胃饥饿素抑制脓毒症大鼠的促炎反应并预防认知障碍。

Ghrelin inhibits proinflammatory responses and prevents cognitive impairment in septic rats.

机构信息

1Department of Anesthesiology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China. 2Department of Anesthesiology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. 3Shanghai Key Laboratory of Brain Functional Genomics, The Key Laboratory of Ministry of Education of China, East China Normal University, Shanghai, China. 4Department of Anesthesia and Perioperative Care, University of California, San Francisco, CA.

出版信息

Crit Care Med. 2015 May;43(5):e143-50. doi: 10.1097/CCM.0000000000000930.

DOI:10.1097/CCM.0000000000000930

PMID:25756415

Abstract

OBJECTIVES

A novel stomach-derived peptide, ghrelin, is down-regulated in sepsis and its IV administration decreases proinflammatory cytokines and mitigates organ injury. In this study, we wanted to investigate the effects of ghrelin on proinflammatory responses and cognitive impairment in septic rats.

DESIGN

Prospective, randomized, controlled experiment.

SETTING

Animal basic science laboratory.

SUBJECTS

Sprague-Dawley rats, weighing 250-300 g.

INTERVENTIONS

Sepsis was induced by cecal ligation and puncture. Animals were randomly divided into four groups: sham, sham + ghrelin, cecal ligation and puncture, and cecal ligation and puncture + ghrelin. Saline was given subcutaneously (30 mL/kg) at 4 and 16 hours after surgery for all rats. Septic rats were treated with ceftriaxone (30 mg/kg) and clindamycin (25 mg/kg) subcutaneously at 4 and 16 hours after surgery. Ghrelin (80 μg/kg) was administrated intraperitoneally 4 and 16 hours after surgery in sham + ghrelin group and cecal ligation and puncture + ghrelin group.

MEASUREMENTS AND MAIN RESULTS

The levels of proinflammatory cytokines in hippocampus were measured by enzyme-linked immunosorbent assay, and cleaved caspase-3 was detected by Western blot 24 hours after surgery. Neuronal apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick-end labeling staining 48 hours after surgery. Additional animals were monitored to record survival and body weight changes for 10 days after surgery. Survival animals underwent behavioral tasks 10 days after surgery: open-field, novel object recognition, and continuous multiple-trial step-down inhibitory avoidance task. Ghrelin significantly decreased the levels of proinflammatory cytokines and inhibited the activation of caspase-3 in the hippocampus after cecal ligation and puncture. The density of terminal deoxynucleotidyl transferase dUTP nick-end labeling-positive apoptotic neurons was significantly lowered by ghrelin. In addition, ghrelin improved the survival rates after cecal ligation and puncture. There were no differences in the distance and move time between groups in open-field task. However, the survivors after cecal ligation and puncture were unable to recognize the novel object and required more training trials to reach the acquisition criterion. All these long-term impairments were prevented by ghrelin.

CONCLUSIONS

Ghrelin inhibited proinflammatory responses, improved the survival rate, and prevented cognitive impairment in septic rats.

摘要

目的

一种新型的胃来源肽——ghrelin 在脓毒症中下调，其静脉给药可减少促炎细胞因子并减轻器官损伤。本研究旨在探讨 ghrelin 对脓毒症大鼠促炎反应和认知障碍的影响。

设计

前瞻性、随机、对照实验。

地点

动物基础科学实验室。

对象

体重 250-300 克的 Sprague-Dawley 大鼠。

干预

盲肠结扎穿孔术诱导脓毒症。动物随机分为四组：假手术、假手术+ghrelin、盲肠结扎穿孔术和盲肠结扎穿孔术+ghrelin。所有大鼠均于术后 4 小时和 16 小时皮下给予生理盐水（30ml/kg）。术后 4 小时和 16 小时，脓毒症大鼠皮下给予头孢曲松（30mg/kg）和克林霉素（25mg/kg）。假手术+ghrelin 组和盲肠结扎穿孔术+ghrelin 组于术后 4 小时和 16 小时腹腔内给予 ghrelin（80μg/kg）。

测量和主要结果

术后 24 小时通过酶联免疫吸附试验测量海马内促炎细胞因子水平，通过 Western blot 检测裂解的 caspase-3。术后 48 小时通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记染色法检测神经元凋亡。另外一些动物被监测以记录术后 10 天的存活和体重变化。术后 10 天，存活动物进行行为任务：旷场、新物体识别和连续多试抑制性避退任务。Ghrelin 显著降低盲肠结扎穿孔术后海马内促炎细胞因子水平，并抑制 caspase-3 的激活。Ghrelin 显著降低末端脱氧核苷酸转移酶 dUTP 缺口末端标记阳性凋亡神经元的密度。此外，ghrelin 提高了盲肠结扎穿孔术后的生存率。旷场任务中各组之间的距离和移动时间没有差异。然而，盲肠结扎穿孔术后的幸存者无法识别新物体，需要更多的训练试验才能达到获得标准。ghrelin 可预防所有这些长期损害。