中国全科医学 ›› 2024, Vol. 27 ›› Issue (06): 739-746.DOI: 10.12114/j.issn.1007-9572.2023.0270

• 论著·中医·中西医结合研究 • 上一篇    下一篇

甘草预防气管插管致呼吸道损伤的有效成分筛选

张杰1,2,3, 丁声双3, 郭敏4, 薛阳1,2, 徐紫清1,2, 侯怀晶1,2, 薛建军1,2,*()   

  1. 1730050 甘肃省兰州市,甘肃省中医院麻醉科
    2730050 甘肃省兰州市,甘肃省中西医结合麻醉临床研究中心
    3730000 甘肃省兰州市,甘肃中医药大学第一临床医学院
    4730050 甘肃省兰州市,甘肃省中医院
  • 收稿日期:2023-04-03 修回日期:2023-06-29 出版日期:2024-02-20 发布日期:2023-11-21
  • 通讯作者: 薛建军

  • 作者贡献:张杰和薛建军构思并设计了实验,指导研究实施与修改稿件;张杰、丁声双、郭敏、薛阳负责实验实施和血清学指标检测;张杰和郭敏撰写论文初稿。丁声双和徐紫清进行HE染色和免疫组化;丁声双和侯怀晶统计分析数据;所有作者审阅并批准了最终稿件。
  • 基金资助:
    甘肃省科技计划资助项目-临床医学研究中心建设(20JR10RA435); 兰州市人才创新创业计划(2018-RC-93)

Effective Fraction Screening of Glycyrrhiza Radix on Preventing Respiratory Tract Injury Induced by Endotracheal Intubation

ZHANG Jie1,2,3, DING Shengshuang3, GUO Min4, XUE Yang1,2, XU Ziqing1,2, HOU Huaijing1,2, XUE Jianjun1,2,*()   

  1. 1Department of Anesthesiology, Gansu Provincial Hospital of TCM, Lanzhou 730050, China
    2Gansu Province Clinical Research Center for Integrative Anaesthesiology, Lanzhou 730050, China
    3The First Clinical Medical College, Gansu University of Chinese Medicine, Lanzhou 730000, China
    4Gansu Provincial Hospital of TCM, Lanzhou 730050, China
  • Received:2023-04-03 Revised:2023-06-29 Published:2024-02-20 Online:2023-11-21
  • Contact: XUE Jianjun

摘要: 背景 术后咽喉痛(POST)是全身麻醉后常见的轻微但令人不悦的术后并发症之一。研究发现甘草能有效降低POST的发生率,但其有效成分尚不清楚。 目的 筛选甘草预防气管插管致呼吸道损伤的有效成分。 方法 2021年4月—2022年7月将60只SPF级Wistar大鼠依据随机数字表法分为对照组、气管插管组、利多卡因组、总多糖组、总皂苷组、总黄酮组,每组10只。除对照组外,其余组制备气管插管模型,插管前各治疗组按1 mL/100 g(按体质量计)分别经口喷入1%利多卡因、总多糖、总皂苷、总黄酮,浸润悬雍垂及软腭周围组织,对照组和气管插管组经口喷入等体积0.9%氯化钠溶液。机械通气2 h后拔除气管导管,深麻醉下收集大鼠咽部黏膜组织和血液标本,通过苏木精-伊红染色法(HE染色)和免疫组化(IHC)观察大鼠咽部黏膜病理变化和Toll样受体2(TLR2)、Toll样受体4(TLR4)表达水平;酶联免疫吸附测定法(ELISA)检测大鼠血清炎性因子[肿瘤坏死因子α(TNF-α)、白介素(IL)-2、IL-4、IL-10]、氧化应激[超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC)]及应激激素[皮质醇(Cor)、肾上腺素(E)和去甲肾上腺素(NE)]表达水平。 结果 HE染色显示气管插管组可见黏膜脱落,黏膜下结构严重破坏并伴有大量炎性细胞浸润,总皂苷组大鼠咽部黏膜组织破坏较轻,炎性细胞明显减少。ELISA结果显示:与对照组相比,气管插管组大鼠血清中的TNF-α、IL-2水平升高,IL-4、IL-10水平降低(P<0.05);与气管插管组相比,总皂苷组大鼠血清中TNF-α和IL-2水平降低,IL-4和IL-10水平升高(P<0.05)。与对照组相比,气管插管组大鼠血清中MDA水平升高,SOD和T-AOC水平下降(P<0.05);与气管插管组相比,总皂苷组和总黄酮组大鼠血清中MDA水平降低,SOD和T-AOC水平升高(P<0.05)。与对照组相比,气管插管组大鼠血清Cor、E和NE水平升高(P<0.05);与气管插管组相比,总皂苷组大鼠血清Cor、E和NE水平降低(P<0.05)。IHC结果显示:气管插管组大鼠咽部黏膜组织TLR2和TLR4表达的平均光密度(AOD)高于对照组(P<0.05)。与气管插管组相比,总皂苷组大鼠咽部黏膜组织TLR4表达的AOD降低(P<0.05)。 结论 甘草预防全身麻醉气管插管致呼吸道损伤的主要有效成分为总皂苷。

