鸠坑龙井茶对高脂饮食C57BL/6小鼠肝脂肪变性SREBPs通路信号的影响及肠道菌群调节作用研究

doi:10.13305/j.cnki.jts.2023.04.010

茶叶科学 ›› 2023, Vol. 43 ›› Issue (4): 576-592.doi: 10.13305/j.cnki.jts.2023.04.010

• 研究报告 • 上一篇

鸠坑龙井茶对高脂饮食C57BL/6小鼠肝脂肪变性SREBPs通路信号的影响及肠道菌群调节作用研究

龚明秀^1,2, 袁懿炜^1,2, 张一帆^1,2, 叶江成^1,2, 郭丽³, 李晓军⁴, 黄皓⁴, 毛宇骁⁵, 赵芸⁵, 赵进^1,2,*

1.中国计量大学生命科学学院,食品营养与质量安全研究所,浙江杭州 310018;
2.特色农产品品质与危害物控制技术浙江省重点实验室,浙江杭州 310018;
3.中国农业科学院茶叶研究所,浙江杭州 310008;
4.浙江艺福堂茶业有限公司博士创新工作站,浙江杭州 311500;
5.杭州市农业科学研究院,浙江杭州 310024

收稿日期:2023-01-16 修回日期:2023-04-12 出版日期:2023-08-15 发布日期:2023-08-24
通讯作者: *zhaojin@cjlu.edu.cn
作者简介:龚明秀,女,硕士研究生,主要从事药食同源植物营养与功效评价研究。
基金资助:
浙江省重点研发计划项目（2020C02045）、杭州市农业与社会发展科研项目（202203A06、202203A11、202203B09）、开化茶产业提升浙江省团队科技特派员项目、杭州市科技特派员项目（20221122I80）、衢州市重点科技攻关项目科技强农专项（2023k098）

Effect of Jiukeng Longjing Tea on SREBPs Signaling Pathway and Gut Microbiota Regulation in High-fat Diet C57BL/6 Mice with Hepatic Steatosis

GONG Mingxiu^1,2, YUAN Yiwei^1,2, ZHANG Yifan^1,2, YE Jiangcheng^1,2, GUO Li³, LI Xiaojun⁴, HUANG Hao⁴, MAO Yuxiao⁵, ZHAO Yun⁵, ZHAO Jin^1,2,*

1. Institute of Food Nutrition and Quality Safety, College of Life Sciences, China Jiliang University, Hangzhou 310018, China;
2. Key Laboratory of Pecialty Agri-product Quality and Hazard Controlling Technology of Zhejiang Province, Hangzhou 310018, China;
3. Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
4. Doctor Innovation Workstation of Zhejiang Yifutang Tea Industry Co., Ltd., Hangzhou 311500, China;
5. Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China

Received:2023-01-16 Revised:2023-04-12 Online:2023-08-15 Published:2023-08-24

摘要/Abstract

摘要： 探究鸠坑龙井茶水提物（LJT）对小鼠肝组织脂质代谢SREBPs通路信号影响及肠道菌群的调节作用。通过高脂饮食诱导小鼠构建非酒精性脂肪肝（NAFL）模型,并给予LJT（300 mg·kg^-1）灌胃干预。定期记录小鼠的体质量,检测小鼠血清生化指标和葡萄糖耐受水平,观察并分析Hematoxylin-Eosin（HE）染色、油红O染色肝组织切片特征;应用Real-time qPCR技术检测小鼠肝组织SREBPs通路7个基因（SREBP-1c、FAS、SCD-1、ACC-1、SREBP-2、HMGCR、PPARγ）的相对表达量,采用蛋白免疫印记技术（Western Blot）分析肝组织蛋白质表达水平,同时对小鼠肠道菌群进行高通量测序（16 S rDNA）并分析其结构。结果显示,LJT干预后小鼠体质量、血糖AUC、血清TG、TC、LDL-C和肝脏中TG、TC水平有显著下降,龙井组小鼠肝组织SREBP-1c、FAS、ACC-1、SCD-1和PPARγ蛋白表达水平降低,SREBP-1c、SCD-1、FAS、ACC-1、SREBP-2、HMGCR、PPARγ基因的相对表达量显著下调;16 S rDNA分析发现,小鼠肠道菌群门水平主要为Firmicutes、Bacteroidota、Desulfobacterota和Actinobacteriota 4类,LJT有效延缓了高脂饮食引起的Firmicutes相对丰度升高和Bacteroidota相对丰度下降趋势,并增加了肠道菌群的物种丰度。结果表明,LJT能够干预小鼠肝脂肪变性SREBPs通路信号表达,改善小鼠肠道菌群紊乱,具有降脂减肥作用。

