Puerarin prevents high-fat diet-induced obesity by enriching Akkermansia muciniphila in the gut microbiota of mice

RESEARCH ARTICLE Puerarin prevents high-fat diet-induced obesity by enriching Akkermansia muciniphila in the gut microbiota of mice Lei Wang, Yongzheng Wu, Lingjia Zhuang, Xiufang Chen, Haiyan Min, Shiyu Song, Qiao Liang, An-Dong Li*, Qian Gao ID* Jiangsu Key Laboratory of Molecular Medicine and Center for Translational Medicine, Medical School of Nanjing University, Nanjing, Jiangsu Province, China a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 OPEN ACCESS Citation: Wang L, Wu Y, Zhuang L, Chen X, Min H, Song S, et al. (2019) Puerarin prevents high-fat diet-induced obesity by enriching Akkermansia muciniphila in the gut microbiota of mice. PLoS ONE 14(6): e0218490. https://doi.org/10.1371/ journal.pone.0218490 Editor: Vanessa Souza-Mello, State University of Rio de Janeiro, BRAZIL Received: March 7, 2019 Accepted: June 2, 2019 * (QG); (ADL) Abstract Growing evidence indicates that the gut microbiota plays a significant role in the pathophysiological processes of obesity and its related metabolic symptoms in the host. Puerarin, an active ingredient in the root of Pueraria lobate has been suggested to have a potent antiobesity effect. Herein, we tested whether this effect of puerarin is associated with changes in the gut microbiota. In addition to reducing body weight, inflammation, and insulin resistance, puerarin administration significantly altered the composition of the gut microbiota. Notably, puerarin treatment greatly increased the abundance of Akkermansia muciniphila, a mucin-degrading bacterium known to be beneficial for host metabolism and significantly downregulated in high-fat diet–fed mice. Further experiments revealed that puerarin increased intestinal expression levels of Muc2 and Reg3g and protected intestinal barrier function (normal permeability) by increasing the expression of ZO-1 and occludin in vivo and in vitro. These data suggest that puerarin’s enriching effect on A. muciniphila is mediated, at least in part, by a host cellular response to protect the host from diet-induced metabolic disorders and other diseases. Published: June 24, 2019 Copyright: © 2019 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: The sequence data supporting the results of this article are available in the NCBI under the SRA accession number PRJNA510293. Funding: The authors gratefully acknowledge the financial support of the Key Project from the National Natural Science Foundation of China (81570775). Competing interests: The authors have declared that no competing interests exist. Introduction Obesity as a common precursor of many chronic diseases is associated with low-grade chronic inflammation and intestinal dysbiosis [1, 2]. It can lead to various metabolism-related problems including cardio- and cerebrovascular diseases, type 2 diabetes, chronic kidney disease, musculoskeletal disorders, and digestive diseases [3, 4], therefore represents a major health problem. Studies have revealed that the gut microbiota plays a crucial role in diet-related obesity and glucose and/or lipid metabolism disorders [5, 6]. For example, “obesity-related” and “normal-weight-related” microbiotas exhibit significant differences in their composition and profile and in their ability to utilize glucose and/or lipids in humans and other animals [7, 8]. Moreover, germ-free animals show a significant increase in body weight and metabolic dysfunctions after receiving gut microbiota isolated from donors with high-fat diet (HFD) induced obesity [9], suggesting a causal relation between gut dysbiosis and obesity. The gut PLOS ONE | https://doi.org/10.1371/journal.pone.0218490 June 24, 2019 1 / 19 Puerarin changes gut microbiota in obese mice microbiota as one of the predisposing factors of metabolic disorders affects the expression of host genes that regulate energy expenditure and storage especially in livers as the intestinal microbiota determines the development of non-alcoholic fatty liver disease in mice [10, 11], whereas, steatosis is among the most common phenotype of HFD mice [12]. More recently, studies revealed that the specific bacterial species Akkermansia muciniphila (A. muciniphila) inversely correlates with obesity and diabetes in both mice and humans [13]. Thus, the gut microbiota may be crucial for metabolic disease etiology. In fact, the gut microbiota has been suggested as a therapeutic target in various diseases, including obesity and diabetes [14]. Currently, most efforts are focused on fecal microbiota transplantation with encouraging results [15]. Nonetheless, concerns have been raised regarding its safety both at the biological and operative levels [16]. Moreover, the inconsistency in fecal microbiota compositions among donors as well as differences in the procedures are an unavoidable issue [17], in addition to the unregulated state of fecal microbiota transplantation, outside the approved clinical practice [18]. Thus, a simpler and measurable approach, e.g., using food or chemical compounds that are effective and safe for regulating the gut microbiota is desirable. Puerarin is the major bioactive ingredient in the root of Pueraria lobate (Wild) Ohwi and has been known as a combination of medicine and food in traditional Chinese medicine for thousands of years [19]. Puerarin possesses a wide spectrum of biological activities including an anti-obesity effect and has been used in alternative medicine for the treatment of various metabolic disorders [20–22]. Nevertheless, the mechanisms of action of puerarin in these diseases remain elusive. Herein, we investigated the effect of puerarin on the gut microbiota in mice with HFDinduced obesity. The aim of this study was to establish a link between puerarin’s anti-obesity effect and gut dysbiosis, and to identify the puerarin-induced specific microbiota which may help suppressing obesity. Our results suggest that puerarin is a potent therapeutic agent for gut dysbiosis and provide new evidence for understanding the involvement of the gut microbiota in HFD-induced metabolic disorders. Materials and methods Ethics statement Animal experiments were carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals (Nanjing University) and were approved by the Institutional Animal Care and Use Committee of Nanjing University. Animals and experimental design Male C57BL/6 mice (6–7 weeks old, n = 24) were purchased for this study from the Model Animal Research Center of Nanjing University. Prior to our experiments, the animals were provided with feed and distilled water ad libitum. They were housed at a facility with a good ventilation and air filtration system at 20 ± 2˚C, 50% ± 10% relative humidity, and 12 h light/ dark cycle. All pr (...truncated)