摘要
目的探讨沉默信息调节因子1(SIRT1)在脂多糖(lipopolysaccharide,LPS)诱导的坏死性小肠结肠炎(NEC)小鼠的保护作用。方法 3-4日龄C57BL/6小鼠60只被随机分为三组。对照组大鼠与母鼠同笼,鼠乳喂养,不做任何处理; NEC模型组、SIRT1激动剂干预组大鼠出生24 h与母鼠分开,置于保育箱中,采用鼠乳代用品人工饲养,采用低温缺氧诱导NEC模型。模型制备结束后空腹12 h,处死小鼠采集十二指肠下端至回盲部肠管,观察肠管的弹性、色泽及肠腔有无积气、出血、坏死等病变; HE染色观察肠组织病理变化; ELISA测定肠组织IL-6、IL-10、TNF-α水平; qRT-PCR法检测IL-6、IL-10、TNF-αmRNA表达水平; Western blot检测肠组织SIRT1蛋白表达水平。结果模型组小鼠均出现不同程度的腹胀、排黄绿色稀便、嗜睡、胃滞留、呼吸急促等症状,SIRT1激动剂干预组小鼠各症状较模型组减轻,对照组小鼠无异常情况,模型组死亡7只,干预组无死亡。肠组织病理切片显示,对照组肠道上皮及肠绒毛完整,无血管扩张,模型组黏膜下层出血明显、水肿,黏膜下层和固有层分离,肠绒毛结构不完整,部分脱落、坏死,干预组较模型组损伤较轻。Western blot检测显示,与对照组比较,模型组、干预组小鼠肠组织SIRT1表达显著降低(P <0. 05),与模型组比较,SIRT1激动剂干预组SIRT1水平显著升高(P <0. 05)。ELISA检测显示,与对照组比较,模型组肠组织中IL-6、TNF-α水平显著升高,IL-10水平显著降低(P <0. 05);与模型组比较,干预组肠组织中IL-6、TNF-α、IL-1β水平显著降低,IL-10水平显著升高(P <0. 05)。qRT-PCR验证结果显示,模型组肠组织中IL-6、TNF-αmRNA水平较对照组显著升高,IL-10 mRNA水平显著降低(P <0. 05);与模型组比较,干预组肠组织中IL-6、TNF-αmRNA水平显著降低,IL-10 mRNA水平显著升高(P <0. 05)。结论 SIRT1在LPS诱导的坏死性小肠结肠炎小鼠肠内表达水平降低,SIRT1激�
Objective To investigate the protective effect of SIRT1 on lipopolysaccharide( LPS)-induced necrotizing enterocolitis(NEC) in mice. Methods Sixty C57 BL/6 mice aged 3-4 days were randomly divided into three groups. The mice in NEC model group and SIRT1 agonist group were separated from their mothers at 24 h after birth,and then placed in incubator and artificially reared with rat milk substitutes. At the same time,NEC model was induced by hypoxia-induced hypothermia. The mice in control group were fed in the same cage with their mothers without any treatment. Fasting for 12 h after model preparation,the mice were sacrificed to collect the intestinal tube from the lower duodenum to ileocecum. The elasticity and color of ileocecum,and pneumatosis,hemorrhage,necrosis changes in intestinal cavity were observed. The pathological changes of intestinal tissue were observed by HE staining. The levels of IL-6,IL-10 and TNF-α in intestinal tissue were measured by ELISA. The expression levels of IL-6,IL-10,and TNF-α mRNA were detected by qRT-PCR. SIRT1 protein expression level in intestinal tissue was detected by Western blot. Results The mice in model group showed abdominal distension,yellowish-green loose stool,lethargy,gastric retention,shortness of breath and other symptoms in varying degrees. The symptoms were lighter in SIRT1 agonist group than in model group,and there was no abnormality in control group. Seven mice died in model group and no one in SIRT1 agonist group. The pathological sections of intestinal tissues showed that intestinal epithelium and villi were intact in control group without vasodilatation,however,submucosal hemorrhage and edema were obvious in model group,in which the submucosal and lamina propria were separated and intestinal villus structure was incomplete with some of them being exfoliated and necrotic. The injury in SIRT1 agonist group was lighter than that in model group.Western blot analysis showed that the expression of SIRT1 in intestinal tissue were significantly lower in model group
作者
张岚
白铂亮
王莉
高琼
席朝霞
桂艳红
王惠萍
ZHANG Lan;BAI Boliang;WANG Li;GAO Qiong;XI Zhaoxia;GUI Yanhong;WANG Huiping(Department of Pediatrics, Second Affiliated Hospital of Xi’ an Jiaotong University, Xi’ an 710004, China)
出处
《山西医科大学学报》
CAS
2019年第5期570-575,共6页
Journal of Shanxi Medical University
基金
陕西省科学技术厅重点研发项目(2018SF-083)
