摘要
目的探究类固醇受体辅活化子-3(SRC-3)对严重烧伤小鼠肠黏膜屏障功能损伤的影响。方法2022年1―4月将SPF级雌性SRC-3基因敲除(SRC-3-/-)小鼠作为对实验组(n=36),并以野生型(SRC-3+/+)小鼠作为对照组(n=36),两组小鼠均诱导建立背部30%体表总面积(TBSA)Ⅲ度烧伤模型。在烧伤后第1、3、5天时,荧光素异硫氰酸酯-葡聚糖(FITC-dextran)灌胃检测肠道通透性,酶联免疫吸附测定(ELISA)法检测血清白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平和血浆二胺氧化酶(DAO)活性以及内毒素(ET)水平,苏木精-伊红(HE)和阿尔新蓝-过碘酸-希夫(AB-PAS)染色评估肠黏膜损伤和杯状细胞黏液分泌情况,免疫组织化学(IHC)染色检测黏蛋白2(Muc2)的表达,蛋白质印迹法检测肠黏膜中Muc2、IL-6和TNF-α蛋白表达。结果在烧伤后第1、3、5天时,与对照组(SRC-3+/+小鼠)相比,实验组SRC-3-/-小鼠血清FITC-dextran浓度[(1156.21±107.65)μg/L比(685.14±79.36)μg/L、(1425.81±115.36)μg/L比(743.72±82.29)μg/L、(1613.27±120.94)μg/L比(824.35±85.44)μg/L]、血浆DAO活性和ET水平、肠黏膜损伤评分、PAS+杯状细胞数量、血清和肠黏膜中IL-6、TNF-α水平显著升高,AB+杯状细胞数量、肠黏膜中Muc2表达水平显著降低(P<0.05)。结论SRC-3缺失可以在严重烧伤后损害杯状细胞的分化成熟,减少肠黏液的合成与分泌,加重肠黏膜屏障功能障碍。
Objective To investigate the impact of steroid receptor coactivator-3(SRC-3)on the damage of intestinal mucosal barrier function in severely burned mice.Methods From January to April 2022,the SPF female SRC-3 knockout(SRC-3-/-)mice were regard⁃ed as the experimental group(n=36),and the wild-type(SRC-3+/+)mice were regarded as the control group(n=36),both groups of mice were induced to establish a 30%total body surface area(TBSA)third-degree burn model on the back.On the 1st,3rd,and 5th days af⁃ter burn,fluorescein isothiocyanate-dextran(FITC-dextran)was administered by gavage to detect intestinal permeability,the levels of serum interleukin-6(IL-6),tumor necrosis factor alpha(TNF-α),plasma diamine oxidase(DAO)activity,and the level of endotoxin(ET)were detected by enzyme-linked immunosorbent assay(ELISA),hematoxylin-eosin(HE)and alcian blue-periodic acid-Schiff(ABPAS)staining were used to evaluate intestinal mucosal injury and goblet cell mucus secretion,the expression of mucin 2(Muc2)was de⁃tected by immunohistochemical(IHC)staining,Western blotting was used to detect the protein expressions of Muc2,IL-6 and TNF-αin intestinal mucosa.Results On the 1st,3rd,and 5th days after burn,compared with the control group(SRC-3+/+mice),the serum FITCdextran concentration[(1156.21±107.65)μg/L vs.(685.14±79.36)μg/L,(1425.81±115.36)μg/L vs.(743.72±82.29)μg/L,(1613.27±120.94)μg/L vs.(824.35±85.44)μg/L],plasma DAO activity,level of ET,intestinal mucosal injury score,the number of PAS+goblet cells,the levels of IL-6 and TNF-αin serum and intestinal mucosa of SRC-3-/-mice in the experimental group were obviously increased,the number of AB+goblet cells and the expression level of Muc2 in intestinal mucosa were obviously decreased(P<0.05).Conclusion SRC-3 deletion can impair the differentiation and maturation of goblet cells after severe burn,reduce the synthesis and secretion of in⁃testinal mucus,and aggravate the dysfunction of intestinal mucosal barrier.
作者
杨宇
易庆军
李军
张永洪
文婷婷
YANG Yu;YI Qingjun;LI Jun;ZHANG Yonghong;WEN Tingting(Department of Anesthesiology,Mianyang Central Hospital,The Affiliated Mianyang Hospital,School of Medcine of UESTC,Mianyang,Sichuan 621000,China;Department of Pediatrics,Mianyang Central Hospital,The Affiliated Mianyang Hospital,School of Medcine of UESTC,Mianyang,Sichuan 621000,China)
出处
《安徽医药》
CAS
2024年第2期235-240,I0002,共7页
Anhui Medical and Pharmaceutical Journal
基金
创伤、烧伤与复合伤国家重点实验室开放基金资助(SKLKF202014)。
作者简介
通信作者:易庆军,女,副主任医师,研究方向为儿科学,Email:cgjus10@163.com。
