St cancer, and anti-obesity and hepatoprotective effects [204]. Also, it has been located that these biological activities are mostly attributed towards the big functional elements, such as polyphenols [25]. Depending on the various fermentation mTORC2 Inhibitor medchemexpress degrees and processes, tea can be normally classified into six important sorts, such as green tea, MEK1 Inhibitor Biological Activity yellow tea, white tea, oolong tea, black tea, and dark tea [26]. The green tea, yellow tea, and white tea were un-fermented or slight-fermented. The oolong tea (semi-fermented), black tea (deep-fermented) and dark tea (post-fermented) have received increasing focus on the regulation of gut microbiota in recent years, since a lot of microbes and metabolites were created in the fermented approach [270]. For instance, a earlier study has reported that black and oolong teas could substantially alter the abundance of Bacteroides, Parabacteroides, and Akkermansia in mice fed using a high-fat diet plan (HFD) [31]. Similarly, a further research has shown that Fuzhuan Brick Tea, a single dark tea, could alter the structure of gut microbiota community in HFD-fed mice, with a significant boost within the abundance of Bifidobacteriaceae [32]. Having said that, the potential part of gut microbiota modulated by oolong tea, black tea and dark tea within the prevention and management of AFLD continues to be not clearly understood. Thus, in the present study, our aim is always to discover and investigate the effects of oolong tea, black tea, and dark tea on AFLD in chronic alcohol-exposed mice as well as the prospective role of gut microbiota in mediating the effects of these teas on AFLD. two. Supplies and Techniques two.1. Preparation of Tea Extracts The distinct information and facts regarding the two black teas, two oolong teas, and two dark teas purchased from China is shown in Table 1. The preparation strategy on the tea extracts was as follows. Briefly, the tea sample (10 g) was extracted 3 times with 100 mL of boiling water in water bath shaker-DKZ-450B (98 C) (Sensin, Shanghai, China) for 10 min. Subsequently, all extracted options were combined and filtered, and then concentrated viaFoods 2021, 10,three ofa rotary evaporator (R-501, Cancun, Shanghai, China) below a vacuum environment. Lastly, the concentrated filtrate from tea extract was freeze-dried by way of a freeze dryer (Labconco7752001, Kansas City, MO, USA) to obtain a powder, which was stored at -80 C.Table 1. The specific facts of six selected teas. Number BT1 BT2 OT1 OT2 DT1 DT2 Category Black Tea Black Tea Oolong Tea Oolong Tea Dark Tea Dark Tea Name Dianhong Tea Yingde Black Tea Tieguanyin Tea Fenghuang Danzong Tea Fuzhuan Brick tea Selenium-Enriched Dark Tea Production Spot Xishuangbanna, Yunnan Yingde, Guangdong Xiamen, Fujian Shantou, Guangdong Changsha, Hunan Enshi, Hubei Fermentation Degree Deep-fermented Deep-fermented Semi-fermented Semi-fermented Post-fermented Post-fermented2.2. Experimental Animals and Feeding The 8-week-old C57BL/6J male mice had been applied within this study, which had been purchased from the Guangdong Healthcare Laboratory Animal Center (Guangzhou, China). All mice have been fed in specific-pathogen-free (SPF) laboratory animal room and they could consume and drink freely. On top of that, the animal space was maintained beneath the light (12 h lightdark cycle), the relative humidity (400 ) and also the temperature (22 0.five C) controlled environment. In addition, the AFLD mouse models had been induced by the modified LieberDeCarli ethanol liquid diet regime (TP 4030B), and animals within the handle group had been fed.