关键词: 鸠坑龙井茶, 非酒精性脂肪肝, SREBPs通路, 肠道菌群紊乱, 降脂作用

Abstract: To investigate the effect of Jiukeng Longjing tea water extract (LJT) on liver steatosis and the regulation of gut microbiota in C57BL/6 mice fed with high-fat diet, a non-alcoholic fatty liver model was established in mice induced by a high-fat diet, and LJT (300 mg·kg^-1) was gavaged for intervention. The body weight of mice was recorded regularly, and serum biochemical indicators such as AST, ALT, TC, TG, LDL-C, HDL-C, and glucose tolerance levels were measured. The characteristics of HE staining and oil red O staining liver tissue sections were observed and analyzed. Real-time qPCR technology was used to detect the expressions of seven genes including SREBP-1c, FAS, SCD-1, ACC-1, SREBP-2, HMGCR, and PPARγ in mouse liver tissues. The relative expressions of proteins related to lipid metabolism were studied by western blot. At the same time, the gut microbiota of mice was sequenced by high-throughput sequencing (16 S rDNA) and its structure was analyzed. The results show that the body weight, blood glucose AUC, serum TG, TC, LDL-C, and liver TG, TC levels significantly decreased under LJT intervention. Western blot shows that LJT intervention reduced the expressions of SREBP-1c, FAS, ACC-1, SCD-1, and PPARγ in liver tissue of mice. LJT also significantly downregulated the relative expressions of SREBP-1c, SCD-1, FAS, ACC-1, SREBP-2, HMGCR and PPARγ in liver tissue. The 16 S rDNA detection reveals that the levels of gut microbiota were mainly classified into four categories: Firmicutes, Bacteroidota, Desulfobacterota, and Actinobaciota. LJT could effectively alleviate the trend of increasing the relative abundance of Firmicutes and decreasing the relative abundance of Bacteroidota caused by high-fat diet, and increase the species abundance of gut microbiota. Therefore, LJT could interfere with the signal expression of SREBPs pathway in mouse liver steatosis, and improve the disturbance of gut microbiota in mice, thereby achieve the effect of reducing fat and weight loss.

Key words: Jiukeng Longjing tea, non-alcoholic fatty liver, SREBPs path, gut microbiota dysbiosis, lipid-lowering effect

中图分类号:

龚明秀, 袁懿炜, 张一帆, 叶江成, 郭丽, 李晓军, 黄皓, 毛宇骁, 赵芸, 赵进. 鸠坑龙井茶对高脂饮食C57BL/6小鼠肝脂肪变性SREBPs通路信号的影响及肠道菌群调节作用研究[J]. 茶叶科学, 2023, 43(4): 576-592. doi: 10.13305/j.cnki.jts.2023.04.010.

GONG Mingxiu, YUAN Yiwei, ZHANG Yifan, YE Jiangcheng, GUO Li, LI Xiaojun, HUANG Hao, MAO Yuxiao, ZHAO Yun, ZHAO Jin. Effect of Jiukeng Longjing Tea on SREBPs Signaling Pathway and Gut Microbiota Regulation in High-fat Diet C57BL/6 Mice with Hepatic Steatosis[J]. Journal of Tea Science, 2023, 43(4): 576-592. doi: 10.13305/j.cnki.jts.2023.04.010.

