切换至 "中华医学电子期刊资源库"
高级检索
中华卫生应急电子杂志

中华卫生应急电子杂志 ›› 2023, Vol. 09 ›› Issue (02) : 90 -95. doi: 10.3877/cma.j.issn.2095-9133.2023.02.005

论著

艾司洛尔对脓毒症肠损伤的保护作用及对自噬蛋白AMPK表达水平的影响
李正达, 张艳兵, 刘茂霞, 李玉芳†(), 杨新静†()   
  1. 215100　江苏苏州，苏州大学附属第一医院重症医学科
    215100　江苏苏州，苏州大学附属第一医院麻醉科
  • 收稿日期:2023-03-02 出版日期:2023-04-18
  • 通信作者: 李玉芳, 杨新静
  • 基金资助:
    中华国际医学交流基金会心血管多学科整合思维研究基金项目(Z-2016-23-2001-46)

Protective effect of esmolol on sepsis-induced intestinal injury and its mechanism

Zhengda Li, Yanbin Zhang, Maoxia Liu, Xinjing Yang†()   

  1. Department of Intensive Care Unit, the First Affiliated Hospital of Soochow University, Suzhou 215100, China
    Department of Anesthesiology, the First Affiliated Hospital of Soochow University, Suzhou 215100, China
  • Received:2023-03-02 Published:2023-04-18
  • Corresponding author: Xinjing Yang
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

李正达, 张艳兵, 刘茂霞, 李玉芳, 杨新静. 艾司洛尔对脓毒症肠损伤的保护作用及对自噬蛋白AMPK表达水平的影响[J/OL]. 中华卫生应急电子杂志, 2023, 09(02): 90-95.

Zhengda Li, Yanbin Zhang, Maoxia Liu, Xinjing Yang. Protective effect of esmolol on sepsis-induced intestinal injury and its mechanism[J/OL]. Chinese Journal of Hygiene Rescue(Electronic Edition), 2023, 09(02): 90-95.

目的

探讨艾司洛尔(ES)对脓毒症大鼠肠损伤的保护作用及其具体作用机制。

方法

选择18只成年雄性SD大鼠，按随机数字表法分为假手术(Sham)组、脂多糖组(LPS)组和ES干预组(LPS+ES组)，每组6只。LPS组、LPS+ES组采用腹腔注射LPS(10 mg/kg)建立脓毒症肠损伤模型；Sham组注射等量等渗盐水。LPS+ES组持续经大鼠左颈内静脉泵入ES注射液(15 mg·kg-1·h-1)；Sham组、LPS组持续泵入等量等渗盐水。模型构建成功后12 h处死大鼠，并留取血标本及小肠组织。采用酶联免疫吸附法(ELISA)检测肠型脂肪酸结合蛋白(I-FABP)和二胺氧化酶(DAO)；苏木素-伊红(HE)染色观察小肠组织病理学变化；ELISA检测白介素-6(IL-6)和白介素-10(IL-10)水平；Western blotting检测Beclin-1、LC3-Ⅱ及p-AMPK的蛋白表达水平。

结果

与Sham组相比，LPS组光镜下可见小肠黏膜下层、肌层广泛空泡化变性，炎症细胞浸润，绒毛缩短，结构模糊；与LPS组相比，ES干预后黏膜下层、肌层广泛空泡化变性及炎症细胞浸润明显减轻，绒毛变长，病理损伤减轻。LPS组IL-6、IL-10、I-FABP及DAO较Sham组显著升高[两组分别为：(131.00±8.67)pg/mL比(106.40±6.70)pg/mL；(116.62±9.09)pg/mL比(103.23±7.38)pg/mL；(9.61±1.70)ng/mL比(3.91±0.70)ng/mL；(234.30±30.14)ng/mL比(37.49±12.11)ng/mL，P均<0.05]；ES干预后IL-6、I-FABP及DAO水平较LPS组明显降低[(117.50±9.00)pg/mL比(131.0±8.67)pg/mL；(5.34±1.10)pg/mL比(9.61±1.70)pg/mL；(147.80±17.07)ng/mL比(234.30±30.14)ng/mL，P均<0.05]，而IL-10水平较LPS组明显升高[(129.74±10.94)pg/mL比(116.62±9.09)pg/mL，P<0.05]。与Sham组相比，LPS组小肠组织Beclin-1、LC3-Ⅱ及p-AMPK的蛋白表达水平均明显降低(P<0.05)；与LPS组相比，ES干预后Beclin-1、LC3-Ⅱ及p-AMPK的蛋白表达量均明显增加(P均<0.05)。

结论

ES对脓毒症大鼠急性肠损伤有显著的保护作用，其机制可能是通过促进AMPK介导的自噬及降低炎症反应有关。

关键词: 脓毒症, 肠损伤, 脂多糖, 艾司洛尔, 自噬, AMPK
Objective

To investigate the protective effect and specific mechanism of esmolol(ES) on sepsis-induced intestinal injury in rats.

