Effects of tetramethylpyrazine on analgesia and neuronal damage in migraine rats by regulating SIRT1/AMPK/PGC1α signaling pathway

doi:10.11958/20220873

Abstract

Abstract:

Objective To explore the effects of tetramethylpyrazine (TMP) on analgesia and neuronal injury protection in migraine rats by regulating silent mating type information regulation 2 homolog 1 (SIRT1)/AMP activated protein kinase (AMPK)/peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) signaling pathway. Methods The migraine rat model was established by nitroglycerin induction. After successful modeling, rats were randomly divided into the model (M) group, the TMP low dose (TMP-L) group (50 mg/kg), the TMP medium dose (TMP-M) group (100 mg/kg), the TMP high dose (TMP-M) group (200 mg/kg) and the TMP (200 mg/kg) + SIRT1 inhibitor (EX527, 5 mg/kg) group, 10 rats in each group. Another 10 rats were regarded as the normal control (NC) group. Rats were continuously gavaged for 2 weeks. Twenty-four hours after the end of the administration, the times of scratching head scratching and cage climbing of rats within 30 minutes were recorded in each group, and the behavioral score was carried out. The pain threshold for mechanical stimulation and thermal stimulation were determined. ELISA method was applied to measure serum levels of nitric oxide (NO), interleukin-6 (IL-6), interleukin-1β (IL-1β), and 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA) in brain tissue. TUNEL staining was applied to observe neuronal apoptosis in brain tissue. Western blot assay was applied to measure the protein expression levels of SIRT1, AMPK, p-AMPK and PGC1α protein in brain tissue. Results Compared with the NC group, the behavioral score, serum levels of NO, IL-6 and IL-1β, and neuron apoptosis rate were significantly increased in the M group (P<0.05). The pain threshold of mechanical stimulation was significantly reduced, and the latency of thermal stimulation was significantly shortened (P<0.05). The levels of 5-HT, NE and DA in brain tissue, the ratio of p-AMPK/AMPK, and the protein expressions of SIRT1 and PGC1α were significantly decreased (P<0.05). Compared with the M group, the behavioral score, the serum levels of NO, IL-6 and IL-1β, and neuron apoptosis rate were significantly decreased in the TMP groups (P<0.05). The pain threshold of mechanical stimulation was significantly increased, and the latency of thermal stimulation was significantly prolonged (P<0.05). The levels of 5-HT, NE and DA in brain tissue, the ratio of p-AMPK/AMPK, and the protein expressions of SIRT1 and PGC1α were significantly increased (P<0.05). Compared with the TMP-H group, TMP+EX527 group showed that it significantly reversed the effect of TMP on migraine rats. Conclusion TMP may improve neuronal damage by regulating the expression of SIRT1/AMPK/PGC1α signaling pathway and exert analgesic effect on migraine rats.

Key words: migraine, analgesia, neurons, AMP-activated protein kinases, peroxisome proliferator-activated receptor gamma coactivator 1α, nitric oxide, interleukin-6, interleukin-1beta, tetramethylpyrazine, silent mating type information regulation 2 homolog 1

CLC Number:

R285.5

Figures/Tables 7

References 22

[1]	SPECK R M, COLLINS E M, LOMBARD L, et al. A qualitative study to assess the content validity of the 24-hour migraine quality of life questionnaire in patients with migraine[J]. Headache, 2020, 60(9):1982-1994. doi:10.1111/head.13915.
[2]	BARRA M, DAHL F A, VETVIK K G, et al. A Markov chain method for counting and modelling migraine attacks[J]. Sci Rep, 2020, 10(1):3631. doi:10.1038/s41598-020-60505-5.
[3]	HIRATA K, UEDA K, KOMORI M, et al. Unmet needs in Japanese patients who report insufficient efficacy with trip-tans for acute treatment of migraine:Retrospective analysis of real-world data[J]. Pain Ther, 2021, 10(1):415-432. doi:10.1007/s40122-020-00223-y.
[4]	MENG Z, CHEN H, MENG S. The roles of tetramethylpyrazine during neurodegenerative disease[J]. Neurotox Res, 2021, 39(5):1665-1677. doi:10.1007/s12640-021-00398-y.
[5]	LI H, BAI F, CONG C, et al. Effects of ligustrazine on the expression of neurotransmitters in the trigeminal ganglion of a rat migraine model[J]. Ann Transl Med, 2021, 9(16):1318-1328. doi:10.21037/atm-21-3423.
[6]	王欢, 王淑敏, 陈长宝, 等. 黄绿卷毛菇对硝酸甘油诱导偏头痛大鼠的镇痛、抗炎作用[J]. 菌物学报, 2020, 39(5):917-922.
	WANG H, WANG S M, CHEN C B, et al. Anti-inflammatory and analgetic effects of the aqueous extract of Floccularia luteovirens on NTG-induced migraine in rats[J]. Mycosystema, 2020, 39(5):917-922. doi:10.13346/j.mycosystema.190384.
[7]	ZHENG Y, LIU T, WANG Z, et al. Low molecular weight fucoidan attenuates liver injury via SIRT1/AMPK/PGC1α axis in db/db mice[J]. Int J Biol Macromol, 2018, 112:929-936. doi:10.1016/j.ijbiomac.2018.02.072.
[8]	刘洁, 李利民, 宁楠, 等. 半夏泻心汤介导的cAMP/PKA信号通路对偏头痛模型大鼠c-fos/c-jun基因的调控作用[J]. 中药药理与临床, 2019, 35(3):37-40.
	LIU J, LI L M, NING N, et al. The regulation of c-fos/c-jun by (Banxiaxiexintang) BXT mediated CAMP/PKA signaling pathway in migraine rat model[J]. Pharmacology and Clinics of Chinese Materia Medica, 2019, 35(3):37-40. doi:10.13412/j.cnki.zyyl.2019.03.008.
[9]	李莉, 张琼果, 潘巧虹, 等. 基于胞外信号调节激酶/胞浆型磷脂酶A2信号通路分析川穹嗪对膜性肾病大鼠肾保护作用[J]. 中国临床药理学杂志, 2020, 36(12):1665-1668.
	LI L, ZHANG Q G, PAN Q H, et al. Study on renal protection effect of tetramethylpyrazine on membranous nephropathy rats based on extracellular signal-regulated kinase/cytosolic phospholipase A2 signaling pathway[J]. Chin J Clin Pharmacol, 2020, 36(12):1665-1668. doi:10.13699/j.cnki.1001-6821.2020.12.017.
[10]	肖志博, 谢海, 金辉, 等. 丙泊酚基于SIRT1/Foxo1信号缓解脑缺血再灌注造成血脑屏障损伤[J]. 微循环学杂志, 2022, 32(1):12-18.
	XIAO Z B, XIE H, JIN H, et al. Protection of propofol on blood-brain barrier injury induced by ischemia/reperfusion based on SIRT1/Foxo1 signal[J]. Chinese Journal of Microcirculation, 2022, 32(1):12-18. doi:10.3969/j.issn.1005-1740.2022.01.003.
[11]	王萌萌, 于晓华, 耿炜, 等. 疏肝调神针法对偏头痛大鼠行为学及血中相关神经肽、炎性物质表达的影响[J]. 针刺研究, 2018, 43(6):375-379.
	WANG M M, YU X H, GENG W, et al. Effect of manual acupuncture preconditioning on behavior and contents of serum CGRP,SP,IL-1β and TNF-α levels in migraine rats[J]. Acupuncture Research, 2018, 43(6):375-379. doi:10.13702/j.1000-0607.170415.
[12]	姚庆萍, 邵淑娟. 偏头痛中医药治疗研究近况[J]. 内蒙古中医药, 2017, 36(11):97-99.
	YAO Q P, SHAO S J. Recent research on TCM treatment of migraine[J]. Inner Mongolia Journal of Traditional Chinese Medicine, 2017, 36(11):97-99. doi:10.16040/j.cnki.cn15-1101.2017.11.096.
[13]	EDVINSSON L, HAANES K A, WARFVINGE K, et al. CGRP as the target of new migraine therapies - successful translation from bench to clinic[J]. Nat Rev Neurol, 2018, 14(6):338-350. doi:10.1038/s41582-018-0003-1.
[14]	ZHANG C, SHEN M, TENG F, et al. Ultrasound-enhanced protective effect of tetramethylpyrazine via the ROS/HIF-1A signaling pathway in an in vitro cerebral Ischemia/reperfusion injury model[J]. Ultrasound Med Biol, 2018, 44(8):1786-1798. doi:10.1016/j.ultrasmedbio.2018.04.005.
[15]	邢娟, 范崇桂, 沈雷, 等. 基于PKC/P2X3通路探讨正天丸对偏头痛大鼠的保护作用[J]. 上海中医药杂志, 2020, 54(5):87-92.
	XING J, FAN C G, SHEN L, et al. Protective effect of Zhengtian pill on migraine rats based on PKC/P2X3 pathway[J]. Shanghai Journal of Traditional Chinese Medicine, 2020, 54(5):87-92. doi:10.16305/j.1007-1334.2020.05.008.
[16]	HOU M, TANG Q, XUE Q, et al. Pharmacodynamic action and mechanism of Du Liang soft capsule,a traditional Chinese medicine capsule,on treating nitroglycerin-induced migraine[J]. J Ethnopharmacol, 2017, 195:231-237. doi:10.1016/j.jep.2016.11.025.
[17]	刘宇光, 王欢, 王淑敏, 等. 药用真菌中麦角甾醇对偏头痛大鼠的作用机制[J]. 食品工业科技, 2021, 42(15):327-331.
	LIU Y G, WANG H, WANG S M, et al. Effects of ergosterol from medicinal fungi on migraine rats[J]. Science and Technology of Food Industry, 2021, 42(15):327-331. doi:10.13386/j.issn1002-0306.2020090049.
[18]	WEN W, CHEN H, FU K, et al. Fructus Viticis methanolic extract attenuates trigeminal hyperalgesia in migraine by regulating injury signal transmission[J]. Exp Ther Med, 2020, 19(1):85-94. doi:10.3892/etm.2019.8201.
[19]	SUN X, ZHU F, ZHOU J, et al. Anti-migraine and anti-depression activities of Tianshu capsule by mediating Monoamine oxidase[J]. Biomed Pharmacother, 2018, 100:275-281. doi:10.1016/j.biopha.2018.01.171.
[20]	ZHANG Y M, QU X Y, TAO L N, et al. XingNaoJing injection ameliorates cerebral ischaemia/reperfusion injury via SIRT1-mediated inflammatory response inhibition[J]. Pharm Biol, 2020, 58(1):16-24. doi:10.1080/13880209.2019.1698619.
[21]	YUN Y C, JEONG S G, KIM S H, et al. Reduced sirtuin 1/adenosine monophosphate-activated protein kinase in amyotrophic lateral sclerosis patient-derived mesenchymal stem cells can be restored by resveratrol[J]. J Tissue Eng Regen Med, 2019, 13(1):110-115. doi:10.1002/term.2776.
[22]	刘垚君, 张玉琴, 方雅玲, 等. 黄精多糖调控SIRT1/AMPK/PGC-1α信号通路改善H₂O₂诱导的HT22细胞氧化损伤[J]. 中国现代应用药学, 2021, 38(16):1952-1957.
	LIU Y J, ZHANG Y Q, FANG Y L, et al. Regulation of SIRT1/AMPK/PGC-1α signaling pathway by polygonatum polysaccharide improves H₂O₂-induced oxidative damage in HT22 cells[J]. Chin J Mod Appl Pharm, 2021, 38(16):1952-1957. doi:10.13748/j.cnki.issn1007-7693.2021.16.005.