关键词: 甘草, 气管插管, 麻醉, 通气,机械, 呼吸道损伤, 有效成分, 总皂苷

Abstract:

Background

Postoperative sore throat (POST) is one of the common mild but unpleasant postoperative complications after general anaesthesia. There is evidence that Glycyrrhiza Radix can effectively reduce the incidence of POST, but its effective fraction remains unclear.

Objective

To screen the effective fraction of Glycyrrhiza Radix on preventing respiratory tract injury induced by endotracheal intubation.

Methods

From April 2021 to July 2022, 60 SPF-grade Wistar rats were randomized into the control group, endotracheal intubation group, lidocaine group, total polysaccharides group, total saponins group, total flavonoids group based on random number table method, with 10 rats in each group. The endotracheal intubation model was prepared for all groups except the control group. Before intubation, each treatment group was sprayed with a solution consisting of 1% lidocaine, total polysaccharides, total saponins, and total flavonoids at 1 mL/100 g (by body weight) to permeate the uvula and peri-soft palate tissue of rats, respectively. The control and endotracheal intubation groups were sprayed with an equal volume of 0.9% saline. After 2 hours of mechanical ventilation, the endotracheal tube was removed, and the pharyngeal mucosa tissues and blood samples of rats were collected under deep anesthesia. Pathological changes of rat pharyngeal mucosa and the expression levels of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) were observed by hematoxylin-eosin staining (HE staining) and immunohistochemistry (IHC) ; the expression levels of inflammatory cytokines including serum tumor necrosis factor-α (TNF-α) , interleukin-2 (IL-2) , interleukin-4 (IL-4) , interleukin-10 (IL-10) , oxidative stress including superoxide dismutase (SOD) , malondialdehyde (MDA) , total antioxidant capacity (T-AOC) , and stress hormones including cortisol (Cor) , epinephrine (E) , and norepinephrine (NE) were measured by enzyme-linked immunosorbent assay (ELISA) .

Results

HE staining showed mucosal shedding and severe damage of submucosal structure accompanied by a large number of inflammatory cells infiltrated in the endotracheal intubation group; while milder damage of mucosal structure and less inflammatory cells were significantly reduced in the total saponins group. ELISA results showed that compared with the control group, the levels of TNF-α and IL-2 in the serum of rats increased significantly, and the levels of IL-4 and IL-10 decreased significantly in the endotracheal intubation group (P<0.05) ; compared with the endotracheal intubation group, the serum levels of TNF-α and IL-2 decreased, and the levels of IL-4 and IL-10 increased in the total saponins group (P<0.05) . Compared with the control group, the MDA level in the serum of rats was significantly increased, and the SOD and T-AOC levels were significantly decreased in the endotracheal intubation group (P<0.05) ; compared with the endotracheal intubation group, the MDA level decreased and the levels of SOD and T-AOC increased in the total saponins and flavonoids groups (P<0.05) . Compared with the control group, serum Cor, E, and NE levels significantly increased in the endotracheal intubation group (P<0.05) ; compared with the tracheal intubation group, serum Cor, E and NE levels were decreased in the total saponins group rats (P<0.05) . The IHC results showed that the average optical density (AOD) expressed by TLR2 and TLR4 in the endotracheal intubation group was higher than the control group (P<0.05) . The AOD expressed by TLR4 in the pharyngeal mucosal tissue of rats in the total saponins group was significantly lower than that of the endotracheal intubation group (P<0.05) .

Conclusion

Total saponin is the main effective fraction of Glycyrrhiza Radix for the prevention of respiratory tract injury caused by endotracheal intubation under general anesthesia.

Key words: Glycyrrhiza radix, Endotracheal intubation, Anesthesia, Ventilation, mechanical, Respiratory tract injury, Effective fraction, Total saponins