Methods

Eighteen adult male SD rats were randomLy divided into three groups according to the random number table method: Sham group(6 rats), lipopolysaccharide(LPS) group(6 rats), and LPS+ ES group(6 rats). The sepsis-induced acute intestinal injury rat model was reproduced by intraperitoneal injection of lipopolysaccharide (LPS)(10 mg/kg). The Sham group only received an equivalent amount of saline. In LPS+ ES group, esmolol dilution (15 mg·kg-1·h-1) was continuously pumped through left internal jugular vein; normal saline was continuously pumped into Sham group and LPS group. The rats were sacrificed when the model was successfully constructed for 12 h. The blood samples and the small intestinal tissues were collected. The diamine oxidase (DAO) and intestinal fatty acid-binding protein(I-FABP) were examined by enzyme-linked immunosorbent assay(ELISA). The intestinal tissue injury was analyzed by HE staining. The concentration of (interleukin-6, IL-6) and (interleukin-10, IL-10) were measured by ELISA. The expression levels of Beclin-1, LC3-Ⅱ, and p-AMPK was detected by Western blotting.

Results

Compared with Sham group, LPS group showed extensive vacuolization and inflammatory cell infiltration in submucosa and muscularis of small intestine, shortening of villi and blurring of structure under light microscope. After esmolol intervention, extensive vacuolization and inflammatory cell infiltration in submucosa and muscle layer were significantly reduced, villi became longer, and pathological damage was reduced. In LPS group, the levels of IL-6, IL-10, I-FABP and DAO were significantly higher than that of Sham group [(131.00±8.67)pg/mL vs (106.40±6.70)pg/mL; (116.62±9.09)pg/mL vs (103.23±7.38)pg/mL; (9.61±1.70)ng/mL vs (3.91±0.70)ng/mL; (234.30±30.14)ng/mL vs (37.49±12.11)ng/mL, respectively, all P<0.05]. The levels of IL-6, DAO, I-FABP in the LPS+ ES group were significantly decreased as compared with those in LPS group[(117.50±9.00)pg/mL vs 131.0±8.67)pg/mL; (5.34±1.10)pg/mL vs (9.61±1.70)pg/mL; (147.80±17.07)ng/mL vs (234.30±30.14)ng/mL, respectively, all P<0.05], but the expression of IL-10 was significantly increased in LPS+ ES group compared with that in LPS group [(129.74±10.94)pg/mL vs (116.62±9.09)pg/mL, P<0.05]. At the same time, compared with LPS group, the levels of Beclin-1, LC3-Ⅱ and p-AMPK was significantly decreased (all P<0.05), but the expression of Beclin-1, LC3-Ⅱ and p-AMPK protein was significantly higer in LPS+ ES group than those in LPS group(all P<0.05).

Conclusion

The protective effects of esmolol on sepsis-induced intestinal injury may be attributed to the up-regulation of AMPK-mediated autophagy and reduction of inflammatory response.

