The effect of sevoflurane on neuronal inflammation in rats with spinal cord injury by regulating the ROS/TXNIP/NLRP3 pathway

doi:10.11958/20251781

Abstract

Abstract:

Objective To investigate the effect of sevoflurane on neuroinflammation in rats with spinal cord injury by regulating the reactive oxygen species (ROS)/thioredoxin-interacting protein (TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) pathway. Methods Seventy-two rats were randomly divided into six groups using a random number table method, with 12 rats in each group: the model group (establishing the rat model of spinal cord injury), the sham operation group, the low-, medium- and high-concentration sevoflurane groups (1%, 2% and 4% sevoflurane, respectively) and the combination group (4% sevoflurane + 3.7 mg/kg reactive oxygen species (ROS) activator trimethylamine N-oxide). Spinal cord injury behavior (BBB) score and footprint imprint test were used to evaluate motor function. ELISA method was used to detect the levels of IL-18, TNF-α and IL-1β in spinal cord tissue. HE staining was used to detect pathological changes in spinal cord tissue. TUNEL method was used to detect cell apoptosis. In addition, the expression of TXNIP and NLRP3 mRNA were detected by qRT-PCR. ROS activity was detected by ROS kit. Western blot assay was used to detect the expression of ROS/TXNIP/NLRP3 pathway proteins. Results The sham operation group showed normal gait, normal cell morphology with orderly arrangement and intact spinal cord structure. The model group presented with toe dragging, severe spinal cord tissue injury and inflammatory cell infiltration. The BBB score was lower in the model group compared to that of the sham surgery group, while IL-18, TNF-α, IL-1β levels, apoptosis rate, ROS activity, TXNIP, NLRP3 mRNA and protein expression were increased (P<0.05). The above indexes were improved in the low-concentration, medium-concentration and high-concentration sevoflurane groups compared with those in the model group. However, the combined group reversed the improvement results of the high-concentration sevoflurane group. Conclusion Sevoflurane can alleviate neuroinflammation in rats with spinal cord injury by inhibiting the ROS/TXNIP/NLRP3 pathway.

Key words: sevoflurane, spinal cord injuries, reactive oxygen species, thioredoxins, NLR family, pyrin domain-containing 3 protein, neurons, inflammation

CLC Number:

