：R285.5[医药卫生—中药学]