Key words: Sepsis, Intestinal injury, Lipopolysaccharide, Esmolol, Autophagy, AMPK
图1 光镜下观察各组大鼠肠组织病理学变化(HE染色，×100)注：Sham为假手术组，LPS为脂多糖组，LPS+ES为脂多糖+艾司洛尔组
图2 光镜下观察各组大鼠肠绒毛变化注：Sham为假手术组，LPS为脂多糖，LPS+ES为脂多糖+艾司洛尔；ΔP<0.05 vs Sham group；*P<0.05 vs LPS group，n＝6
图3 3组肠组织IL-6和IL-10表达水平的比较(n＝6)注：Sham为假手术组，LPS为脂多糖，LPS+ES为脂多糖+艾司洛尔；ΔP<0.05 vs Sham group；*P<0.05 vs LPS group，n＝6
图4 3组肠组织I-FABP及DAO的表达水平的比较(n＝6)注：Sham为假手术组，LPS为脂多糖，LPS+ES为脂多糖+艾司洛尔；ΔP<0.05 vs Sham group；*P<0.05 vs LPS group，n＝6
图5 3组肠组织Beclin-1、LC3-Ⅱ及p-AMPK的蛋白表达水平的比较(n＝6)注：Sham为假手术组，LPS为脂多糖，LPS+ES为脂多糖+艾司洛尔；ΔP<0.05 vs Sham group；*P<0.05 vs LPS group，n＝6
1
Evans L，Rhodes A，Alhazzani W，et al.Surviving sepsis campaign：international guidelines for management of sepsis and septic shock 2021[J]．Intensive Care Med，2021，47(11)：1181-1247.
2
Sun S，Duan Z，Wang X，et al.Neutrophil extracellular traps impair intestinal barrier functions in sepsis by regulating TLR9-mediated endoplasmic reticulum stress pathway[J]．Cell Death Dis，2021，12(6)：606.
3
Padar M，Starkopf J，Uusvel G，et al.Gastrointestinal failure affects outcome of intensive care[J]．J Crit Care，2019(52)：103-108.
4
Morelli A，Singer M，Ranieri VM，et al.Heart rate reduction with esmolol is associated with improved arterial elastance in patients with septic shock：a prospective observational study[J]．Intensive Care Med，2016，42(10)：1528-1534.
5
Mori K，Morisaki H，Yajima S，et al.Beta-1 blocker improves survival of septic rats through preservation of gut barrier function[J]．Intensive Care Med，2011，37(11)：1849-1856.
6
Otani S，Coopersmith CM.Gut integrity in critical illness[J]．J Intensive Care，2019(7)：17.
7
Durand M，Hagimont E，Louis H，et al.The β1-Adrenergic receptor contributes to sepsis-induced immunosuppression through modulation of regulatory t-cell inhibitory function[J]．Crit Care Med，2022，50(9)：e707-e718.
8
Suzuki T，Inoue K，Igarashi T，et al.Beta-blocker therapy preserves normal splenic T-lymphocyte numbers reduced in proportion to sepsis severity in a sepsis model[J]．Crit Care Res Pract，2019(2019)：8157482.
9
Levy B，Fritz C，Piona C，et al.Hemodynamic and anti-inflammatory effects of early esmolol use in hyperkinetic septic shock：a pilot study[J]．Crit Care，2021，25(1)：21.
10
Tan S，Zhou F，Zhang Z，et al.Beta-1 blocker reduces inflammation and preserves intestinal barrier function after open abdominal surgery[J]．Surgery，2021，169(4)：885-893.
11
张璐瑶，聂垚，柯路，等．艾司洛尔对脓毒症大鼠肠黏膜屏障的影响[J]．肠外与肠内营养，2014，21(5)：305-308.
12
Seethaler B，Basrai M，Neyrinck AM，et al.Biomarkers for assessment of intestinal permeability in clinical practice[J]．Am J Physiol Gastrointest Liver Physiol，2021，321(1)：G11-G17.
13
张剑，徐俊，洪丽华，等．肠型脂肪酸结合蛋白，二胺氧化酶在绞窄性肠梗阻大鼠体内与肠管损伤的相关性[J]．中华内分泌外科杂志，2021，15(6)：598-602.
14
Xu Z，Li D，Qu W，et al.Card9 protects sepsis by regulating Ripk2-mediated activation of NLRP3 inflammasome in macrophages[J]．Cell Death Dis，2022，13(5)：502.
15
刘委宏，郝浩，刘阳，等．半夏泻心汤通过调控HO-1的表达改善脓毒症大鼠肠道损伤的机制研究[J]．中国中医急症，2021，30(6)：945-949.
16
张红涛，于泳浩，马小叶，等．氢气吸入对严重脓毒症小鼠血清炎性因子和肠损伤的影响[J]．中华危重病急救医学，2015，27(6)：498-503.