R651.2

Figures/Tables 9

References 25

[1]	秦晓健, 李子阳, 周鑫, 等. 橄榄苦苷对脊髓损伤大鼠神经细胞凋亡的影响及可能机制[J]. 解剖科学进展, 2024, 30(2):190-192.
	QIN X J, LI Z Y, ZHOU X, et al. Effect of oleuropein on nerve cell apoptosis in rats with spinal cord injury and its possible mechanism[J]. Progress of Anatomical Sciences, 2024, 30(2):190-192. doi:10.16695/j.cnki.1006-2947.2024.02.021.
[2]	ZHANG H, PIAO M, GUO M, et al. MicroRNA-211-5p attenuates spinal cord injury via targeting of activating transcription factor 6[J]. Tissue Cell, 2021, 68:101459. doi:10.1016/j.tice.2020.101459.
[3]	周孝敏, 鱼丽萍, 李卓伦, 等. 夹脊电针刺激通过PKA/RhoA通路治疗大鼠急性脊髓损伤[J]. 神经解剖学杂志, 2023, 39(3):295-302.
	ZHOU X M, YU L P, LI Z L, et al. Treatment of acute spinal cord injury in rats with Jiaji electro-acupuncture stimulation via PKA/RhoA pathway[J]. Chin J Neuroanat, 2023, 39(3):295-302. doi:10.16557/j.cnki.1000-7547.2023.03.007.
[4]	LIU X M, XU M, ZHANG H, et al. Effects of electroacpuncture on urodynamics,intramedullary apoptosis and bidirectional regulation of neurotrophic factors in neurogenic bladder rats after supersacral spinal cord injury[J]. World J Acupunct Moxibustion, 2021, 31(2):121-128.
[5]	姜山, 吴超, 马玲. 夹脊穴电针调控TREM2介导的小胶质细胞活化促进脊髓损伤修复的机制研究[J]. 免疫学杂志, 2024, 40(10):761-766.
	JIANG S, WU C, MA L. Mechanism of electro-acupuncture at jiaji acupoint in regulating TREM2-mediated microglia activation to promote spinal cord injury repair[J]. J Immunol, 2024, 40(10):761-766. doi:10.13431/j.cnki.immunol.j.20240106.
[6]	LI R, HAN J, CHEN B, et al. Homeodomain interacting protein kinase 2-modified rat spinal astrocytes affect neurofunctional recovery after spinal cord injury[J]. Curr Neurovasc Res, 2022, 19(2):171-180. doi:10.2174/1567202619666220601111715.
[7]	LIANG T Y, PENG S Y, MA M, et al. Protective effects of sevoflurane in cerebral ischemia reperfusion injury:a narrative review[J]. Med Gas Res, 2021, 11(4):152-154. doi:10.4103/2045-9912.318860.
[8]	XU Q, XING H, WU J, et al. miRNA-141 induced pyroptosis in intervertebral disk degeneration by targeting ROS generation and activating TXNIP/NLRP3 signaling in nucleus pulpous cells[J]. Front Cell Dev Biol, 2020, 8:871. doi:10.3389/fcell.2020.00871.
[9]	陈勇, 范军朝, 庞红利, 等. 七氟醚后处理对脑缺血/再灌注损伤大鼠的脑保护作用及对ROS/TXNIP/NLRP3通路的影响[J]. 国际麻醉学与复苏杂志, 2021, 42(1):8-14.
	CHEN Y, FAN J Z, PANG H L, et al. Protective effect of sevoflurane postconditioning on cerebral ischemia/reperfusion injury in rats and its influence on the ROS/TXNIP/NLRP3 pathway[J]. Int J Anesth Resus, 2021, 42(1):8-14. doi:10.3760/cma.j.cn321761-20200828-00184.
[10]	杨光, 姚富, 陈永旺, 等. 依托咪酯对脊髓损伤大鼠的脑保护作用及其机制研究[J]. 中国现代医学杂志, 2022, 32(6):32-37.
	YANG G, YAO F, CHEN Y W, et al. Cerebral protective effect of etomidate in rats with spinal cord injury and its mechanism[J]. Chin J Modern Med, 2022, 32(6):32-37. doi:10.3969/j.issn.1005-8982.2022.06.006.
[11]	程亮亮, 田毅, 谭义文, 等. 基于PI3K/Akt/mTOR通路探究七氟醚麻醉对大鼠神经细胞凋亡的影响[J]. 西北药学杂志, 2024, 39(2):47-52.
	CHENG L L, TIAN Y, TAN Y W, et al. Effect of sevoflurane anesthesia on neuronal apoptosis in rats based on PI3K/Akt/mTOR pathway[J]. Northwest Pharm J, 2024, 39(2):47-52. doi:10.3969/j.issn.1004-2407.2024.02.008.
[12]	田新磊, 朱珊, 赵文锦, 等. 橙皮苷抑制ROS/NLRP3通路改善小鼠急性支气管炎的作用机制研究[J]. 中药新药与临床药理, 2023, 34(4):501-508.
	TIAN X L, ZHU S, ZHAO W J, et al. Research on the mechanism of hesperidin inhibiting ROS/NLRP3 pathway to improve acute bronchitis in mice[J]. Traditional Chinese Drug Research and Clinical Pharmacology, 2023, 34(4):501-508. doi:10.19378/j.issn.1003-9783.2023.04.010.