组别	0~30 min	60~90 min	120~150 min
NC组	11.71±1.52	12.42±1.54	11.33±1.10
M组	44.85±3.96^a	33.49±2.62^a	22.45±1.86^a
TMP-L组	37.05±2.97^b	26.39±2.43^b	16.85±1.72^b
TMP-M组	25.12±2.18^bc	21.21±1.58^bc	13.23±1.16^bc
TMP-H组	20.02±1.94^bcd	18.32±1.34^bcd	12.42±1.79^bc
TMP+EX527组	29.97±2.01^e	27.86±1.13^e	18.12±1.24^e
F	215.487^**	163.238^**	77.599^**

组别	0~30 min	60~90 min	120~150 min
NC组	11.71±1.52	12.42±1.54	11.33±1.10
M组	44.85±3.96^a	33.49±2.62^a	22.45±1.86^a
TMP-L组	37.05±2.97^b	26.39±2.43^b	16.85±1.72^b
TMP-M组	25.12±2.18^bc	21.21±1.58^bc	13.23±1.16^bc
TMP-H组	20.02±1.94^bcd	18.32±1.34^bcd	12.42±1.79^bc
TMP+EX527组	29.97±2.01^e	27.86±1.13^e	18.12±1.24^e
F	215.487^**	163.238^**	77.599^**

组别	机械性刺激痛阈（g）	热刺激潜伏期（s）
NC组	9.72±0.75	8.65±0.73
M组	5.79±0.83^a	4.12±0.36^a
TMP-L组	6.73±0.63^b	5.67±0.42^b
TMP-M组	7.67±0.55^bc	6.45±0.38^bc
TMP-H组	8.55±0.68^bc	7.25±0.73^bcd
TMP+EX527组	7.57±0.75^e	6.33±0.57^e
F	37.927^**	75.104^**

组别	机械性刺激痛阈（g）	热刺激潜伏期（s）
NC组	9.72±0.75	8.65±0.73
M组	5.79±0.83^a	4.12±0.36^a
TMP-L组	6.73±0.63^b	5.67±0.42^b
TMP-M组	7.67±0.55^bc	6.45±0.38^bc
TMP-H组	8.55±0.68^bc	7.25±0.73^bcd
TMP+EX527组	7.57±0.75^e	6.33±0.57^e
F	37.927^**	75.104^**

组别	NO（mmol/L）	IL-6（μg/L）	IL-1β（μg/L）
NC组	12.76±1.12	0.25±0.03	1.41±0.12
M组	34.51±2.97^a	3.69±0.32^a	5.14±0.48^a
TMP-L组	28.65±2.31^b	2.41±0.19^b	3.76±0.37^b
TMP-M组	23.87±1.73^bc	1.64±0.23^bc	2.97±0.26^bc
TMP-H组	17.52±1.33^bcd	0.72±0.05^bcd	1.92±0.24^bcd
TMP+EX527组	27.39±2.54^e	1.66±0.16^e	2.89±0.21^e
F	140.548^**	409.845^**	193.031^**