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

doi:10.11958/20231352

Abstract

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

CLC Number:

R563

Figures/Tables 8

References 23

[1]	CINAR I, SIRIN B, AYDIN P, et al. Ameliorative effect of gossypin against acute lung injury in experimental sepsis model of rats[J]. Life Sci, 2019, 221:327-334. doi:10.1016/j.lfs.2019.02.039.
[2]	LIU F, PENG W, CHEN J, et al. Exosomes derived from alveolar epithelial cells promote alveolar macrophage activation mediated by mirR-92a-3p in sepsis-induced acute lung injury[J]. Front Cell Infect Microbiol, 2021,11:646546. doi:10.3389/fcimb.2021.646546.
[3]	CHEN X, WANG Y, XIE X, et al. Heme oxygenase-1 reduces sepsis-induced endoplasmic reticulum stress and acute lung injury[J]. Mediators Inflamm, 2018,2018:9413876. doi:10.1155/2018/9413876.
[4]	JIANG L, XU L, ZHENG L, et al. Salidroside attenuates sepsis-associated acute lung injury through PPP1R15A mediated endoplasmic reticulum stress inhibition[J]. Bioorg Med Chem, 2022,71:116865. doi:10.1016/j.bmc.2022.116865.
[5]	CHEN H, LIN H, DONG B, et al. Hydrogen alleviates cell damage and acute lung injury in sepsis via PINK1/Parkin-mediated mitophagy[J]. Inflamm Res, 2021, 70(8):915-930. doi:10.1007/s00011-021-01481-y.
[6]	HUANG T, ZHANG Y, WANG C, et al. Propofol reduces acute lung injury by up-regulating gamma-aminobutyric acid type a receptors[J]. Exp Mol Pathol, 2019,110:104295. doi:10.1016/j.yexmp.2019.104295.
[7]	LI R, REN T, ZENG J. Mitochondrial coenzyme q protects sepsis-induced acute lung injury by activating PI3K/Akt/GSK-3β/mTOR pathway in rats[J]. Biomed Res Int, 2019,2019:5240898. doi:10.1155/2019/5240898.
[8]	SHEN K, WANG X, WANG Y, et al. miR-125b-5p in adipose derived stem cells exosome alleviates pulmonary microvascular endothelial cells ferroptosis via Keap1/Nrf2/GPX4 in sepsis lung injury[J]. Redox Biol, 2023,62:102655. doi:10.1016/j.redox.2023.102655.
[9]	周正, 宋美慧, 褚光辉, 等. GABA_(B)受体激动剂巴氯芬对小鼠空间识别记忆能力的影响[J]. 生命科学研究, 2022, 26(3):213-218.
	ZHOU Z, SONG M H, CHU G H, et al. Effect of GABAB receptor agonist baclofen on spatial cognition in mice[J]. Life Science Research, 2022, 26(3):213-218. doi:10.16605/j.cnki.1007-7847.2021.02.0123.
[10]	刘乃红, 王佃卿, 彭志锋. 荷包牡丹碱联合早期跑步运动对大鼠脑损伤恢复的协同效应[J]. 中国应用生理学杂志, 2022, 38(6):700-704.
	LIU N H, WANG D Q, PENG Z F. Synergistic effect of dicentime combined with early running on brain injury recovery in rats[J]. Chinese Journal of Applied Physiology, 2022, 38(6):700-704. doi:10.12047/j.cjap.6379.2022.127.
[11]	WANG Y, YANG Y, CHEN L, et al. Death-associated protein kinase 1 mediates ventilator-induced lung injury in mice by promoting alveolar epithelial cell apoptosis[J]. Anesthesiology, 2020, 133(4):905-918. doi:10.1097/ALN.0000000000003464.
[12]	虢强, 朱美意, 张杰, 等. 红景天苷调节CLP诱导的脓毒症小鼠肺纤维化及JAK2/STAT3的活化[J]. 中国免疫学杂志, 2022, 38(10):1196-1200.
	GUO Q, ZHU M Y, ZHANG J, et al. Salidroside regulates pulmonary fibrosis and JAK2/STAT3 activation in CLP-induced sepsis mice[J]. Chinese Journal of Immunology, 2022, 38(10):1196-1200. doi:10.3969/j.issn.1000-484X.2022.10.008.
