GABA信号通路对脓毒症大鼠急性肺损伤内质网应激和线粒体自噬的影响

doi:10.11958/20231352

天津医药 ›› 2024, Vol. 52 ›› Issue (7): 733-737.doi: 10.11958/20231352

GABA信号通路对脓毒症大鼠急性肺损伤内质网应激和线粒体自噬的影响

钟敏¹(), 施震²^,^∆(), 周劲松², 李晋杰²

1 湖北中医药大学第一临床医学院（邮编430000）
2 湖北省中医院（光谷院区）疼痛科

收稿日期:2023-09-05 修回日期:2023-11-22 出版日期:2024-07-15 发布日期:2024-07-11
通讯作者: ^∆E-mail：anbair3359@163.com
作者简介:钟敏（1994），女，硕士在读，主要从事急慢性疼痛的中西医临床方面研究。E-mail：isk1ud4x@163.com
基金资助:
湖北省卫生健康委2019-2020年度科研立项项目(WJ2019F162)

Effects of GABA signaling pathway on endoplasmic reticulum stress and mitochondrial autophagy in septic rats with acute lung injury

ZHONG Min¹(), SHI Zhen²^,^∆(), ZHOU Jinsong², LI Jinjie²

1 The First Clinical College of Hubei University of Traditional Chinese Medicine, Wuhan 430000, China
2 Department of Pain, Hubei Hospital of Traditional Chinese Medicine

Received:2023-09-05 Revised:2023-11-22 Published:2024-07-15 Online:2024-07-11
Contact: ^∆E-mail：anbair3359@163.com

钟敏, 施震, 周劲松, 李晋杰. GABA信号通路对脓毒症大鼠急性肺损伤内质网应激和线粒体自噬的影响[J]. 天津医药, 2024, 52(7): 733-737.

ZHONG Min, SHI Zhen, ZHOU Jinsong, LI Jinjie. Effects of GABA signaling pathway on endoplasmic reticulum stress and mitochondrial autophagy in septic rats with acute lung injury[J]. Tianjin Medical Journal, 2024, 52(7): 733-737.

摘要/Abstract

摘要：

目的探讨γ-氨基丁酸（GABA）信号通路对脓毒症大鼠急性肺损伤（ALI）内质网应激（ERS)和线粒体自噬的影响。方法 SD大鼠分为CON组、Model组、GABA信号通路激活剂巴氯芬组（巴氯芬组）、GABA信号通路抑制剂荷包牡丹碱（BIC）组（BIC组），每组6只。巴氯芬组腹腔注射5 mg/kg的巴氯芬，BIC组腹腔注射1 mg/kg BIC，每日1次，连续处理2周。CON组、Model组腹腔注射等量生理盐水。酶联免疫吸附试验检测血清中超氧化物歧化酶（SOD）、丙二醛（MDA）水平及支气管肺泡灌洗液（BALF）中细胞色素C（Cyt.C）、烟酰胺腺嘌呤二核苷酸磷酸（NADPH）水平；透射电子显微镜（TEM）观察肺组织细胞超微形态；HE染色观察肺组织病理形态；TUNEL染色观察肺组织细胞凋亡；Western blot检测肺组织GABAAR、GRP78、CHOP蛋白表达。结果与Model组相比，巴氯芬组肺肿胀、充血、炎性细胞浸润现象改善，肺损伤评分、MDA含量、凋亡指数、Cyt.C和NADPH水平、GRP78和CHOP蛋白水平降低（P<0.05），吞噬线粒体的自噬性空泡数量、SOD含量、GABAAR蛋白水平增加（P<0.05），而BIC组以上指标结果与巴氯芬组趋势相反。结论活化GABA信号通路可改善脓毒症大鼠ALI。

关键词: 脓毒症, 急性肺损伤, 内质网应激, 线粒体自噬, γ-氨基丁酸

Abstract:

Objective To investigate the effect of γ-aminobutyric acid (GABA) signaling pathway on endoplasmic reticulum (ER) stress and mitochondrial autophagy in septic rats with acute lung injury (ALI). Methods SD rats were randomly grouped into the control (CON) group, the model group, the GABA signaling pathway activator Baclofen group (the Baclofen group), the GABA signaling pathway inhibitor dicentrine group (the BIC group), with 6 rats in each group. The Baclofen group received intraperitoneal injection of 5 mg/kg Baclofen, and the BIC group received intraperitoneal injection of 1 mg/kg BIC, once a day, for two consecutive weeks. The CON group and the model group were injected with an equal amount of physiological saline via intraperitoneal injection. Enzyme linked immunosorbent assay was applied to detect serum levels of superoxide dismutase (SOD) and malondialdehyde (MDA) and levels of cytochrome C (Cyt.C) and Nicotinamide adenine dinucleotide phosphate (NADPH) in bronchoalveolar lavage fluid (BALF). Transmission electron microscopy (TEM) was applied to observe the ultrastructure of lung tissue cells. HE staining was applied to observe the pathological morphology of lung tissue. TUNEL staining was applied to observe the apoptosis of lung tissue. Western blot assay was applied to detect expression levels of GABAAR, GRP78 and CHOP proteins in lung tissue. Results Compared with the model group, the lung swelling, congestion and inflammatory cell infiltration were improved in the Baclofen group, and the lung injury score, MDA content, apoptosis index, Cyt.C and NADPH levels, GRP78, and CHOP protein levels were reduced (P<0.05). The number of autophagic vacuoles in phagocytic mitochondria, SOD content and GABAAR protein level were increased (P<0.05), however, the trend of above indicators in the BIC group was opposite to that in the Baclofen group. Conclusion Up-regulation of GABA signaling pathway may have an improvement effect on ALI in sepsis rats.

Key words: sepsis, acute lung injury, endoplasmic reticulum stress, mitophagy, gamma-aminobutyric acid

中图分类号:

R563

图/表 8

表1 各组大鼠SOD、MDA水平的比较（n=6，$\bar{x}±s$）

Tab.1 Comparison of SOD and MDA contents of rats between the four groups

组别	SOD/（U/mL）	MDA/（nmol/L）
CON组	1.96±0.26	9.83±0.83
Model组	0.77±0.08^a	41.67±4.53^a
巴氯芬组	1.45±0.22^b	17.63±1.83^b
BIC组	0.41±0.04^b	69.48±7.58^b
F	92.779^＊＊	211.485^＊＊

表2 各组大鼠Cyt.C和NADPH水平比较（n=6，$\bar{x}±s$）

Tab.2 Comparison of Cyt.C and NADPH levels between the four groups

组别	Cyt.C/（ng/L）	NADPH/（nmol/L）
CON组	10.56±1.04	102.09±11.13
Model组	25.98±3.12^a	151.76±16.23^a
巴氯芬组	13.97±1.45^b	111.25±11.21^b
BIC组	42.72±5.32^b	195.52±22.18^b
F	122.981^＊＊	43.661^＊＊

图1 TEM观察肺组织细胞线粒体自噬（×20 000）

Fig.1 Mitochondrial autophagy of lung tissue observed by TEM （×20 000）

图2 HE染色检测肺组织损伤(×200)

Fig.2 Lung tissue injury detected by HE staining （×200）

表3 各组大鼠肺损伤评分和细胞凋亡指数比较（n=6，$\bar{x}±s$）

Tab.3 Comparison of lung injury score and apoptosis index of rats between the four groups

组别	肺损伤评分/分	凋亡指数/%
CON组	0.02±0.01	5.15±0.62
Model组	2.57±0.32^a	31.28±4.13^a
巴氯芬组	0.44±0.04^b	12.19±1.63^b
BIC组	3.49±0.51^b	45.32±5.63^b
F	183.446^＊＊	154.652^＊＊

图3 TUNEL染色检测大鼠肺组织细胞凋亡（×400）

Fig.3 Apoptosis in rat lung tissue detected by TUNEL staining（×400）

图4 Western blot检测GABAAR、GRP78、CHOP蛋白水平

Fig.4 The protein levels of GABAAR, GRP78 and CHOP detected by Western blot assay

表4 各组大鼠肺组织GABAAR、GRP78、CHOP蛋白表达比较（n=6，$\bar{x}±s$）

Tab.4 Comparison of GABAAR, GRP78 and CHOP protein expression in lung tissue between four groups of rats

组别	GABAAR	GRP78	CHOP
CON组	0.96±0.09	0.48±0.03	0.79±0.10
Model组	0.38±0.02^a	1.44±0.15^a	1.56±0.20^a
巴氯芬组	0.92±0.08^b	0.53±0.05^b	0.85±0.09^b
BIC组	0.16±0.01^b	1.69±0.17^b	1.92±0.28^b
F	252.747^＊＊	168.774^＊＊	53.509^＊＊

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GABA信号通路对脓毒症大鼠急性肺损伤内质网应激和线粒体自噬的影响

Effects of GABA signaling pathway on endoplasmic reticulum stress and mitochondrial autophagy in septic rats with acute lung injury

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