慢性束缚应激对小鼠小肠屏障的损伤研究  

Studies on the Damage of the Small Intestinal Barrier in Mice by Chronic Restraint Stress

作　　者：孙忠鑫 柴露露 刘秋宏 张尘龙 刘舜星 李慧 林如涛[1] SUN Zhongxin;CAI Lulu;LIU Qiuhong;ZHANG Chenlong;LIU Shunxing;LI Hui;LIN Rutao(College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵阳550025

出　　处：《西南大学学报（自然科学版）》2024年第2期52-61,共10页Journal of Southwest University(Natural Science Edition)

基　　金：贵州省科学基金项目(黔科合基础-ZK[2023]一般107);贵州大学博士基金项目(贵大人基合字(2020)号66号);国家自然科学基金青年项目(32102630)。

摘　　要：肠屏障由机械屏障、免疫屏障、化学屏障和生物屏障共同组成,在维持肠腔内环境稳态和肠上皮结构完整性等方面发挥重要作用.首先建构小鼠慢性束缚应激模型并进行血浆激素水平检测和小鼠旷场实验,验证模型是否建构成功.随后通过常规组织学染色、免疫组化、 TUNEL免疫荧光、 ELISA等方法研究慢性束缚应激对小鼠小肠的损伤作用.结果显示,慢性束缚应激显著提高小鼠血浆NE和CORT水平(p<0.05),降低小鼠小肠绒毛高度(V,p<0.05)、增加隐窝深度(C,p<0.05),降低V/C比值(p<0.05)及紧密连接蛋白ZO-1表达量(p<0.01);显著减少小肠杯状细胞和内皮单核淋巴细胞数量(p<0.01),提示慢性束缚应激造成了小鼠肠道损伤和免疫功能抑制.进一步研究发现,慢性应激小鼠肠道内PCNA阳性表达显著减少(p<0.01)、凋亡细胞显著增多(p<0.01);血浆中TNF-α,IL-1β,IL-18含量显著升高(p<0.01),而IL-10显著降低(p<0.01).血浆氧化-抗氧化指标结果显示,慢性束缚应激显著增加小鼠血浆MDA含量(p<0.05),降低T-AOC,SOD,CAT,GSH-Px(p<0.05)活性,提示慢性束缚应激诱导机体氧化应激.该研究表明慢性束缚应激通过引发机体氧化应激,造成肠上皮细胞更新抑制和肠道屏障结构损伤,加重机体炎症反应.The gut barrier consists of mechanical,immune,chemical and biological barriers,which plays an important role in maintaining the homeostasis of the intestinal lumen and the integrity of the intestina epithelium.Firstly,hormone level tests and open field test were used to verify whether the mouse chronic restraint stress model was constructed successfully.Subsequently,conventional histological staining,immunohistochemistry,TUNEL immunofluorescence and ELISA were used to investigated the effects of chronic restraint stress on mice gut barrier.Our results showed that compared with the control group,chronic restraint stress significantly increased plasma NE and CORT levels(p<0.05),decreased the height of small intestinal villi(V,p<0.05),increased the depth of crypts(C,p<0.05),and decreased the V/C ratio(p<0.05),the positive expression of the tight junction protein ZO-1(p<0.01),the number of small intestinal goblet cell and endothelial mononuclear lymphocytes(p<0.01),suggesting that chronic restraint stress caused intestinal damage and immune suppression in mice.Further studies revealed that chronic stress significantly decreased PCNA positive expression(p<0.01),while increased apoptotic cells numbers(p<0.01)in the small intestine.The plasma levels of TNF-α,IL-1β,and IL-18 were significantly increased(p<0.01),while IL-10 was significantly decreased(p<0.01)than that of control group.In addition,the results of plasma oxidation-antioxidant indexes showed that chronic restraint stress significantly increased MDA content(p<0.05)and decreased T-AOC,SOD,CAT and GSH-Px levels(p<0.05)in mice.This study demonstrated that chronic restraint stress can inhibit cell proliferation and damage the intestinal mucosal barrier function,aggravate inflammatory reactions by inducing oxidative stress in mice.

关 键 词：小鼠 慢性束缚应激 机械屏障 炎症反应 

分 类 号：S865.1[农业科学—野生动物驯养]