参考文献

[1] Hardy T, Oakley F, Anstee Q M, et al.Nonalcoholic fatty liver disease: pathogenesis and disease spectrum[J]. Annual Review of Pathology, 2016, 11: 451-496.
[2] Paternostro R, Trauner M.Current treatment of non-alcoholic fatty liver disease[J]. Journal of Internal Medicine, 2022, 292(2): 190-204.
[3] Brouwers M C G J, Simons N, Stehouwer C D A, et al. Non-alcoholic fatty liver disease and cardiovascular disease: assessing the evidence for causality[J]. Diabetologia, 2020, 63(2): 253-260.
[4] Carmiel-Haggai M, Cederbaum A I, Nieto N.A high-fat diet leads to the progression of non-alcoholic fatty liver disease in obese rats[J]. The FASEB Journal, 2005, 19(1): 136-138.
[5] El-Koofy N M, Anwar G M, El-Raziky M S, et al. The association of metabolic syndrome, insulin resistance and non-alcoholic fatty liver disease in overweight/obese children[J]. Saudi Journal of Gastroenterology, 2012, 18(1): 44-49.
[6] Powell E E, Wong V W, Rinella M.Non-alcoholic fatty liver disease[J]. Lancet, 2021, 397(10290): 2212-2224.
[7] Bashiardes S, Shapiro H, Rozin S, et al.Non-alcoholic fatty liver and the gut microbiota[J]. Molecular Metabolism, 2016, 5(9): 782-794.
[8] Charroux B, Royet J.Gut-derived peptidoglycan remotely inhibits bacteria dependent activation of SREBP by Drosophila adipocytes[J]. Plos Genetics, 2022, 18(3): e1010098. doi: 10.1371/journal.pgen.1010098.
[9] 朱荫, 邵晨阳, 张悦, 等. 不同茶树品种龙井茶香气成分差异分析[J]. 食品工业科技, 2018, 39(23): 241-246, 254.
Zhu Y, Shao C Y, Zhang Y, et al.Comparison of differences in aroma constituents of Longjing tea produced from different tea germplasms[J]. Science and Technology of Food Industry, 2018, 39(23): 241-246, 254.
[10] 孙达, 龚恕, 崔宏春, 等. 不同品种茶树春秋季鲜叶超微绿茶粉适制性研究[J]. 浙江农业学报, 2021, 33(3): 437-446.
Sun D, Gong S, Cui H C, et al.Suitablity of fresh spring and autumn leaves from different tea cultivars for ultramicro green tea powder production[J]. Acta Agriculturae Zhejiangensis, 2021, 33(3): 437-446.
[11] 王素敏, 徐欢欢, 黄业伟, 等. 茶多酚的降脂作用及其机制研究进展[J]. 食品研究与开发, 2016, 37(10): 219-224.
Wang S M, Xu H H, Huang Y W, et al.Review on the effect of tea polyphenols in hypolipidemic and its medchanism[J]. Food Research and Development, 2016, 37(10): 219-224.
[12] Li B Y, Li H Y, Zhou D D, et al.Effects of different green tea extracts on chronic alcohol induced-fatty liver disease by ameliorating oxidative stress and inflammation in mice[J]. Oxid Med Cell Longev, 2021, 2021: 5188205. doi: 10.1155/2021/5188205.
[13] Ma H, Zhang B, Hu Y, et al.The novel intervention effect of cold green tea beverage on high-fat diet induced obesity in mice[J]. Journal of Functional Foods, 2020, 75: 104279. doi: 10.1016/j.jff.2020.104279.
[14] 王忠民, 吴谋成, 李小定, 等. 葡萄多糖的提取及含量测定[J]. 新疆农业大学学报, 2002, 25(2): 57-58.
Wang Z M, Wu M C, Li X D, et al.Extraction and contents, measurement of the VLP[J]. Journal of Xinjiang Agricultural University, 2002, 25(2): 57-58.
[15] Li M, Xu J, Zhang Y, et al.Comparative analysis of fecal metabolite profiles in HFD-induced obese mice after oral administration of Huangjinya green tea extract[J]. Food and Chemical Toxicology, 2020, 145: 111744. doi: 10.1016/j.fct.2020.111744.
[16] 冯琳, 龚自明, 刘盼盼, 等. 青砖毛茶对高脂饮食小鼠肠道微生物的影响[J]. 中国食品学报, 2021, 21(7): 87-96.
Feng L, Gong Z M, Liu P P, et al.Effects of Qingzhuan Maocha on gut microbiota in high-fat diet fed mice[J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(7): 87-96.
[17] Ma H, Zhang B, Hu Y, et al.Correlation analysis of intestinal redox state with the gut microbiota reveals the positive intervention of tea polyphenols on hyperlipidemia in high fat diet fed mice[J]. Journal of Agricultural and Food Chemistry, 2019, 67(26): 7325-7335.
[18] Velázquez K T, Enos R T, Bader J E, et al.Prolonged high-fat-diet feeding promotes non-alcoholic fatty liver disease and alters gut microbiota in mice[J]. World Journal of Hepatology, 2019, 11(8): 619-637.
