摘要
目的观察重复应用镇痛剂量艾司氯胺酮(Esk)对严重烧伤小鼠肠损伤的影响。方法成年雄性无特定病原体(SPF)级C57BL/6J小鼠(江苏集萃药康生物有限公司)采用随机数字表法分为假烧伤组(Sham组)、烧伤模型组(Mod组)、假烧伤+艾司氯胺酮组(Sham+Esk组)和烧伤+艾司氯胺酮组(Mod+Esk组),制备40%全身体表面积深Ⅱ度烧伤模型。Sham+Esk组及Mod+Esk组给予腹腔注射艾司氯胺酮0.01 mg/g,观察建模前后多个时点的疼痛应激评分、糖水偏好指数及8 d生存率(n=10)。造模后22 h以异硫氰酸荧光素标记的右旋糖酐(FITC-dextran)灌胃(n=4)。造模后24 h处死小鼠收集血清比较FITC-dextran浓度,观察小肠Chiu’s损伤评分、糖原(+)细胞数和原位缺口末端标记法(TUNEL)凋亡指数,测定小肠组织急性炎症指标肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β含量和小肠紧密连接蛋白(Claudin 7和ZO-1)表达水平(n=6)。同时点指标的多组间比较通过单因素方差分析或非参数检验,两组间比较采用t检验或Mann-Whitney检验。结果Mod组小鼠小肠黏膜凋亡指数高于Sham组[(28.3±7.2)%比(3.1±2.3)%,t=12.82,P<0.01],血清FITC-dextran浓度高于Sham组[(0.66±0.01)mg/ml比(0.60±0.01)mg/ml,t=11.63,P<0.01];肠壁绒毛平均糖原(+)细胞数低于Sham组[(7.3±3.0)个比(29.1±5.5)个,t=5.78,P<0.01],ZO-1蛋白的相对强度低于Sham组(0.3±0.2比1.2±0.1,t=5.53,P<0.01)。给予Esk治疗后烧伤小鼠的小肠黏膜损伤减轻:Mod+Esk组肠壁凋亡指数低于Mod组[(14.5±4.6)%比(28.3±7.2)%,t=12.82,P<0.01],血清FITC-dextran浓度低于Mod组[(0.63±0.01)mg/ml比(0.66±0.01)mg/ml,t=5.768,P<0.01];糖原(+)细胞数高于Mod组[(14.1±3.5)个比(7.3±3.0)个,t=5.78,P<0.01],Claudin 7蛋白相对强度也高于Mod组(1.3±0.2比0.6±0.4,t=3.07,P<0.05)。结论镇痛剂量艾司氯胺酮可减轻严重烧伤小鼠肠屏障损伤,机制可能与促进肠黏液分泌和改善肠道炎症有关。
Objective To evaluate the effect of repeated analgesic doses of S-ketamine(Esk)on intestinal injury in severely burned mice.MethodsAdult male specific pathogen free(SPF)C57BL/6J mice were randomly divided into four groups:sham burn group(Sham group),severe burn model group(Mod group),sham burn+Esk group(Sham+Esk group),and severe burn+Esk group(Mod+Esk group).A 40%total body surface area deep second degree burn model was prepared.The Sham+Esk and Mod+Esk groups received intraperitoneal injections of Esk at a dose of 10 mg/kg.Pain and distress scale,surcrose preference(SP)test,and 8-day survival rate were observed at multiple time points before and after modeling(n=10).Mice were gavaged with fluorescein isothiocyanate-labeled dextran(FITC-dextran)at 22 h after modeling(n=4).At 24 h post-modeling,mice were euthanized,and serum was collected to compare FITC-dextran concentrations.Meanwhile,intestinal Chiu’s injury scores,glycogen(+)cell counts,terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)apoptosis index,intestine tumor necrosis factor(TNF)-αand interleukin(IL)-1βlevels,and intestinal tight junction proteins(Claudin 7 and ZO-1)levels were measured(n=6).Multiple group comparisons at the same time points were performed using one-way analysis of variance or non-parametric tests,and comparisons between two groups were conducted using t-tests or Mann-Whitney tests.ResultsThe apoptosis index of the small intestine mucosa in Mod group was higher than that in Sham group[(28.3±7.2)%vs.(3.1±2.3)%,t=12.82,P<0.01],and the serum FITC-dextran concentration was higher than that in Sham group[(0.66±0.01)vs.(0.60±0.01)mg/ml,t=11.63,P<0.01];The average number of glycogen(+)cells in a single intestinal villus was less in the Mod group(7.3±3.0 vs.29.1±5.5,t=5.78,P<0.01),and the relative intensity of ZO-1 protein was lower than in the Sham group(0.3±0.2 vs.1.2±0.1,t=5.53,P<0.01).After Esk treatment,the intestinal apoptosis index in the Mod+Esk group decreased compared to the Mod group[(14.5±4.6)%vs.(28.3±7.2)%,t=12.82,P<0.01],and the serum FITC-dextran concentration was lower than that in the Mod group[(0.63±0.01)vs.(0.66±0.01)mg/ml,t=5.768,P<0.01];The number of glycogen(+)cells and the relative strength of Claudin 7 protein were higher than those in the Mod group(14.1±3.5 vs.7.3±3.0,t=5.78,P<0.01;1.3±0.2 vs.0.6±0.4,t=3.07,P<0.05).ConclusionAnalgesic dose of S-ketamine can alleviate severe burn-induced intestinal barrier injury in mice.The mechanism may be associated with promoting intestinal mucus secretion and improving intestinal inflammation.
作者
贺秋兰
刘琦
陈彩云
温鲁平
周飞
黄文起
He Qiulan;Liu Qi;Chen Caiyun;Wen Luping;Zhou Fei;Huang Wenqi(Department of Anesthesiology,First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China;Department of Anorectal Section,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221000,China;Department of Burn Surgery,First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China)
出处
《中华实验外科杂志》
CAS
2024年第3期472-476,共5页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金青年项目(82004368)。
关键词
艾司氯胺酮
烧伤
肠损伤
肠黏膜屏障
S-ketamine
Burns
Intestinal injury
Intestinal barriers
作者简介
通信作者:黄文起,Email:huangwq@mail.sysu.edu.cn。