[13]	DUAN H Q, WU Q L, YAO X, et al. Nafamostat mesilate attenuates inflammation and apoptosis and promotes locomotor recovery after spinal cord injury[J]. CNS Neurosci Ther, 2018, 24(5):429-438. doi:10.1111/cns.12801.
[14]	黄乔, 李涛, 赵莉莉, 等. 脊髓损伤小鼠通过激活小胶质细胞MyD88通路促进神经元铁死亡[J]. 神经解剖学杂志, 2023, 39(1):71-78.
	HUANG Q, LI T, ZHAO L L, et al. Activating MyD88 signal pathway in the microglia promotes neuronal ferroptosis after spinal cord injury[J]. Chin J Neur, 2023, 39(1):71-78. doi:10.16557/j.cnki.1000-7547.2023.01.010.
[15]	史旭, 李瑞语, 张兵, 等. 小胶质细胞极化介导炎症反应在脊髓损伤中的作用[J]. 中国组织工程研究, 2023, 27(1):121-129.
	SHI X, LI R Y, ZHANG B, et al. Effect of inflammatory reaction mediated by microglia polarization in spinal cord injury[J]. Chin J Tissue Eng Res, 2023, 27(1):121-129. doi:10.12307/2022.731.
[16]	王领, 赵雪, 王金林. 七氟醚通过p38/MAPK信号通路抑制脂多糖诱导的肺泡巨噬细胞M1型极化[J]. 中国免疫学杂志, 2024, 40(9):1850-1855.
	WANG L, ZHAO X, WANG J L. Sevoflurane inhibits M1-type polarization of alveolar macrophages induced by lipopolysaccharide through p38/MAPK signaling pathway[J]. Chinese Journal of Immunology, 2024, 40(9):1850-1855. doi:10.3969/j.issn.1000-484X.2024.09.010.
[17]	WANG F, LI C, SHAO J, et al. Sevoflurane induces inflammation of microglia in hippocampus of neonatal rats by inhibiting Wnt/ β-Catenin/CaMKIV pathway[J]. J Pharmacol Sci, 2021, 146(2):105-115. doi:10.1016/j.jphs.2021.02.004.
[18]	ZHU Z, MA L. Sevoflurane induces inflammation in primary hippocampal neurons by regulating Hoxa5/Gm5106/miR-27b-3p positive feedback loop[J]. Bioengineered, 2021, 12(2):12215-12226. doi:10.1080/21655979.2021.2005927.
[19]	FAN H, TANG H B, CHEN Z, et al. Inhibiting HMGB1-RAGE axis prevents pro-inflammatory macrophages/microglia polarization and affords neuroprotection after spinal cord injury[J]. J Neuroinflammation, 2020, 17(1):295. doi:10.1186/s12974-020-01973-4.
[20]	周雨昕, 马勇, 吴承杰, 等. 基于NLRP3炎症小体途径探究脊髓康促进脊髓损伤大鼠修复的机制[J]. 中华中医药杂志, 2022, 37(9):5385-5389.
	ZHOU Y X, MA Y, WU C J, et al. Mechanism of Jisui Kang in promoting the repair of spinal cord injury rats based on the NLRP3 inflammasome pathway[J]. CJTCMP, 2022, 37(9):5385-5389.
[21]	王欣, 束传亮. miR-17-5p调控TXNIP/NLRP3信号通路对牙龈卟啉单胞菌LPS诱导的人牙龈成纤维细胞炎症反应的影响[J]. 中国病原生物学杂志, 2025, 20(4):460-465.
	WANG X, SHU C L. The effect of miR-17-5p on Porphyromonas gingivalis LPS-induced inflammatory response of human gingival fibroblasts by regulating the TXNIP/NLRP3 signaling pathway[J]. J Path Bio, 2025, 20(4):460-465. doi:10.13350/j.cjpb.250410.
[22]	朱晶惠, 魏栋敏, 任淑婷, 等. 靶向抑制NLRP3炎性小体在脊髓损伤中的研究进展[J]. 天津医药, 2023, 51(7):781-784.
	ZHU J H, WEI D M, REN S T, et al. Research progress of targeted inhibition of NLRP3 inflammasome in spinal cord injury[J]. Tianjin Med J, 2023, 51(7):781-784. doi:10.11958/20221718.
[23]	LUO T, ZHOU X, QIN M, et al. Corilagin restrains NLRP3 inflammasome activation and pyroptosis through the ROS/TXNIP/NLRP3 pathway to prevent inflammation[J]. Oxid Med Cell Longev, 2022, 2022:1652244. doi:10.1155/2022/1652244.
[24]	LIU Y, WU H, WANG T, et al. Paeonol reduces microbial metabolite α-hydroxyisobutyric acid to alleviate the ROS/TXNIP/NLRP3 pathway-mediated endothelial inflammation in atherosclerosis mice[J]. Chin J Nat Med, 2023, 21(10):759-774. doi:10.1016/S1875-5364(23)60506-0.
[25]	ZHAO Q, LIU G, DING Q, et al. The ROS/TXNIP/NLRP3 pathway mediates LPS-induced microglial inflammatory response[J]. Cytokine, 2024, 181(1):156677. doi:10.1016/j.cyto.2024.156677.

