摘要
目的探讨硫化氢（H2S）对肠缺血／再灌注（I／R）损伤大鼠核转录因子-κB（NF-κB）及下游基因表达的影响。方法雄性Wistar大鼠24只，按随机数字表法分为假手术组（Sham组）、I／R组，I／R＋硫氢化钠（NaHS）组（NariS组），每组8只。NaHS组在再灌注前10rain静脉注射（静注）100μmol／kgNaHS后按1mg·kg^-1·h^-1持续静注直到再灌注2h，Sham组和I／R组静注等体积的生理盐水。用原位末端缺刻标记试验（TUNEL）观察回肠上皮细胞凋亡情况，并计算凋亡指数（AI），蛋白质免疫印迹试验（Western Blot）检测回肠组织NF-κB蛋白及NF-κB磷酸化（pNF-κB）水平，酶联免疫吸附试验（ELISA）测定血白细胞介素（IL-1、IL-6）、肿瘤坏死因子-α（TNF—α）水平，敏感硫电极法测定血H2S水平，动态浊度法测定血内毒素水平。采用直线相关分析法分析pNF—κB、TNF-α、IL-6、IL-1与H2S、AI、内毒素及内毒素、AI与H2S的相关性。结果Sham组未见凋亡细胞，I／R组可见较多的棕褐色凋亡回肠黏膜腺上皮细胞，AI明显增加[（39．62±4．50）％比（4．30±1．09）％]。NaHS组可见较稀疏的棕褐色凋亡回肠黏膜腺上皮细胞，AI较I／R组明显降低[（24．41±2．76）％比（39．62±4．50）％，P〈0．01]。与Sham组比较，I／R组NF—κB蛋白表达降低（灰度值：0．82±0．08比0．87±0．07），pNF—κB（灰度值：0．82±0．07比0．15±0．02）、TNF—α（ng／L：49．02±1．62比4．08±0．94）、IL-6（ng／L：437．08±12．43比227．97±12．92）、IL-1（96．12±12．35比47．97±9．68）、内毒素均明显升高（U／L：0．710±0．071比0，302±0．069），H，S明显降低（μmol／L：24．38±2．69比42，57±7．18）；与I／R组比较，NaHS组TNF—α（ng/39．27±1．54）、IL-6（ng／L：349．43±17．20）、IL-1（ng／L：67．38±14．57）、pNF—κB（0．78±0．09）、内毒素（U／L：0．558±
Objective To study the influence of hydrogen sulfide （HzS） on expressions of nuclear factor-κB （NF-κB） and downstream gene in rats with intestinal ischemia/reperfusion （I/R） injury. Methods Twenty-four male Wistar rats were randomly divided into sham operation group （Sham group）, I/R group, [/R and sodium hydrosulfide （NariS） group （n = 8 in each group）. In NariS group, 10 minutes prior to the onset of reperfusion, intravenous injection of 100 μmol/kg Naris was given to the rat, and afterwards 1 mg· kg^-1· h^-1 of Naris was persistently infused into the vein until reperfusion for 2 hours, whereas in Sham and I/R groups, equal volume of normal saline was intravenously infused to the rat. Ileal epithelial cell apoptosis was studied by TdT-mediated dUTP nick end labeling staining （TUNEL） and apoptosis index （AI） was calculated. The protein expression of ileal tissue NF-κB protein and the level of phosphorylated NF-κB （pNF-κB） were determined by Western Blot. The levels of serum interleukin （IL-1, IL-6）, tumor necrosis factor-α （TNF- α） were measured by enzyme-linked immunosorbent assay （ELISA）. The content of plasma H2S was tested by sensitive sulfide electrode. Endotoxin was detected by the dynamic turbidity method. The correlation between pNF-κB, TNF-α, IL-6, IL-1 and H2S, AI, endotoxin, and endotoxin, AI and H2S were analysised. Results No apoptotic ileal epithelial cells were seen in Sham group, while in I/R group, the brown apoptotic ileal epithelial cell was significantly increased, and AI was also obviously elevated [（39.62 ± 4.50）% vs. （4.30± 1.09）%]. In the NariS group,sparsely distributed brown apoptotic ileal nmcosal glandular epithelial ceils could be seen, and AI was significantly lower than that in I/R group [（24.41 ±2.76）% vs. （39.62 ±4.50）%, P 〈 0.01]. Compared with Sham group, the protein expression of NF-κB （gray value： 0.82±0.07 vs. 0.15±0.02）, pNF-κB （gray value： 0.82 ±0.07 vs. 0.15± 0.02）, TNF- α
出处
《中国中西医结合急救杂志》
CAS
北大核心
2016年第3期265-268,共4页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
国家自然科学基金青年科学基金项目（81201672）
浙江省义乌市引进人才科研项目（2012-R-04）
