糖尿病合并代谢相关脂肪性肝病的肠道菌群特征研究

doi:10.19871/j.cnki.xfcrbzz.2023.02.001

摘要/Abstract

摘要： 目的探讨糖尿病合并代谢相关脂肪性肝病（MAFLD）的发生发展与脂肪代谢相关基因及肠道菌群之间的相关性,为糖尿病合并MAFLD的防治提供了的新思路。方法 21只C57BL/6雄性小鼠随机分为糖尿病组（6只）和糖尿病合并MAFLD组（15只）,分别在糖尿病组小鼠和糖尿病合并MAFLD组小鼠给予高糖高脂饲料喂养35d和63d后,对两组小鼠血糖、血脂、肝功能、胰岛素、肝脏脂蛋白脂肪酶（LPl）和脂肪酸合成酶（Fasn）水平以及肠道菌群进行检测,并观察糖尿病合并MAFLD组小鼠肝脏HE染色及肠道紧密连接蛋白的变化情况。将2021年11月至2022年6月在沈阳市第六人民医院住院的15例糖尿病患者分为糖尿病组（7例）和糖尿病合并MAFLD组（8例）,后者又分为肥胖型（BMI>30kg/m²）4例和非肥胖型（BMI 8.5~22.9kg/m²）4例,对患者的粪便标本进行肠道菌群的宏基因检测。结果与糖尿病组小鼠相比,糖尿病合并MAFLD组小鼠的血脂、肝功能及胰岛素水平降低,血糖、肝脏LPl和Fasn水平升高,高密度脂蛋白胆固醇、LPl和Fasn组间比较有统计学差异（P<0.05）。两组小鼠造模成功前4周至造模成功后2周,肝脏HE染色及肠道紧密连接蛋白的变化显示,在MAFLD进展过程中肝细胞先出现脂肪堆积,随后发展为气球样变,肠道紧密连接蛋白的表达水平呈现逐渐下降趋势。糖尿病合并MAFLD的小鼠和患者中,瘤胃球菌属的丰度均较糖尿病组增高,组间比较有统计学差异（P<0.05）。与非肥胖型患者相比,肥胖型糖尿病合并MAFLD组患者的布劳特菌属、柯林斯菌属和瘤胃球菌属的丰度增加,双歧杆菌属的丰度降低,组间比较均有统计学差异（P<0.05）。结论在糖尿病合并MAFLD的发展过程中,肝脏的脂质沉积先于肠道紧密连接蛋白的改变而出现,瘤胃球菌属以及肝脏LPl、Fasn的高表达可能是MAFLD发生的重要因素。瘤胃球菌属、布劳特菌属和柯林斯菌属可能与肥胖相关,双歧杆菌属则对肥胖具有保护作用。

关键词: 糖尿病, 代谢相关脂肪性肝病, 肠道菌群, 肥胖

Abstract: Objective To explore the correlation between the occurrence and development of metabolic associated fatty liver disease (MAFLD) in diabetes and fat metabolism related genes and gut microbiota, and to provides a new idea for the prevention and treatment of diabetes combined with MAFLD. Method 21 Mice were randomly divided into diabetes group(6 cases) and diabetes with MAFLD group(15 cases). The mice in the diabetes group and the mice in the diabetes combined with MAFLD group were fed with high sugar and high fat diet for 35 days and 63 days respectively, then the levels of blood glucose, blood lipids, liver function, insulin, liver lipoprotein lipase (LPl), fatty acid synthase (Fasn) and intestinal flora in the two groups were detected, and the changes of liver HE staining and intestinal tight junction protein in diabetes with MAFLD group were observed. Fifteen diabetes patients hospitalized in the Sixth People's Hospital of Shenyang from November 2021 to June 2022 were divided into diabetes group (7 cases) and diabetes with MAFLD group (8 cases). Whole genome shotgun sequencing was used to detect the gut microbiota in feces samples of patients. Result Compared with the diabetes group, the levels of blood lipid, liver function and insulin in diabetes with MAFLD group were lower, while the levels of blood glucose, liver LPl and Fasn were higher. The HDL-C, LPl and Fasn had significant difference between the two groups (P<0.05). The changes in liver HE staining and intestinal tight junction protein were observed between 4 weeks before and 2 weeks after successful modeling in both groups of mice, with the progress of MAFLD, liver cells first appeared lipid accumulation, then developed into ballooning degeneration, and the expression levels of intestinal tight junction protein presented a gradual downward trend. Compared with the diabetes group, the Ruminococcus abundance in diabetes with MAFLD group was higher in both mice and patients, the difference was statistically significant (P<0.05). Compared with non obesity patients, the abundance of Ruminococcus, Blautia and Collinsella in obesity patients were increased, while the abundance of Bifidobacteria was decreased, the differences between the two groups were statistically significant(P<0.05). Conclusion In the development of diabetes combined with MAFLD, lipid deposition in the liver occurs before the changes of intestinal tight junction protein. Ruminococcus and high expression of LPl and Fasn in the liver may play an important role in the occurrence of MAFLD. Ruminococcus, Blautia and Collinsella may related to obesity, while Bifidobacterium has a protective effect on obesity.