17
Morhardt TL，Hayashi A，Ochi T，et al.IL-10 produced by macrophages regulates epithelial integrity in the small intestine[J]．Sci Rep，2019，9(1)：1223.
18
Foerster EG，Mukherjee T，Cabral-Fernandes L，et al.How autophagy controls the intestinal epithelial barrier[J]．Autophagy，2022，18(1)：86-103.
19
Zhou Y，Wang Z，Huang Y，et al.Membrane dynamics of ATG4B and LC3 in autophagosome formation[J]．J Mol Cell Biol，2022，13(12)：853-863.
20
李小静，李志锋，李显平，等．丹参酮ⅡA体外促进黑素瘤A375细胞自噬及信号通路的实验研究[J]．中华皮肤科杂志，2017，50(1)：29-32
21
Wang JF，Mei ZG，Fu Y，et al.Puerarin protects rat brain against ischemia/reperfusion injury by suppressing autophagy via the AMPK-mTOR-ULK1 signaling pathway[J]．Neural Regen Res，2018，13(6)：989-998.
22
Wan SX，Shi B，Lou XL，et al.Ghrelin protects small intestinal epithelium against sepsis-induced injury by enhancing the autophagy of intestinal epithelial cells[J]．Biomed Pharmacother，2016(83)：1315-1320.
23
Yang XJ，Qian JX，Wei Y，et al.Tanshinone IIA sodium sulfonate attenuates LPS-induced intestinal injury in mice[J]．Gastroenterol Res Pract，2018(2018)：9867150.
24
Li Q，Li L，Fei X，et al.Inhibition of autophagy with 3-methyladenine is protective in a lethal model of murine endotoxemia and polymicrobial sepsis[J]．Innate Immun，2018，24(4)：231-239.
[1] 吴杰, 周志强, 符菁, 李喜功, 张钦. 吸入性氢气对大鼠脊髓损伤后自噬及神经功能的影响[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(05): 363-371.
[2] 庄燕, 戴林峰, 张海东, 陈秋华, 聂清芳. 脓毒症患者早期生存影响因素及Cox 风险预测模型构建[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(05): 372-378.
[3] 杨瑾, 刘雪克, 张媛媛, 金钧, 韦瑶. 肠道微生物来源石胆酸对脓毒症相关肝损伤的保护作用[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(04): 265-274.
[4] 张霞, 张瑞, 郑志波, 张勤. 紫草素调控乳酸化修饰和线粒体功能改善脓毒症心肌病小鼠的预后[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(04): 275-284.
[5] 张婧琦, 江洋, 孙佳璐, 唐兴喆, 赵宇飞, 崔颖, 李信响, 戴景月, 傅琳, 彭新桂. 基于肾周CT特征结合血清肌酐水平探讨脓毒症伴急性肾损伤的早期识别[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(04): 285-292.
[6] 李振翮, 魏长青, 甄国栋, 李振富. 脓毒症并发急性呼吸窘迫综合征患者血清S1P、Wnt5a变化及其临床意义[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(04): 293-300.
[7] 李争光, 宰爽嘉, 吴火峰, 孙华, 张永博, 陈浏阳, 戴睿, 张亮. 昼夜节律相关因子在椎间盘退行性变发病机制中作用的研究进展[J/OL]. 中华损伤与修复杂志(电子版), 2024, 19(05): 457-461.
[8] 廖泽楷, 梁爱琳, 龚启梅. 根尖周病中程序性细胞死亡的研究进展[J/OL]. 中华口腔医学研究杂志(电子版), 2024, 18(03): 150-155.
[9] 成人脓毒症患者β-内酰胺类抗生素延长输注专家共识编写组. 成人脓毒症患者β-内酰胺类抗生素延长输注专家共识[J/OL]. 中华重症医学电子杂志, 2024, 10(04): 313-324.
[10] 陈曦, 吴宗盛, 郑明珠, 邱海波. 胸腺萎缩在脓毒症免疫紊乱中的研究进展[J/OL]. 中华重症医学电子杂志, 2024, 10(04): 379-383.
[11] 杨翔, 郭兰骐, 谢剑锋, 邱海波. 转录组学在脓毒症诊疗中的临床研究进展[J/OL]. 中华重症医学电子杂志, 2024, 10(04): 384-388.
[12] 史清泉, 苗彬, 王烁, 陶琳, 沈晨. miR-181a-5p 靶向ATG5 抑制雨蛙素诱导的大鼠胰腺腺泡细胞AR42J自噬的机制研究[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(06): 524-530.
[13] 陈惠英, 邱敏珊, 邵汉权. 脓毒症诱发肠黏膜屏障功能损伤的风险因素模型构建与应用效果[J/OL]. 中华消化病与影像杂志(电子版), 2024, 14(05): 448-452.
[14] 傅新露, 李之岳, 卢丹. 妊娠合并结肠癌穿孔致脓毒症休克一例并文献复习[J/OL]. 中华产科急救电子杂志, 2024, 13(04): 227-231.
[15] 刘霖, 张文欢, 宋雅茹, 姜云璐. Apelin-13 在阿尔茨海默病中的神经保护作用机制研究进展[J/OL]. 中华诊断学电子杂志, 2024, 12(04): 276-280.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号