辣椒素保护肠黏膜屏障改善酒精性肝病小鼠肝损伤的实验研究
Experimental study on capsaicin improving liver damage in mice with alcoholic liver disease by protecting intestinal mucosal barrier
- 上海中医药杂志 2022年56卷第5期 页码：54-59
- 作者机构：
1.上海中医药大学附属普陀医院感染科肝病实验室（上海 200062）
2.上海中医药大学附属普陀医院中心实验室（上海 200062）
- 作者简介：
马文婷，女，硕士，副主任医师，主要从事中医药防治慢性肝病的研究工作
刘成，研究员，硕士研究生导师； E-mail：liucheng0082010@163.com
- 基金信息：国家自然科学基金项目(81803232;81873136;81803898;82004106);上海市卫健委卫生行业临床研究专项(202040149);上海市普陀区卫生系统自主创新科学基金项目(ptkwws201904);上海市普陀区卫生健康系统科技创新项目(ptkwws202223)
DOI：10.16305/j.1007-1334.2022.2111131
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马文婷,陶乐,刘旭凌等.辣椒素保护肠黏膜屏障改善酒精性肝病小鼠肝损伤的实验研究[J].上海中医药杂志,2022,56(05):54-59.
MA Wenting,TAO Le,LIU Xuling,et al.Experimental study on capsaicin improving liver damage in mice with alcoholic liver disease by protecting intestinal mucosal barrier[J].Shanghai Journal of Traditional Chinese Medicine,2022,56(05):54-59.
马文婷,陶乐,刘旭凌等.辣椒素保护肠黏膜屏障改善酒精性肝病小鼠肝损伤的实验研究[J].上海中医药杂志,2022,56(05):54-59. DOI： 10.16305/j.1007-1334.2022.2111131.
MA Wenting,TAO Le,LIU Xuling,et al.Experimental study on capsaicin improving liver damage in mice with alcoholic liver disease by protecting intestinal mucosal barrier[J].Shanghai Journal of Traditional Chinese Medicine,2022,56(05):54-59. DOI： 10.16305/j.1007-1334.2022.2111131.
摘要
目的,2,研究辣椒素对酒精性肝病小鼠肝脏和肠黏膜病变的改善作用，并探讨其作用机制。,方法,2,24只雄性C57/BL6小鼠随机分为对照组、乙醇组和乙醇辣椒素组，乙醇组及乙醇辣椒素组采用4周慢性乙醇液体饲料喂养加急性乙醇灌胃法复制酒精性肝病小鼠模型，对照组给予等量液体饲料。造模第3周起，乙醇辣椒素组给予辣椒素（10 mg/kg）灌胃，隔日1次，每周3次，其余两组给予等体积蒸馏水灌胃。造模4周结束后，乙醇组及乙醇辣椒素组给予31.5%乙醇灌胃，对照组给予45%糊精灌胃，9 h 后处死小鼠，留取血、肝脏、小肠组织样本。检测血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和三酰甘油（TG）含量；苏木精-伊红（HE）染色观察肝脏和小肠黏膜病理变化，油红染色观察肝脏脂质沉积情况。采用RT-PCR检测肝脏炎症因子肿瘤坏死因子-α（,TNF,-,α,）、白介素-6（,IL,-,6,）、白介素-1β（,IL,-,1β,）及小肠组织炎症因子,TNF,-,α,、,IL,-,6,、单核细胞趋化蛋白-1（,MCP,-,1,）及紧密连接蛋白中闭合小环蛋白-1（,ZO,-,1,）、咬合蛋白（,occludin,）、闭合蛋白-1（,claudin,-,1,）的mRNA表达水平。,结果,2,与对照组比较，乙醇组小鼠ALT、AST和TG水平显著升高（,P,<,0.05）；肝脏组织中,TNF,-,α,、,IL,-,6,、,IL,-,1β ,mRNA均显著升高（,P,<,0.05）。HE染色可见乙醇组小鼠肝脏脂肪变性及炎细胞浸润明显，油红染色显示乙醇组小鼠肝细胞内有大量脂滴。小肠HE染色可见乙醇组小肠绒毛高度缩短，上皮细胞水肿，黏膜下炎细胞浸润增多。乙醇组小鼠小肠组织,TNF,-,α,、,IL,-,6,、,MCP,-,1 ,mRNA水平较对照组显著升高（,P,<,0.05），而紧密连接蛋白的,ZO,-,1,、,occludin,、,claudin,-,1 ,mRNA水平则较对照组显著降低（,P,<,0.01）。与乙醇组比较，乙醇辣椒素组小鼠ALT、AST和TG水平显著减低（,P,<,0.05），肝脏组织中,TNF,-,α,、,IL,-,6,、,IL,-,1β ,mRNA均显著减低（,P,<,0.05），肝脏脂肪变及肠黏膜病理损伤均减轻，肠黏膜,TNF,-,α,、,IL,-,6,、,MCP,-,1 ,mRNA水平明显降低（,P,<,0.05），而紧密连接蛋白的,ZO,-,1,、,occludin,、,claudin,-,1 ,mRNA水平显著上调（,P,<,0.01）。,结论,2,辣椒素对酒精性肝病小鼠肝脏有保护作用，其作用机制可能与保护肠黏膜屏障、减轻肠渗漏有关。
Abstract
Objective,2,To study the effect of capsaicin on the improvement of liver and intestinal mucosal lesions in mice with alcoholic liver disease （ALD）， and explore its related mechanisms.,Methods,2,Twenty-four male C57/BL6 mice were randomly divided into the control group， the alcohol group and the alcohol capsaicin group. The mice in the alcohol group and the alcohol capsaicin group were fed with ethanol liquid diet for 4 weeks and gavaged with alcohol to replicate the alcoholic liver disease model. The control group was administered with isocaloric liquid feed. From the 3rd week of modeling， the alcohol capsaicin group