Berberine reverses multidrug resistance in Candida albicans by hijacking the drug efflux pump Mdr1p  

小檗碱通过挟持药物外排泵Mdr1p逆转白色念珠菌的多药耐药

作　　者：Yaojun Tong Jingyu Zhang Nuo Sun Xiang-Ming Wang Qi Wei Yu Zhang Ren Huang Yingying Pu Huanqin Dai Biao Ren Gang Pei Fuhang Song Guoliang Zhu Xinye Wang Xuekui Xia Xiangyin Chen Lan Jiang Shenlin Wang Liming Ouyang Ning Xie Buchang Zhang Yuanying Jiang Xueting Liu Richard Calderone Fan Bai Lixin Zhang Gil Alterovitzk 童垚俊;张敬宇;孙诺;王湘铭;危期;张钰;黄韧;普颖颖;代焕琴;任彪;裴刚;宋福行;朱国良;王馨叶;夏雪奎;陈相因;蒋岚;王申林;欧阳立明;谢宁;张部昌;姜远英;刘雪婷;Richard Calderone;白凡;张立新;Gil Alterovitzk(State Key Laboratory of Bioreactor Engineering and School of Biotechnology,East China University of Science and Technology,Shanghai 200237,China;Chinese Academy of Sciences Key Laboratory of Pathogenic Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100190,China;Biomedical Pioneering Innovation Center(BIOPIC),School of Life Sciences,Peking University,Beijing 100871,China;Georgetown University Medical Center,Department of Microbiology&Immunology.Washington DC 20057,USA;Institute of Health Sciences,School of Life Sciences,Anhui University,Hefei 230601,China;Guangdong Provincial Key Laboratory of Laboratory Animals,Guangdong Laboratory Animals Monitoring Institute,Guangzhou 510663,China;Department of Pharmacology,Second Military Medical University,Shanghai 200433,China;State Key Laboratory of Mycology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;The Novo Nordisk Foundation Center for Biosustainability,Technical University of Denmark,2800 Kgs.Lyngby,Denmark;Key Biosensor Laboratory of Shandong Province,Biology Institute,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250013,China;Brigham and Women's Hospital,Boston MA 02115,USA;National Artificial Intelligence Institute,US.Department of Veterans Affairs,Washington DC 20420,USA)

机构地区：[1]State Key Laboratory of Bioreactor Engineering and School of Biotechnology,East China University of Science and Technology,Shanghai 200237,China [2]Chinese Academy of Sciences Key Laboratory of Pathogenic Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100190,China [3]Biomedical Pioneering Innovation Center(BIOPIC),School of Life Sciences,Peking University,Beijing J00871,China [4]Georgetown University Medical Center,Department of Microbiology&Immunology.Washington DC 20057,USA [5]Institute of Health Sciences,School of Life Sciences,Anhui University,Hefei 230601,China [6]Guangdong Provincial Key Laboratory of Laboratory Animals,Guangdong Laboratory Animals Monitoring Institute,Guangzhou 510663,China [7]Department of Pharmacology,Second Military Medical University,Shanghai 200433,China [8]State Key Laboratory of Mycology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China [9]The Novo Nordisk Foundation Center for Biosustainability,Technical University of Denmark,2800 Kgs.Lyngby,Denmark [10]Key Biosensor Laboratory of Shandong Province,Biology Institute,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250013,China [11]Brigham and Women's Hospital,Boston MA 02115,USA [12]National Artificial Intelligence Institute,US.Department of Veterans Affairs,Washington DC 20420,USA

出　　处：《Science Bulletin》2021年第18期1895-1905,M0004,共12页科学通报（英文版）

基　　金：supported by the National Key Research and Development Program of China (2020YFA0907800);the National Natural Science Foundation of China (31720103901);the “111” Project of China (B18022);the Fundamental Research Funds for the Central Universities (22221818014S);the Open Project Funding of the State Key Laboratory of Bioreactor Engineering,the Shandong Taishan Scholar Award,and the Novo Nordisk Foundation (NNF10CC1016517)。

摘　　要：Clinical use of antimicrobials faces great challenges from the emergence of multidrug-resistant pathogens. The overexpression of drug efflux pumps is one of the major contributors to multidrug resistance(MDR). Reversing the function of drug efflux pumps is a promising approach to overcome MDR. In the life-threatening fungal pathogen Candida albicans, the major facilitator superfamily(MFS) transporter Mdr1p can excrete many structurally unrelated antifungals, leading to MDR. Here we report a counterintuitive case of reversing MDR in C. albicans by using a natural product berberine to hijack the overexpressed Mdr1p for its own importation. Moreover, we illustrate that the imported berberine accumulates in mitochondria and compromises the mitochondrial function by impairing mitochondrial membrane potential and mitochondrial Complex I. This results in the selective elimination of Mdr1 p overexpressed C. albicans cells. Furthermore, we show that berberine treatment can prolong the mean survival time of mice with blood-borne dissemination of Mdr1p overexpressed multidrug-resistant candidiasis. This study provides a potential direction of novel anti-MDR drug discovery by screening for multidrug efflux pump converters.药物外排泵的过量表达是病原微生物产生多药耐药(MDR)的重要原因之一.逆转药物外排泵的功能有望对抗多药耐药问题.在能危及人类生命的真菌病原菌白色念珠菌中,主要易化子超家族(MFS)类型转运蛋白Mdr1p能非特异性地向细胞外转运很多结构上不相关的抗真菌化合物,从而导致了白色念珠菌的多药耐药.本研究报道了一个有悖过往认知的案例:一个天然产物小檗碱不仅不会被白色念珠菌Mdr1p外排,而且还能特异性地挟持过表达的Mdr1p把自己运输进细胞.进一步研究发现,进入白色念珠菌细胞后的小檗碱能够在线粒体中积累,通过破坏线粒体膜电势和线粒体复合物I引起线粒体功能紊乱,从而杀死Mdr1p过表达的多药耐药白色念珠菌.在用Mdr1p过表达的白色念珠菌小鼠感染模型中,小檗碱治疗组小鼠的平均存活时间(MST)得到了显著延长.本研究提示找寻药物外排泵的逆转剂可能为如何克服多药耐药问题提供了一个新的思路.

关 键 词：Candida albicans BERBERINE MULTIDRUG-RESISTANCE Drug excretion transporter MITOCHONDRIA 

分 类 号：R285.5[医药卫生—中药学]