富氢液通过增加自噬治疗大鼠神经病理性疼痛

doi:10.11958/20230698

天津医药 ›› 2024, Vol. 52 ›› Issue (3): 261-265.doi: 10.11958/20230698

富氢液通过增加自噬治疗大鼠神经病理性疼痛

何颖¹(), 张广华¹, 田立东¹, 于泳浩²^,^△()

1.天津医科大学朱宪彝纪念医院麻醉科、天津市内分泌研究所、国家卫生健康委员会激素与发育重点实验室、天津市代谢性疾病重点实验室（邮编300134）
2.天津医科大学总医院麻醉科

收稿日期:2023-05-16 修回日期:2023-08-18 出版日期:2024-03-15 发布日期:2024-03-13
通讯作者: ^△E-mail：yyu@tmu.edu.cn
作者简介:何颖（1990），女，医师，主要从事神经病理性疼痛方面研究。E-mail：dxbhy9004@126.com
基金资助:
国家自然科学基金资助项目(81372033);天津市卫生健康科技项目(TJWJ2021MS021);天津市医学重点学科（专科）建设项目(TJYXZDXK-032A)

Hydrogen-rich saline treated neuropathic pain in rats by increasing autophagy

HE Ying¹(), ZHANG Guanghua¹, TIAN Lidong¹, YU Yonghao²^,^△()

1. National Health Commission Key Laboratory of Hormones and Development, Tianjin Key Laboratory of Metabolic Diseases, Department of Anesthesiology, Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China
2. Department of Anesthesiology, Tianjin Medical University General Hospital

Received:2023-05-16 Revised:2023-08-18 Published:2024-03-15 Online:2024-03-13
Contact: >^△E-mail: yyu@tmu.edu.cn

何颖, 张广华, 田立东, 于泳浩. 富氢液通过增加自噬治疗大鼠神经病理性疼痛[J]. 天津医药, 2024, 52(3): 261-265.

HE Ying, ZHANG Guanghua, TIAN Lidong, YU Yonghao. Hydrogen-rich saline treated neuropathic pain in rats by increasing autophagy[J]. Tianjin Medical Journal, 2024, 52(3): 261-265.

摘要/Abstract

摘要：

目的评价自噬在富氢液治疗神经病理性疼痛中的作用。方法将鞘内置管成功的40只成年雄性SD大鼠随机分为5组：假手术组（S组）、神经病理性疼痛组（C组）、富氢液组（H组）、自噬抑制剂组（M组）和富氢液+自噬抑制剂组（HM组），各8只。采用坐骨神经慢性压迫法（CCI）制备大鼠神经病理性疼痛模型。M组和HM组于术后鞘内注射自噬抑制剂3-甲基腺嘌呤（3-MA）30 μg/kg，H组和HM组于术后腹腔注射富氢液（0.6 mmol/L）10 mL/kg，其他组鞘内/腹腔给予等量生理盐水，2次/d，连续7 d。于造模前1 d和造模后1、3、5、7和14 d（T₀—T₅）测定大鼠机械刺激缩足阈值（MWT）和热刺激缩足潜伏期（TWL）。取脊髓L4—L6节段，采用Western blot法检测自噬相关蛋白微管相关蛋白轻链3（LC3）Ⅱ、Beclin-1和p62蛋白的表达；并测定脊髓组织中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。结果与S组比较，C组T₂—T₅时MWT和TWL均降低，T₅时脊髓LC3Ⅱ、Beclin-1和p62蛋白表达水平升高，SOD活性降低，MDA含量增加（P<0.05）。与C组比较，H组T₂—T₅时MWT和TWL均升高，T₅时脊髓LC3Ⅱ和Beclin-1蛋白表达水平升高，p62蛋白表达水平减少，SOD活性增强，MDA含量减少（P<0.05）；M组T₂—T₅时MWT和TWL均降低，T₅时脊髓LC3Ⅱ和Beclin-1蛋白表达水平减少，p62蛋白表达水平升高，SOD活性降低，MDA含量增加（P<0.05）。与M组比较，HM组T₂—T₅时MWT和TWL均升高，T₅时脊髓LC3Ⅱ和Beclin-1蛋白表达水平升高，p62蛋白表达水平减少，SOD活性增高，MDA含量减少（P<0.05）。结论富氢液可减轻大鼠神经病理性痛，抑制脊髓氧化应激，其机制可能与增加自噬有关。