was given capsaicin （10 mg/kg body weight） by gavage every other day and three times a week， while the other two groups were given an equal volume of distilled water by gavage. After 4 weeks of modeling， mice in the alcohol group and the alcohol capsaicin group were given 31.5% ethanol by gavage， while the mice in the control group were given 45% dextrin by gavage. The mice were sacrificed 9 hours later， and blood， liver and small intestine tissue samples were collected. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST） and triglyceride （TG） levels were detected by enzyme-linked immunosorbent assay. The pathological changes of liver and small intestine mucosa were observed after HE staining. The lipid deposition in the liver was observed by oil red O staining. RT-PCR was used to detect the mRNA expression levels of liver inflammatory factors including tumor necrosis factor-α （,TNF,-,α,）， interleukin 6 （,IL,-,6,）， interleukin 1β （,IL,-,1β,）， small intestinal tissue inflammatory factors including ,TNF,-,α,， ,IL,-,6,， monocyte chemoattractant protein 1 （,MCP,-,1,） and tight junction proteins including ,ZO,-,1,， ,occludin, and ,claudin,-,1,.,Results,2,Compared with the control group， the serum ALT， AST， TG levels and liver mRNA expressions of ,TNF,-,α,， ,IL,-,6,， ,IL,-,1β, were significantly increased in the alcohol group （,P,<,0.05）. HE staining showed that hepatic steatosis and inflammatory cells in the alcohol group was obvious. Oil red staining showed a large number of lipid droplets in the liver cells of the alcohol group. Shortened small intestine villi， epithelial cell edema and increased submucosal inflammatory cell infiltration in the alcohol group were observed by HE staining of small intestine mucosa. Compared with those in the control group， mRNA relative expression of ,TNF,-,α,， ,IL,-,6, and ,MCP,-,1, in the small intestine tissue of mice were increased significantly （,P,<,0.05）， while mRNA relative expression of tight junction protein ,ZO,-,1,， ,occludin, and ,claudin,-,1, were significantly decreased （,P,<,0.01）. Compared with those in the alcohol group， the serum ALT， AST， TG levels and liver mRNA expressions of ,TNF,-,α,， ,IL,-,6,， ,IL,-,1β, in the alcohol capsaicin group were significantly reduced （,P,<,0.05）， hepatic steatosis and intestinal mucosal pathological damage were reduced， mRNA expressions of intestinal mucosal inflammatory factors such as ,TNF,-,α,， ,IL,-,6, and ,MCP,-,1, in the alcohol capsaicin group mice were significantly down-regulated （,P,<,0.05）， while mRNA expressions of tight junction proteins including ,ZO,-,1,， ,occludin,， and ,claudin,-,1, were significantly up-regulated （,P,<,0.01）.,Conclusion,2,Capsaicin has a protective effect on the liver of mice with alcoholic liver disease， and its mechanism of action may be related to the intestinal mucosal barrier protection and intestinal leakage reduction.
关键词
辣椒素酒精性肝病肠黏膜屏障模型小鼠中药研究
Keywords
capsaicinalcoholic liver diseaseintestinal mucosal barriermodel micetraditional Chinese herbal medicine research
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