基因名称	引物序列（5′→3′）	产物大小/bp
TXNIP	上游：CCGAGCCAGCCAACTCAAGA	189
TXNIP	下游：AGACAGACACCCGCCCATCA	189
NLRP3	上游：GAATTAGACAACTGCAACCTCACG	195
NLRP3	下游：GTTTCAGCACTTCACAGAACATCAT	195
GAPDH	上游：GGTGGTCTCCTCTGACTTCAA	178
GAPDH	下游：GTTGCTGTAGCCAAATTCGTTGT	178

基因名称	引物序列（5′→3′）	产物大小/bp
TXNIP	上游：CCGAGCCAGCCAACTCAAGA	189
TXNIP	下游：AGACAGACACCCGCCCATCA	189
NLRP3	上游：GAATTAGACAACTGCAACCTCACG	195
NLRP3	下游：GTTTCAGCACTTCACAGAACATCAT	195
GAPDH	上游：GGTGGTCTCCTCTGACTTCAA	178
GAPDH	下游：GTTGCTGTAGCCAAATTCGTTGT	178

组别	BBB评分
组别	第7天	第14天
假手术组	21.00±0.00	21.00±0.00
模型组	5.23±0.57^a	6.89±0.69^a
低浓度七氟醚组	6.59±0.66^b	8.53±0.87^b
中浓度七氟醚组	7.92±0.81^bc	10.05±1.12^bc
高浓度七氟醚组	9.25±0.93^bcd	11.86±1.26^bcd
联合组	6.15±0.62^e	7.62±0.79^e
F	932.357^＊＊	416.942^＊＊

组别	BBB评分
组别	第7天	第14天
假手术组	21.00±0.00	21.00±0.00
模型组	5.23±0.57^a	6.89±0.69^a
低浓度七氟醚组	6.59±0.66^b	8.53±0.87^b
中浓度七氟醚组	7.92±0.81^bc	10.05±1.12^bc
高浓度七氟醚组	9.25±0.93^bcd	11.86±1.26^bcd
联合组	6.15±0.62^e	7.62±0.79^e
F	932.357^＊＊	416.942^＊＊

组别	IL-18/（ng/L）	TNF-α/（μg/L）	IL-1β/（ng/L）
假手术组	61.28±6.25	6.58±0.67	101.27±10.35
模型组	151.32±15.27^a	18.32±1.91^a	195.37±20.17^a
低浓度七氟醚组	121.65±12.33^b	15.47±1.63^b	167.83±17.14^b
中浓度七氟醚组	104.57±11.57^bc	12.56±1.27^bc	136.42±13.82^bc
高浓度七氟醚组	85.66±8.72^bcd	9.36±0.95^bcd	119.67±12.17^bcd
联合组	139.58±14.05^e	16.89±1.72^e	178.69±17.92^e
F	49.446^＊＊	61.216^＊＊	32.756^＊＊