关键词: 氢, 神经痛, 自噬, 脊髓, 氧化性应激

Abstract:

Objective To evaluate the role of autophagy in the treatment of neuropathic pain (NP) with hydrogen-rich saline. Methods Forty adult male Sprague-Dawley rats with successful intubation were randomly divided into 5 groups (n=8) using a random number table: the sham operation group (group S), the neuropathic pain group (group C), the hydrogen-rich saline group (group H), the autophagy inhibitor group (group M) and the hydrogen-rich saline + autophagy inhibitor group (group HM). There were 8 rats in each group. The NP model was established by chronic constriction of the sciatic nerve (CCI) in rats. The autophagy inhibitor 3-methyladenine (3-MA) was intraperitoneally injected with 30μg/kg in the group M and the group HM. The hydrogen-rich saline (0.6 mmol/L) was intraperitoneally injected with 10 mL/kg in the group H and the group HM. The other groups were intraperitoneally injected with the same amount of normal saline twice a day for 7 consecutive days. Paw withdrawal threshold to mechanical stimulation (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured at 1 day before and 1, 3, 5, 7 and 14 days after modeling (T₀-T₅). After the last measurement of pain threshold, the L4-L6 segment of spinal cord was removed for determination of the expression of autophagy-related proteins microtubule-associated protein light chain 3 (LC3) Ⅱ, Beclin-1 and p62 proteins by Western blot assay. The expression levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in spinal cord tissue were detected. Results Compared with the group S, MWT and TWL were decreased in the group C at T_2-5, the expression levels of LC3 Ⅱ, Beclin-1 and p62 were increased, SOD activity was decreased, and MDA content was increased at T₅(P<0.05). Compared with the group C, MWT and TWL were increased in the group H at T_2-5, LC3 Ⅱ and Beclin-1 protein expression levels were increased, p62 protein expression levels were decreased, SOD activity was increased, and MDA content was decreased at T₅ (P<0.05). MWT and TWL were decreased in the group M at T_2-5, LC3 Ⅱ and Beclin-1 protein expression levels were decreased, p62 protein expression levels were increased, SOD activity was decreased, and MDA content was increased at T₅(P<0.05). Compared with the group M, MWT and TWL were increased in the group HM at T_2-5, LC3 Ⅱ and Beclin-1 protein expression levels were increased, p62 protein expression levels were decreased, SOD activity was increased, and MDA content was decreased at T₅ (P<0.05). Conclusion Hydrogen-rich saline can alleviate neuropathic pain and inhibit oxidative stress in spinal cord in rats, and the mechanism may be related to the increase of autophagy.

Key words: hydrogen, neuralgia, autophagy, spinal cord, oxidative

中图分类号:

R614

图/表 5

参考文献 17

[1]	张昆龙, 薛白洁, 肖玮, 等. 重复经颅磁刺激对神经病理性疼痛患者疼痛和情绪的影响[J]. 中国现代神经疾病杂志, 2022, 22(11):940-947.
	ZHANG K L, XUE B J, XIAO W, et al. Effects of repetitive transcranial magnetic stimulation on pain and emotion of patients with neuropathic pain[J]. Chin J Contemp Neurol Neurosurg, 2022, 22(11):940-947. doi:10.3969/j.issn.1672-6731.2022.11.005.
[2]	CHEN H, ZHOU C, XIE K, et al. Hydrogen-rich saline alleviated the hyperpathia and microglia activation via autophagy mediated inflammasome inactivation in neuropathic pain rats[J]. Neuroscience, 2019, 421:17-30. doi:10.1016/j. neuroscience.2019.10.046.
[3]	CHEN Q, CHEN P, ZHOU S, et al. Hydrogen-rich saline attenuated neuropathic pain by reducing oxidative stress[J]. Can J Neurol Sci, 2013, 40(6):857-863. doi:10.1155/2019/8685954.
[4]	FENG T, YIN Q, WENG Z L, et al. Rapamycin ameliorates neuropathic pain by activating autophagy and inhibiting interleukin-1β in the rat spinal cord[J]. J Huazhong Univ Sci Technolog Med Sci, 2014, 34(6):830-837. doi:10.1007/s11596-014-1361-6.
[5]	BENNETT G J, XIE Y K. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man[J]. Pain, 1988, 33(1):87-107. doi:10.1016/0304-3959(88)90209-6.
[6]	ZHOU L, WANG X, XUE W, et al. Beneficial effects of hydrogen-rich saline against spinal cord ischemia-reperfusion injury in rabbits[J]. Brain Res, 2013, 1517:150-160. doi:10.1016/j.brainres.2013.04.007.
[7]	SPRINGER M Z, MACLEOD K F. Mitophagy:mechanisms and role in human disease[J]. J Pathol, 2016, 240(3):253-255. doi:10.1002/path.4774.
[8]	GHOSH A, TYSON T, GEORGE S, et al. Mitochondrial pyruvate carrier regulates autophagy,inflammation,and neurodegeneration in experimental models of Parkinson's disease[J]. Sci Transl Med, 2016, 8(368):368ra174. doi:10.1126/scitranslmed.aag2210.
[9]	ZHANG Z, SUN X, WANG K, et al. Hydrogen-saturated saline mediated neuroprotection through autophagy via PI3K/AKT/mTOR pathway in early and medium stages of rotenone-induced Parkinson's disease rats[J]. Brain Res Bull, 2021, 172:1-13. doi:10.1016/j.brainresbull.2021.04.003.
[10]	BARTH S, GLICK D, MACLEOD K F. Autophagy:assays and artifacts[J]. J Pathol, 2010, 221(2):117-124. doi:10.1002/path.2694.
[11]	LEE H S, DANIELS B H, SALAS E, et al. Clinical utility of LC3 and p62 immunohistochemistry in diagnosis of drug-induced autophagic vacuolar myopathies:a case-control study[J]. PLoS One, 2012, 7(4):e36221. doi:10.1371/journal.pone.0036221.
[12]	KUBISCH J, TÜREI D, FÖLDVÁRI-NAGY L, et al. Complex regulation of autophagy in cancer-integrated approaches to discover the networks that hold a double-edged sword[J]. Semin Cancer Biol, 2013, 23(4):252-261. doi:10.1016/j.semcancer.2013.06.009.
[13]	LI J, TIAN M, HUA T, et al. Combination of autophagy and NFE2L2/NRF₂ activation as a treatment approach for neuropathic pain[J]. Autophagy, 2021, 17(12):4062-4082. doi:10.1080/15548627.2021.1900498.
[14]	BUENDIA I, MICHALSKA P, NAVARRO E, et al. Nrf2-ARE pathway:an emerging target against oxidative stress and neuroinflammation in neurodegenerative diseases[J]. Pharmacol Ther, 2016, 157:84-104. doi:10.1016/j.pharmthera.2015.11.003.
[15]	BAXTER P S, BELL K F S, HASEL P, et al. Synaptic NMDA receptor activity is coupled to the transcriptional control of the glutathione system[J]. Nat Commun, 2015, 6:6761. doi:10.1038/ncomms7761.
[16]	MA W, MAO J, YANG X, et al. A single-atom Fe-N₄ catalytic site mimicking bifunctional antioxidative enzymes for oxidative stress cytoprotection[J]. Chem Commun (Camb), 2018, 55(2):159-162. doi:10.1039/c8cc08116f.
[17]	CERDÁ-BERNAD D, VALERO-CASES E, PASTOR J J, et al. Saffron bioactives crocin,crocetin and safranal:effect on oxidative stress and mechanisms of action[J]. Crit Rev Food Sci Nutr, 2022, 62(12):3232-3249. doi:10.1080/10408398.2020.1864279.

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	17.4±1.3	16.2±1.7	16.0±1.6	17.4±0.7	16.2±0.8	17.1±0.8	1.925
C组	16.6±1.0	15.5±1.2	12.4±1.2^ABa	9.6±1.8^ABCa	7.8±1.1^ABCDa	7.4±1.0^ABCDa	101.919^＊
H组	17.3±0.9	16.2±1.8	14.3±1.5^ABb	12.1±1.2^ABCb	11.0±1.2^ABCb	11.4±1.5^ABCb	42.582^＊
M组	16.7±1.1	14.8±1.4^A	10.5±1.6^ABb	7.9±1.1^ABCb	6.1±0.7^ABCDb	5.9±0.7^ABCDb	128.416^＊
HM组	17.1±1.2	15.5±0.8^A	12.1±1.4^ABc	9.9±1.5^ABCc	7.9±1.2^ABCDc	7.9±1.1^ABCDc	99.278^＊
F	0.936	1.344	16.697^＊	61.703^＊	117.144^＊	144.049^＊