[19] Aydos L R, Do Amaral L A, De Souza R S, et al. Nonalcoholic fatty liver disease induced by high-fat diet in C57BL/6 models[J]. Nutrients, 2019, 11(12): 3067. doi: 10.3390/nu11123067.
[20] Yin J, Li Y, Han H, et al.Melatonin reprogramming of gut microbiota improves lipid dysmetabolism in high-fat diet-fed mice[J]. Journal of Pineal Research, 2018, 65(4): e12524. doi: 10.1111/jpi.12524.
[21] 黄玉晶. 邻苯二甲酸酯母体暴露对早产和胎儿发育的影响及过氧化物酶体增殖物激活受体在其中的作用研究[D]. 重庆: 第三军医大学, 2014.
Huang Y J.The role of peroxisome proliferator activated receptor in the effects of gestation exposure to phthalates on preterm delivery and fetal development [D]. Chongqing: Army Medical University, 2014.
[22] 刘亚茹, 苗志国, 高明磊, 等. PPARγ在动物脂肪发育中的研究进展[J]. 黑龙江畜牧兽医, 2019(1): 32-35.
Liu Y R, Miao Z G, Gao M L, et al.Research advance on PPARγ in animal adipose tissue[J]. Heilongjiang Animal Science and Veterinary Medicine, 2019(1): 32-35.
[23] Shi W, Hou T, Guo D, et al.Evaluation of hypolipidemic peptide (Val-Phe-Val-Arg-Asn) virtual screened from chickpea peptides by pharmacophore model in high-fat diet-induced obese rat[J]. Journal of Functional Foods, 2019, 54: 136-145.
[24] Geng T T, Liu Y, Xu Y T, et al.H19 lncRNA promotes skeletal muscle insulin sensitivity in part by targeting AMPK[J]. Diabetes, 2018, 67(11) : 2183-2198.
[25] Loregger A, Raaben M, Nieuwenhuis J, et al.Haploid genetic screens identify SPRING/C12ORF49 as a determinant of SREBP signaling and cholesterol metabolism[J]. Nature Communications, 2020, 11: 1128. doi: 10.1038/s41467-020-14811-1.
[26] Wei S, Espenshade P J.Expanding roles for SREBP in metabolism[J]. Cell Metabolism, 2012, 16(4): 414-419.
[27] Milosevic I, Vujovic A, Barac A, et al.Gut-liver axis, gut microbiota, and its modulation in the management of liver diseases: a review of the literature[J]. International Journal of Molecular Sciences, 2019, 20(2): 395. doi: 10.3390/ijms20020395.
[28] Soderborg T K, Clark S E, Mulligan C E, et al.The gut microbiota in infants of obese mothers increases inflammation and susceptibility to NAFLD[J]. Nature Communications, 2018, 9: 4462. doi: 10.1038/s41467-018-06929-0.
[29] Ley R E, Bäckhed F, Turnbaugh P, et al.Obesity alters gut microbial ecology[J]. PNAS, 2005, 102(31): 11070-11075.
[30] Do M H, Lee H B, Oh M J, et al.Polysaccharide fraction from greens of Raphanus sativus alleviates high fat diet-induced obesity[J]. Food Chemistry, 2021, 33:128395. doi: 10.1016/j.foodchem.2020.128395.
[31] Guo J L, Han X, Zhan J C, et al.Vanillin alleviates high fat diet-induced obesity and improves the gut microbiota composition[J]. Frontiers in Microbiology, 2018, 9: 2733. doi: 10.3389/fmicb.2018.02733.
[32] Beaumont M, Andriamihaja M, Lan A, et al.Detrimental effects for colonocytes of an increased exposure to luminal hydrogen sulfide: the adaptive response[J]. Free Radical Biology and Medicine, 2016, 93: 155-164.
[33] Carter J K, Bhattacharya D, Borgerding J N, et al.Modeling dysbiosis of human NASH in mice: loss of gut microbiome diversity and overgrowth of Erysipelotrichales[J]. Plos One, 2021, 16(1): e0244763.
[34] Smith B J, Miller R A, Ericsson A C, et al.Changes in the gut microbiome and fermentation products concurrent with enhanced longevity in acarbose-treated mice[J]. BMC Microbiology, 2019, 19: 130. doi: 10.1186/s12866-019-1494-7.
[35] Musso G, Gambino R, Cassader M.Obesity, diabetes, and gut microbiota: the hygiene hypothesis expanded?[J]. Diabetes Care, 2010, 33(10): 2277-2284.
[36] Den Besten G, Bleeker A, Gerding A, et al.Short-chain fatty acids protect against high-fat diet-induced obesity via a PPARγ-dependent switch from lipogenesis to fat oxidation[J]. Diabetes, 2015, 64(7): 2398-2408.

鸠坑龙井茶对高脂饮食C57BL/6小鼠肝脂肪变性SREBPs通路信号的影响及肠道菌群调节作用研究

Effect of Jiukeng Longjing Tea on SREBPs Signaling Pathway and Gut Microbiota Regulation in High-fat Diet C57BL/6 Mice with Hepatic Steatosis

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