摘要
目的观察钠氢交换蛋白1（NHE1）抑制剂对烧伤脓毒症大鼠肠道损伤的作用，并初步探讨其可能机制。方法取SD大鼠90只，按照随机数字表法分为对照组、单纯脓毒症组和NHE1抑制剂组，每组30只。单纯脓毒症组和NHE1抑制剂组大鼠背部建立20％TBSAⅢ度烫伤（下称烧伤）模型后，背部创面中心注射2×10^5CFU／mL的铜绿假单胞菌ATCC27853菌液50μL。烧伤脓毒症模型建立成功后，NHE1抑制剂组大鼠迅速腹腔注射0．1mmol／LNHE1抑制剂卡立泊来德0．4mg／kg，单纯脓毒症组大鼠注射同体积的生理盐水。对照组大鼠除不制作烧伤创面和接种细菌外，其余操作与单纯脓毒症组大鼠相同。烧伤脓毒症大鼠在伤后12h剖腹，于回肠远端注射0．1mol／L异硫氰酸荧光素（FITC）-葡聚糖200mL，30min后左心室采血，切取回肠末端组织。荧光分光光度计检测大鼠血清FITC-葡聚糖含量（样本数为10），HE染色观察肠道组织形态（样本数为10），ELISA法检测血清和肠道组织IL-6和TNF—α含量（样本数均为20），比色法检测血清和肠道组织髓过氧化物酶（MPO）活性（样本数均为20），蛋白质印迹法检测肠道组织NF-κB-p65蛋白表达及MAPK信号通路相关蛋白p38MAPK、细胞外信号调节激酶1／2（ERK1／2）以及c—Jun氨基末端激酶1／2（JNK1／2）磷酸化水平（样本数为4）。对照组大鼠于相同时相点同前采集标本进行相关检测。对数据行单因素方差分析、SNK检验。结果（1）单纯脓毒症组大鼠血清FITC-葡聚糖含量较对照组明显升高（P〈0．01），NHE1抑制剂组大鼠血清FITC-葡聚糖含量则较单纯脓毒症组显著降低（P〈0．01）。与对照组比较，单纯脓毒症组大鼠肠黏膜可见大量炎性细胞浸润、黏膜溃疡以及黏膜坏死；NHE1抑制剂组大鼠肠道损伤情况较单纯脓毒症组有显著好转。（2）单纯脓毒症组大鼠血清IL-6、TNF-α、MPO
Objective To observe the effects of Na ±/H ± exchanger 1 （ NHE1 ） inhibitor on intesti- nal injury of rats with burn sepsis, and to explore the possible mechanism preliminarily. Methods Ninety SD rats were divided into control group, pure sepsis group, and NHE1 inhibitor group according to the random number table, with 30 rats in each group. Full-thickness scald （hereinafter referred to as burn） model with 20% total body surface area were reproduced on the back of rats in pure sepsis and NHE1 inhibitor groups, and then 50 μL liquid of Pseudomonas aeruginosa ATCC 27853 （2 × 10^5 colony forming unit/mL） were injected into the center of wounds on the back. Rats in NHE1 inhibitor group were intraperitoneally injected with 0.1 mmol/L NHEI inhibitor cariporide （0.4 mg/kg） rapidly after the successful establishment of burn sepsis model, while rats in pure sepsis group were injected with the same volume of normal saline. Except for not being made burn wounds nor receiving bacterination, rats in control group were treated the same as those in pure sepsis group. Rats with burn sepsis in each group were laparotomized and injected with 200 mL fluorescein isothiocyanate （FITC）-dextran in the concentration of 0.1 mol/L in terminal ileum at 12 hours post injury, and their left ventricular blood and terminal ileum were collected 30 minutes later. The serum content of FITC-dextran was detected with fluorescence spectrophotometer （ n = 10） ; the morphology of intestinal tissue was observed with HE staining （ n = 10） ; the content of interleukin-6 （IL-6） and tumor necrosis factor-alpha （TNF-α） in serum and intestinal tissue was determined with enzyme-linked immunosor- bent assay （ n = 20） ; the activity of myeloperoxidase （MPO） in serum and intestinal tissue was detected with colorimetric method （ n = 20） ; the protein expression of nuclear factor-kappa B-p65 （NF-KB-p65） and phosphorylation levels of mitogen-activated protein kinase （MAPK） signal pathway related proteins
作者
李文鹏
赵高炀
杨薛康
Li Wenpeng Zhao Gaoyang Yang Xuekang(Student Brigade, the Fourth Military Medical Universi- ty, Xi＇an 710032, China)
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2017年第6期349-354,共6页
Chinese Journal of Burns
基金
国家自然科学基金（81201463）
