隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制

doi:10.3969/j.issn.1006-9771.2022.05.009

《中国康复理论与实践》 ›› 2022, Vol. 28 ›› Issue (5): 544-551.doi: 10.3969/j.issn.1006-9771.2022.05.009

隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制

周小珏¹,冯婧¹,庞日朝²,刘捷³,张安仁⁴()

1.成都中医药大学养生康复学院,四川成都市 610075
2.西部战区总医院,四川成都市 610036
3.加拿大不列颠哥伦比亚大学,加拿大温哥华 V6T1Z4
4.同济大学附属上海市第四人民医院康复医学科,上海市 200434

收稿日期:2022-02-17 修回日期:2022-03-29 出版日期:2022-05-25 发布日期:2022-06-10
通讯作者: 张安仁 E-mail:anren0124@163.com
作者简介:周小珏(1996-),女,汉族,四川内江市人,硕士研究生,主要研究方向：脊髓损伤康复。
基金资助:
国家自然科学基金面上项目(8197151951)

Every-other-day fasting attenuated inflammation in rats after spinal cord injury via the aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB signaling pathway

ZHOU Xiaojue¹,FENG Jing¹,PANG Rizhao²,LIU Jie³,ZHANG Anren⁴()

1. School of Health Rehabilitation, Chengdu University of Tranditional Chinese Medicine, Chengdu, Sichuan 610075, China
2. Western Theater Command General Hospital, Chengdu, Sichuan 610036, China
3. University of British Columbia, Vancouver, V6T1Z4, Canada
4. Department of Rehabilitation Medicine, Shanghai Fourth People's Hospital Affiliated to Tongji University, Shanghai 200434, China

Received:2022-02-17 Revised:2022-03-29 Published:2022-05-25 Online:2022-06-10
Contact: ZHANG Anren E-mail:anren0124@163.com
Supported by:
National Natural Science Foundation of China (General)(8197151951)

摘要/Abstract

摘要：

目的探讨芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB (AHR/SOCS2/NF-κB)信号通路在隔日限食(EODF)疗法减轻脊髓损伤炎症反应中的作用。方法 36只雄性Sprague-Dawley大鼠随机分为假手术组、模型组和EODF组,每组12只。后两组复制C₅脊髓半侧钳夹模型。术后EODF组予24 h禁食和摄食交替,不限水,共2周。术前和术后1 d、7 d、14 d行梳理试验。术后14 d取各组脊髓组织,每组4只行HE染色观察病理损伤,4只采用Western blotting检测AHR、SOCS2和NF-κB水平,4只采用逆转录实时定量聚合酶链反应检测AHR、SOCS2、核因子κB抑制因子α (IκBα)、NF-κB、肿瘤坏死因子-α (TNF-α)、白细胞介素-6 (IL-6)的mRNA水平。结果与假手术组相比,另两组各时间点梳理试验评分降低(P < 0.05),出现空腔、水肿、结构紊乱等病理改变;与模型组相比,EODF组术后14 d梳理试验评分增加(P < 0.05),损伤减轻。与模型组相比,EODF组AHR、SOCS2 mRNA和蛋白表达增高(P < 0.05),NF-κB mRNA和蛋白表达降低(P < 0.05),IκBα mRNA表达增高(P < 0.05),TNF-α、IL-6 mRNA表达降低(P < 0.05)。结论 EODF可促进脊髓损伤大鼠患侧前肢运动功能恢复,减轻损伤和炎症反应,可能与激活AHR/SOCS2/NF-κB信号通路有关。

关键词: 脊髓损伤, 炎症, 隔日限食, 芳香烃受体, 细胞因子信号传导抑制因子2, 核转录因子-κB, 大鼠

Abstract:

