天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (9): 924-927.doi: 10.11958/20190382

• 实验研究 • 上一篇    下一篇

程序性坏死特异性抑制剂-1对呼吸机相关性肺损伤的保护作用

杜学柯，荆忍，张韵希，葛万运   

  1. 基金项目：广西高校中青年教师基础能力提升项目（2018KY0113）；广西自然科学基金资助（联合资助培育项目）（2018GXNSFAA138007）； 广西医疗卫生适宜技术开发与推广应用项目（S2018104） 作者单位：广西医科大学附属肿瘤医院麻醉科（邮编530021） 作者简介：杜学柯（1981），男，博士，副主任医师，主要从事围手术期肺损伤和肺功能保护研究
  • 收稿日期:2019-02-13 修回日期:2019-06-23 出版日期:2019-09-15 发布日期:2019-09-18
  • 通讯作者: 杜学柯 E-mail:46699644@qq.com
  • 基金资助:
    广西自然科学基金资助（联合资助培育项目）;广西医疗卫生适宜技术开发与推广应用项目;广西高校中青年教师基础能力提升项目

Protective effect of necrostatin-1 on the ventilator-induced lung injury in rats

DU Xue-ke, JING Ren, ZHANG Yun-xi, GE Wan-yun   

  1. Department of Anesthesiology, the Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China
  • Received:2019-02-13 Revised:2019-06-23 Published:2019-09-15 Online:2019-09-18
  • Contact: du xueke E-mail:46699644@qq.com

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摘要： 目的 探讨程序性坏死特异性抑制剂-1（Nec-1）对呼吸机相关性肺损伤（VILI）大鼠的保护作用。方 法 将40只SD大鼠按随机数字表法分为自主呼吸组（C组）、正常潮气量（VT）组（N组）、高VT组（H组）和Nec-1组 共4组，每组10只。C组保持自主呼吸，N组、H组和Nec-1组分别给予8、40、40 mL/kg的VT行机械通气，Nec-1组在 机械通气开始时静脉给予Nec-1（1 mg/kg）。通气4 h后收集支气管肺泡灌洗液（BALF）和肺组织标本，测定BALF中 总细胞数、总蛋白水平、白细胞介素（IL）-6、IL-1β及肿瘤坏死因子-α（TNF-α）水平。测定肺组织湿/干质量比值（W/ D），HE染色观察肺组织病理学改变并进行肺组织评分；采用实时荧光定量反转录-聚合酶链反应（RT-qPCR）和蛋白 质免疫印迹（Western blot）分别检测肺组织中受体相互作用蛋白 1（RIPK1）、RIPK3 和核转录因子-κB p65（NF-κB p65）的mRNA及蛋白表达。结果 C组和N组大鼠BALF中和肺组织各指标差异均无统计学意义。H组大鼠BALF 中总细胞数、总蛋白、炎性因子 IL-6、IL-1β、TNF-α 水平、肺组织 W/D 比值、肺组织病理评分及肺组织内 RIPK1、 RIPK3、NF-κB p65 的 mRNA 和蛋白相对表达水平较 C 组和 N 组明显升高（P＜0.01）。与 H 组相比，Nec-1 组大鼠 BALF中和肺组织各指标水平下降（P＜0.01）。结论 RIPK1/RIPK3/NF-κB信号传导通路参与大鼠VILI，Nec-1对大 鼠VILI的肺脏具有一定的保护作用。

关键词: 机械通气, 呼吸机相关性肺损伤, 动物实验, 程序性坏死特异性抑制剂-1

Abstract: Objective To investigate the protective effect of necrostatin-1 on ventilator - induced lung injury in rats. Methods Forty SD rats were randomly divided into four groups (n=10 for each group), spontaneous respiration group (group C), normal tidal volume (VT) group (group N), high VT group (group H) and necrostatin-1 group (group Nec-1). All rats were intubated after tracheotomy. Group C maintained spontaneous breathing, group N, group H and group Nec-1 were given mechanical ventilation with respective tidal volume (8 mL/kg, 40 mL/kg and 40 mL/kg). The group Nec-1 was injected Nec- 1 intravenously (1 mg/kg) before mechanical ventilation. After 4 hours of ventilation, samples of bronchoalveolar lavage fluid (BALF) and lung tissues were harvested. The infiltrated total cells, the total protein, the amount of interleukin (IL-6, IL-1β) and tumor necrosis factor - α (TNF-α) in alveolar lavage fluid were measured. The wet/dry weight ratio (W/D) of lung tissues was calculated. The histopathological changes of lung tissues were observed. The mRNA and protein expressions of receptor interacting protein 1 (RIPK1), RIPK3 and nuclear transcription factor -κB p65 (NF-κB p65) in lung tissues were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot assay respectively. Results There were no significant differences in related indexes in alveolar lavage fluid and lung tissue between group C and group N. Compared with those in group C and group N, values were increased in group H including the infiltrated total cells, the total protein and the amount of IL-6, IL-1β and TNF-α in alveolar lavage fluid, the wet/dry weight ratio and the histopathological score of lung tissues, the mRNA and protein expression levels of RIPK1, RIPK3 and NF-κB p65 in lung tissues (P＜0.01). In group Nec-1 the related indexes in alveolar lavage fluid and lung tissue were significantly lower than those of group H (P＜0.01). Conclusion RIPK1/RIPK3/NF-κB signaling pathway is involved in rat ventilatorinduced lung injury, and Nec-1 plays a protective role in rat lungs of ventilator-induced lung injury.

Key words: mechanical ventilation, ventilator-induced lung injury, animal experimentation, necrostatin-1