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	17.4±1.3	16.2±1.7	16.0±1.6	17.4±0.7	16.2±0.8	17.1±0.8	1.925
C组	16.6±1.0	15.5±1.2	12.4±1.2^ABa	9.6±1.8^ABCa	7.8±1.1^ABCDa	7.4±1.0^ABCDa	101.919^＊
H组	17.3±0.9	16.2±1.8	14.3±1.5^ABb	12.1±1.2^ABCb	11.0±1.2^ABCb	11.4±1.5^ABCb	42.582^＊
M组	16.7±1.1	14.8±1.4^A	10.5±1.6^ABb	7.9±1.1^ABCb	6.1±0.7^ABCDb	5.9±0.7^ABCDb	128.416^＊
HM组	17.1±1.2	15.5±0.8^A	12.1±1.4^ABc	9.9±1.5^ABCc	7.9±1.2^ABCDc	7.9±1.1^ABCDc	99.278^＊
F	0.936	1.344	16.697^＊	61.703^＊	117.144^＊	144.049^＊

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	14.6±1.0	13.7±0.8	14.8±1.3	14.4±1.7	15.2±1.6	14.6±1.4	1.709
C组	14.5±1.3	13.1±1.7^A	10.2±1.6^ABa	7.7±1.0^ABCa	6.2±1.1^ABCDa	6.6±1.0^ABCa	74.429^＊
H组	15.2±1.3	13.5±1.1^A	12.4±1.2^ABb	10.5±1.5^ABCb	9.6±1.1^ABCb	10.0±1.1^ABCb	28.789^＊
M组	14.3±1.3	12.5±1.0^A	8.6±1.3^ABb	6.0±0.9^ABCb	4.9±0.8^ABCDb	5.3±0.8^ABCb	96.627^＊
HM组	14.5±1.3	13.1±1.5^A	10.5±1.4^ABc	8.2±1.2^ABCc	6.5±1.0^ABCDc	6.6±0.9^ABCDc	71.564^＊
F	0.630	1.136	24.649^＊	51.482^＊	103.909^＊	105.028^＊

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	14.6±1.0	13.7±0.8	14.8±1.3	14.4±1.7	15.2±1.6	14.6±1.4	1.709
C组	14.5±1.3	13.1±1.7^A	10.2±1.6^ABa	7.7±1.0^ABCa	6.2±1.1^ABCDa	6.6±1.0^ABCa	74.429^＊
H组	15.2±1.3	13.5±1.1^A	12.4±1.2^ABb	10.5±1.5^ABCb	9.6±1.1^ABCb	10.0±1.1^ABCb	28.789^＊
M组	14.3±1.3	12.5±1.0^A	8.6±1.3^ABb	6.0±0.9^ABCb	4.9±0.8^ABCDb	5.3±0.8^ABCb	96.627^＊
HM组	14.5±1.3	13.1±1.5^A	10.5±1.4^ABc	8.2±1.2^ABCc	6.5±1.0^ABCDc	6.6±0.9^ABCDc	71.564^＊
F	0.630	1.136	24.649^＊	51.482^＊	103.909^＊	105.028^＊

组别	LC3Ⅱ	Beclin-1	p62
S组	0.115±0.013	0.150±0.005	0.051±0.005
C组	1.510±0.012^a	0.916±0.029^a	0.286±0.009^a
H组	1.919±0.062^b	1.127±0.014^b	0.140±0.006^b
M组	0.289±0.031^b	0.215±0.010^b	0.401±0.010^b
HM组	1.068±0.735^c	0.908±0.012^c	0.287±0.009^c
F	4 722.678^＊	6 088.196^＊	2 362.972^＊

富氢液通过增加自噬治疗大鼠神经病理性疼痛

Hydrogen-rich saline treated neuropathic pain in rats by increasing autophagy

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组别	SOD/（U/mg）	MDA/（μmol/g）
S组	173.0±6.6	1.2±0.2
C组	148.3±14.0^a	4.3±0.9^a
H组	160.7±11.1^b	2.6±0.9^b
M组	75.1±13.4^b	14.2±1.2^b
HM组	147.4±13.1^c	4.4±0.9^c
F	81.815^＊	259.985^＊

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