Key words: Diabetes, Metabolic associated fatty liver disease, Gut microbiota, Obesity

中图分类号:

R587.1

刘星, 王晓梅, 谷野, 关欣, 张朕, 李欣悦, 李美雨希, 李立竹, 王岩. 糖尿病合并代谢相关脂肪性肝病的肠道菌群特征研究[J]. 新发传染病电子杂志, 2023, 8(2): 1-7.

Liu Xing, Wang Xiaomei, Gu Ye, Guan Xin, Zhang Zhen, Li Xinyue, Li Meiyuxi, Li Lizhu, Wang Yan. Study on the characteristics of gut microbiota in diabetes with metabolic associated fatty liver disease[J]. Electronic Journal of Emerging Infectious Diseases, 2023, 8(2): 1-7.

参考文献

[1] VERNON G, BARANOVA A, YOUNOSSI ZM, et al.Systematic review: The epidemiology and natural history of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis in adults[J]. Aliment Pharmacol Ther, 2011, 34(3): 274-285.
[2] 李玉丹, 董常峰, 刘映霞,等. VTQ与APRI诊断慢性乙型病毒性肝炎合并非酒精性脂肪肝肝纤维化的价值[J/CD]. 新发传染病电子杂志, 2017, 2(2):68-71,75.
[3] WU D, LIU Z, WANG Y, et al.Epigallocatechin-3-Gallate Alleviates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via Inhibition of Apoptosis and Promotion of Autophagy through the ROS/MAPK Signaling Pathway[J]. Oxid Med Cell Longev, 2021, 2021:5599997.
[4] 薛芮, 范建高. 代谢相关脂肪性肝病新定义的国际专家共识简介[J]. 临床肝胆病杂志, 2020, 36(6): 1224-1227.
[5] 范建高,曾静.非酒精性脂肪性肝病的流行现状与危害[J]. 中华消化杂志, 2020, 40(9): 577-580.
[6] 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]. Int J Mol Sci, 2019, 20(2): 395.
[7] TSAI HJ, TSAI YC, HUNG WW, et al.Gut Microbiota and Non-Alcoholic Fatty Liver Disease Severity in Type 2 Diabetes Patients[J]. J Pers Med, 2021, 11(3):238.
[8] 彭晓玲. 糖尿病合并MAFLD的临床特征及相关因素分析[D].大理:大理大学, 2022.
[9] TARGHER G, LONARDO A, BYRNE CD.Nonalcoholic fatty liver disease and chronic vascular complications of diabetes mellitus[J]. Nat Rev Endocrinol, 2018, 14(2):99-114.
[10] IRUZUBIETA P, MEDINA JM, FERNANDEZ-LOPEZ R, et al.A Role for Gut Microbiome Fermentative Pathways in Fatty Liver Disease Progression[J]. J Clin Med, 2020, 9(5): 1369-1373.
[11] ABU-SHANAB A, QUIGLEY EM.The role of the gut microbiota in nonalcoholic fatty liver disease[J]. Nat Rev Gastroenterol Hepatol, 2010, 7(12): 691-701.
[12] ALLIN KH, NIELSEN T, PEDERSEN O.Mechanisms in endocrinology: Gut microbiota in patients with type 2 diabetes mellitus[J]. Eur J Endocrinol, 2015, 172(4): 167-177.
[13] TAI N, WONG FS, WEN L, et al.The role of gut microbiota in the development of type 1, type 2 diabetes mellitus and obesity[J]. Rev Endocr Metab Disord, 2015, 16(1): 55-65.