[13]	张昊, 苏乾, 史佳, 等. 电针通过NLRP3/caspase-1通路介导的细胞焦亡途径减轻脓毒症大鼠急性肺损伤[J]. 中国中西医结合外科杂志, 2023, 29(6):814-818.
	ZHANG H, SU Q, SHI J, et al. Electroacupuncture attenuates acute lung injury induced by sepsis via NLRP3/caspase-1 pathway mediated pyrocytosis pathway[J]. Chinese Journal of Surgery of Integrated Traditional and Western Medicine, 2023, 29(6):814-818. doi:10.3969/j.issn.1007-6948.2023.06.017.
[14]	吴海涛, 周洪礼, 宋玉, 等. 穿心莲内酯对盲肠结扎穿孔诱导的脓毒症大鼠心肺功能和炎症损伤的保护作用[J]. 天津医药, 2022, 50(1):67-73.
	WU H T, ZHOU H L, SONG Y, et al. Protective effects of andrographolide on cardiopulmonary function and inflammatory damage in sepsis rats induced by cecal ligation and perforation[J]. Tianjin Med J, 2022, 50(1):67-73.doi:10.11958/20210462.
[15]	YOUSSEF N S, ELZAITONY A S, ABDEL BAKY N A. Diacerein attenuate LPS-induced acute lung injury via inhibiting ER stress and apoptosis: impact on the crosstalk between SphK1/S1P, TLR4/NFκB/STAT3,and NLRP3/IL-1β signaling pathways[J]. Life Sci, 2022,308:120915. doi:10.1016/j.lfs.2022.120915.
[16]	PAO H P, LIAO W I, TANG S E, et al. Suppression of endoplasmic reticulum stress by 4-PBA protects against hyperoxia-induced acute lung injury via up-regulating claudin-4 expression[J]. Front Immunol, 2021,12:674316. doi:10.3389/fimmu.2021.674316.
[17]	ZHANG Z K, ZHOU Y, CAO J, et al. Rosmarinic acid ameliorates septic-associated mortality and lung injury in mice via GRP78/IRE1α/JNK pathway[J]. J Pharm Pharmacol, 2021, 73(7):916-921. doi:10.1093/jpp/rgaa033.
[18]	AHMAD S, ZAKI A, MANDA K, et al. Vitamin-D ameliorates sepsis-induced acute lung injury via augmenting miR-149-5p and downregulating ER stress[J]. J Nutr Biochem, 2022,110:109130. doi:10.1016/j.jnutbio.2022.109130.
[19]	SANG A, WANG Y, WANG S, et al. Quercetin attenuates sepsis-induced acute lung injury via suppressing oxidative stress-mediated ER stress through activation of SIRT1/AMPK pathways[J]. Cell Signal, 2022,96:110363. doi:10.1016/j.cellsig.2022.110363.
[20]	WANG T, ZHANG J, WEI H, et al. Matrine-induced nephrotoxicity via GSK-3β/nrf2-mediated mitochondria-dependent apoptosis[J]. Chem Biol Interact, 2023,378:110492. doi:10.1016/j.cbi.2023.110492.
[21]	LIU W, LI C C, LU X, et al. Overexpression of transcription factor EB regulates mitochondrial autophagy to protect lipopolysaccharide-induced acute lung injury[J]. Chin Med J(Engl), 2019, 132(11):1298-1304. doi:10.1097/CM9.0000000000000243.
[22]	ZHAO R, WANG B, WANG D, et al. Oxyberberine prevented lipopolysaccharide-induced acute lung injury through inhibition of mitophagy[J]. Oxid Med Cell Longev, 2021,2021:6675264. doi:10.1155/2021/6675264.
[23]	JULIANA B, ALVIN T K, LINH A, et al. Pulmonary neuroendocrine cells secrete γ-Aminobutyric Acid to Induce Goblet Cell Hyperplasia in Primate Models[J]. Am J Respir Cell Mol Biol, 2019, 60(6):687-694. doi:10.1165/rcmb.2018-0179OC.

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^＊＊

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^＊＊

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^＊＊

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^＊＊

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^＊＊