Objective To explore the role of aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB (AHR/SOCS2/NF-κB) signaling in the reduction of spinal cord injury (SCI) inflammation with every-other-day fasting (EODF). Methods A total of 36 healthy male Sprague-Dawley rats were randomly divided into sham group (n = 12), model group (n = 12) and EODF group (n = 12). The latter two groups were established C₅ spinal cord hemi-clamp model. The EODF group received EODF, namely fasting and feeding per 24 hours (water free), for two weeks after operation. They were assessed with Grooming Test before operation, and one, seven and 14 days after operationt, respectively. Pathological injury of spinal cord was observed with HE staining 14 days after operation, while the expression of AHR, SOCS2 and NF-κB proteins were detected with Western blotting, and the mRNA of AHR, SOCS2, inhibitor α of NF-κB (IκBα), NF-κB and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) were measured with reverse transcription real-time quantitative polymerase chain reaction. Results Compared with the sham group, the Grooming Test score decreased in the other two groups (P < 0.05), and cavity, edema and structural disorder appeared in the spinal cord. Compared with the model group, the pathology of EODF group improved, and the Grooming Test score increased (P < 0.05). The mRNA and protein expression of AHR and SOCS2 increased (P < 0.05), the mRNA and protein expression of NF-κB decreased (P < 0.05), the mRNA expression of IκBα increased (P < 0.05), and the mRNA expression of TNF-α and IL-6 decreased (P < 0.05). Conclusion EODF may promote the recovery of motor function of forelimb in rats with SCI, and relieve damage and inflammation, which may associate with the activation of AHR/SOCS2/NF-κB signaling pathway.

Key words: spinal cord injury, inflammation, every-other-day fasting, aryl hydrocarbon receptor, suppressor of cytokine signaling 2, nuclear transcription factor-κB, rats

中图分类号:

R651.2

周小珏,冯婧,庞日朝,刘捷,张安仁. 隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制[J]. 《中国康复理论与实践》, 2022, 28(5): 544-551.

ZHOU Xiaojue,FENG Jing,PANG Rizhao,LIU Jie,ZHANG Anren. Every-other-day fasting attenuated inflammation in rats after spinal cord injury via the aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB signaling pathway[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(5): 544-551.

图/表 5

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表2

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表3

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组别	n	术前	术后1 d	术后7 d	术后14 d	F值	P值
假手术组	12	4.79±0.50	2.42±0.76	3.67±0.86	4.13±0.74	22.667	< 0.001
模型组	12	4.83±0.39	0.58±0.51^a	0.83±0.58^a	1.17±0.39^a	213.067	< 0.001
EODF组	12	4.58±0.51	0.50±0.52^a	0.92±0.67^a	1.67±0.49^a,b	132.724	< 0.001
F值		0.972	37.682	61.458	95.517
P值		0.389	< 0.001	< 0.001	< 0.001

组别	n	AHR	SOCS2	NF-κB	IκBα	TNF-α	IL-6
假手术组	4	1.231±0.237	1.265±0.463	1.000±0.302	1.951±0.488	0.865±0.103	0.475±0.031
模型组	4	0.576±0.207^a	0.502±0.015^a	1.887±0.464^a	1.054±0.424^a	1.575±0.120^a	0.853±0.136^a
EODF组	4	1.412±0.352^b	1.233±0.408^b	0.906±0.133^b	2.347±0.347^b	0.819±0.264^b	0.435±0.136^b
F值		10.380	5.857	10.840	9.771	22.760	16.770
P值		0.005	0.024	0.004	0.006	< 0.001	< 0.001

组别	n	AHR	SOCS2	NF-κB
假手术组	4	0.897±0.073	0.616±0.132	0.744±0.112
模型组	4	0.630±0.059^a	0.253±0.067^a	1.199±0.233^a
EODF组	4	0.961±0.189^b	0.723±0.282^b	0.857±0.144^b
F值		8.318	7.204	7.653
P值		0.009	0.014	0.011

隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制

Every-other-day fasting attenuated inflammation in rats after spinal cord injury via the aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB signaling pathway

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