[14] KERSTEN S.Physiological regulation of lipoprotein lipase[J]. Biochim Biophys Acta, 2014, 1841(7): 919-933.
[15] CHEN X, HUANG K, HU S, et al.FASN-Mediated Lipid Metabolism Regulates Goose Granulosa Cells Apoptosis and Steroidogenesis[J]. Front Physiol, 2020, 11: 600.
[16] 田丽艳, 范建高, 钱燕, 等. 链脲佐菌素糖尿病大鼠的肝脏损伤[J]. 肝脏, 2005, 10(2): 135-136.
[17] 谢璐蔓,黄葵,覃善芳.381例艾滋病患者肠道感染情况及耐药性分析[J/CD].新发传染病电子杂志,2021,6(3):220-224.
[18] DIAMANT M, BLAAK EE, DE VOS WM.Do nutrient-gut-microbiota interactions play a role in human obesity, insulin resistance and type 2 diabetes?[J]. Obes Rev, 2011, 12: 272-281
[19] LOOMBA R, SEGURITAN V, LI W, et al. Gut Microbiome-Based Metagenomic Signature for Non-invasive Detection of Advanced Fibrosis in Human Nonalcoholic Fatty Liver Disease[J]. Cell Metab, 2017, 25(5): 1054-1062.e5.
[20] GRABHERR F, GRANDER C, EFFENBERGER M, et al.Gut Dysfunction and Non-alcoholic Fatty Liver Disease[J]. Front Endocrinol (Lausanne), 2019, 10:611.
[21] BOURSIER J, MUELLER O, BARRET M, et al.The severity of nonalcoholic fatty liver disease is associated with gut dysbiosis and shift in the metabolic function of the gut microbiota[J]. Hepatology, 2016, 63(3):764-775.
[22] WANG B, JIANG X, CAO M, et al.Altered Fecal Microbiota Correlates with Liver Biochemistry in Nonobese Patients with Non-alcoholic Fatty Liver Disease[J]. Sci Rep, 2016, 6: 32002.
[23] SINHA SR, HAILESELASSIE Y, NGUYEN LP, et al. Dysbiosis-Induced Secondary Bile Acid Deficiency Promotes Intestinal Inflammation[J]. Cell Host Microbe, 2020, 27(4):659-670.e5.
[24] GOFFREDO M, MASS K, PARKS EJ, et al.Role of Gut Microbiota and Short Chain Fatty Acids in Modulating Energy Harvest and Fat Partitioning in Youth[J]. J Clin Endocrinol Metab, 2016,101(11): 4367-4376.
[25] TANG W, YAO X, XIA F, et al.Modulation of the Gut Microbiota in Rats by Hugan Qingzhi Tablets during the Treatment of High-Fat-Diet-Induced Nonalcoholic Fatty Liver Disease[J]. Oxid Med Cell Longev, 2018, 2018: 7261619.
[26] ZHANG X, SHEN D, FANG Z, et al.Human gut microbiota changes reveal the progression of glucose intolerance[J]. PLoS One, 2013, 8(8):e71108.
[27] RUIZ-LIMÓN P, MENA-VÁZQUEZ N, MORENO-INDIAS I, et al. Collinsella is associated with cumulative inflammatory burden in an established rheumatoid arthritis cohort[J]. Biomed Pharmacother, 2022, 153:113518.