摘要
目的 研究二氢杨梅素(dihydromyricetin,DHM)对代谢相关脂肪性肝病(metabolic-associated fatty liver disease,MAFLD)的改善作用,并揭示其调节肠黏膜屏障功能的作用机制。方法 32只雄性C57BL/6J小鼠随机分为4组(n=8):CON(对照,脂肪供能比为10%饮食)组、高脂饮食(HFD,脂肪供能比为45%)组、HFD+DHM组(添加质量比0.6%DHM的高脂饲料)和DHM组(添加质量比0.6%DHM的对照饲料),干预12周。每周测量小鼠体质量、摄食量;处死前进行空腹血糖、葡萄糖耐量和肠道通透性检测;试剂盒检测血清生化指标、脂多糖(LPS);肝组织进行油红O染色和HE染色,观察组织病理学变化;RT-qPCR和Western blot法检测肠上皮紧密连接蛋白、抗菌肽及肝脏炎症因子表达水平;流式细胞术检测结肠3型固有淋巴样细胞(group 3 innate lymphoid cells,ILC3)比例以及IL-22表达水平。结果 与CON组比较,HFD组小鼠体质量增加,糖耐量受损,血清总胆固醇、甘油三酯、谷丙转氨酶、谷草转氨酶升高,肝脏脂质沉积和炎性因子IL-1β、IL-6、TNF-α mRNA表达水平升高,肠道通透性和血清LPS增加,肠黏膜紧密连接蛋白ZO-1、Occludin及抗菌肽RegⅢβ、RegⅢγ表达水平降低,结肠ILC3及IL-22^(+)ILC3比例降低(P<0.05)。与HFD组比较,HFD+DHM组糖耐量受损明显改善,肝脏脂质沉积和炎性因子表达减少,血清LPS水平降低,肠紧密连接蛋白和抗菌肽表达水平增加,肠道通透性减少,结肠ILC3和IL-22^(+)ILC3比例增加(P<0.05)。结论 DHM可能通过改善肠黏膜屏障功能减少肝脏脂质堆积和炎症反应,改善MAFLD疾病进程。
Objective To investigate the ameliorative effects of dihydromyricetin(DHM) on metabolism-associated fatty liver disease(MAFLD),and to reveal the mechanism of its regulation of intestinal mucosal barrier function.Methods Thirty-two male C57BL/6J mice were randomly divided into 4 groups(n=8):CON(control diet,with 10% fat) group,HFD(high-fat diet,with 45% fat) group,HFD+DHM(high-fat diet+0.6% DHM) group,and DHM(control diet+0.6% DHM) group.All the subjects were intervened for 12 weeks,the body weight and food consumption of mice were measured weekly.Fasting blood glucose,glucose tolerance and intestinal permeability were tested before sacrifice.Serum levels of biochemical indexes and lipopolysaccharide(LPS) were detected with corresponding kits.The liver tissues were stained with HE and oil red O staining,respectively,and the histopathological changes were observed.Moreover,RT-qPCR and Western blotting were used to measure the expression levels of intestinal epithelial tight junction proteins,antimicrobial peptides and liver inflammatory factors.Flow cytometry was adopted to determine the proportion of group 3 innate lymphoid cells(ILC3) and the level of IL-22 in the colon.Results As compared with the CON group,the HFD group presented increased body weight,impaired glucose tolerance,as well as elevated serum total cholesterol,triglyceride,alanine transaminase and aspartate transaminase.HFD also led to liver lipid deposition and higher mRNA levels of IL-1β,IL-6 and TNF-α in the liver.The intestinal permeability and serum LPS were increased,whereas the expression levels of tight junction proteins ZO-1 and Occludin and antimicrobial peptides RegⅢ β and RegⅢ γ in the intestinal mucosa were decreased,and the proportions of ILC3 and IL-22^(+) ILC3 in the colon were declined(P<0.05).In the HFD+DHM group,the impaired glucose tolerance was significantly improved,the hepatic lipid deposition and the expression of inflammatory factors were reduced,so was the serum LPS level.DHM also upregulated the levels of intestinal
作者
蒋玲
李鹏飞
候鹏飞
周杰
董妞
张乾勇
易龙
糜漫天
JIANG Ling;LI Pengfei;HOU Pengfei;ZHOU Jie;DONG Niu;ZHANG Qianyong;YI Long;MI Mantian(Research Center for Army Nutrition and Food Safety,Chongqing Key Laboratory of Nutrition and Food Safety,Chongqing Medical Nutrition Research Center,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2023年第2期146-154,共9页
Journal of Army Medical University
基金
国家自然科学基金面上项目(81872625、82173505)。
关键词
二氢杨梅素
代谢相关脂肪性肝病
肠黏膜屏障
肠上皮细胞
3型固有淋巴样细胞
dihydromyricetin
metabolism-related fatty liver disease
intestinal mucosal barrier
intestinal epithelial cells
group 3 innate lymphoid cells
