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[低浓度氢气吸入对小鼠哮喘及睡眠功能的影响]

[Effects of Low Concentration Hydrogen Inhalation on Asthma and Sleep Function in Mice].

作者信息

Zhao Yue, Wei Lu, Zhang Xue-Wei, Zhu Li, Du Jun-Rong

机构信息

Department of Pharmacology, West China School of Pharmacy, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Mar;51(2):219-224. doi: 10.12182/20200360103.

Abstract

OBJECTIVE

This study was designed to investigate the effects of low concentration hydrogen inhalation on asthma and sleep function in mice and the potential mechanism.

METHODS

In the asthma experiment, BALB/c mice were randomly divided into normal control group, asthma model group and hydrogen treatment group. After establishing ovalbumin (OVA)-induced asthma model, the hydrogen treatment group mice were treated by inhalation of hydrogen (24-26 mL/L per day) for 7 consecutive days, and the normal control group and asthma model group mice received similar treatment by inhalation of air. The levels of interleukin (IL)-4, IL-13, and interferon-γ (IFN-γ) in bronchoalveolar lavage fluid (BALF) were measured by commercially available ELISA kits. The levels of malondialdehyde (MDA) and glutathione (GSH), as well as the activity of superoxide dismutase (SOD) in lung tissue were detected by colorimetric assays. The pathological changes in lung tissue were assessed by HE staining. In the sleep experiment, ICR mice were randomly divided into blank control group and 1 d, 3 d, 5 d hydrogen treatment groups and diazepam group. The effects of inhalation of 24-26 mL/L per day hydrogen on the sleep duration induced by intraperitoneal injection of upper-threshold dose of sodium pentobarbital and the sleep latency in response to subthreshold dose were evaluated.

RESULTS

In the asthma experiment, the asthma model group showed higher levels of IL-4 and IL-13 ( <0.05) and lower levels of IFN-γ ( <0.001) in BALF, as compared to the normal control group. The content of MDA in lung tissue was also significantly increased ( <0.01), companied by a decreased GSH concentration ( <0.05) and a mildly reduced SOD activity ( >0.05). Compared to the asthma model group, treatment with hydrogen significantly decreased the levels of IL-4 and IL-13 and increased the level of IFN-γ in BALF ( <0.05). Moreover, without alteration of the MDA production ( >0.05), hydrogen inhalation greatly increased GSH level and restored the SOD activity ( <0.05) in lung tissue. Additionally, the HE staining data showed that the hydrogen treatment attenuated the pulmonary histopathological changes. In the sleep experiment, compared with the blank control group, the sleep latency was significantly shorter ( <0.05) and the sleep duration was longer ( <0.001) in all the hydrogen treatment groups after receiving an upper-threshold dose of sodium pentobarbital. Meanwhile, in all the hydrogen treatment groups, the sleep latency was significantly longer ( <0.001) and the sleep duration was shorter ( <0.001) when compared to the diazepam group. Compared with the blank control group, after intraperitoneal injection of a subthreshold dose of sodium pentobarbital, the sleep latency was significantly increased in both 1 d and 5 d hydrogen treatment groups, and there was no significant difference as compared to the diazepam group. In the 3 d hydrogen treatment group, the sleep latency was only slightly increased ( >0.05), which was significantly lower than that of the diazepam group ( <0.05).

CONCLUSION

Low concentration hydrogen inhalation could alleviate OVA-induced asthma in mice, and the mechanism might be related to the anti-oxidative and anti-inflammatory effects of hydrogen. Also, low concentration hydrogen inhalation could improve sleep function in mice.

摘要

目的

本研究旨在探讨低浓度氢气吸入对小鼠哮喘及睡眠功能的影响及其潜在机制。

方法

在哮喘实验中,将BALB/c小鼠随机分为正常对照组、哮喘模型组和氢气治疗组。建立卵清蛋白(OVA)诱导的哮喘模型后,氢气治疗组小鼠连续7天每天吸入氢气(24 - 26 mL/L),正常对照组和哮喘模型组小鼠通过吸入空气进行类似处理。使用市售ELISA试剂盒检测支气管肺泡灌洗液(BALF)中白细胞介素(IL)-4、IL-13和干扰素-γ(IFN-γ)的水平。通过比色法检测肺组织中丙二醛(MDA)和谷胱甘肽(GSH)的含量以及超氧化物歧化酶(SOD)的活性。通过HE染色评估肺组织的病理变化。在睡眠实验中,将ICR小鼠随机分为空白对照组、1天、3天、5天氢气治疗组和地西泮组。评估每天吸入24 - 26 mL/L氢气对腹腔注射阈上剂量戊巴比妥钠诱导的睡眠时间和阈下剂量反应的睡眠潜伏期的影响。

结果

在哮喘实验中,与正常对照组相比,哮喘模型组BALF中IL-4和IL-13水平较高(<0.05),IFN-γ水平较低(<0.001)。肺组织中MDA含量也显著增加(<0.01),同时GSH浓度降低(<0.05),SOD活性轻度降低(>0.05)。与哮喘模型组相比,氢气治疗显著降低了BALF中IL-4和IL-13水平,并提高了IFN-γ水平(<0.05)。此外,吸入氢气在不改变MDA产生(>0.05)的情况下,显著提高了肺组织中GSH水平并恢复了SOD活性(<0.05)。另外,HE染色数据显示氢气治疗减轻了肺部组织病理学变化。在睡眠实验中,与空白对照组相比,所有氢气治疗组在接受阈上剂量戊巴比妥钠后睡眠潜伏期显著缩短(<0.05),睡眠时间延长(<0.001)。同时,与地西泮组相比,所有氢气治疗组睡眠潜伏期显著延长(<0.001),睡眠时间缩短(<0.001)。与空白对照组相比,腹腔注射阈下剂量戊巴比妥钠后,1天和5天氢气治疗组睡眠潜伏期显著延长,与地西泮组相比无显著差异。在3天氢气治疗组中,睡眠潜伏期仅略有增加(>0.05),显著低于地西泮组(<0.05)。

结论

低浓度氢气吸入可减轻OVA诱导的小鼠哮喘,其机制可能与氢气的抗氧化和抗炎作用有关。此外,低浓度氢气吸入可改善小鼠的睡眠功能。

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