摘要
目的 探究川穹嗪(TMP)通过调控沉默信息调节因子2相关酶1(SIRT1)/腺苷酸活化蛋白激酶(AMPK)/过氧化物酶体增殖物激活受体γ共激活因子1α(PGC1α)信号通路对偏头痛大鼠发挥镇痛和神经元损伤的保护作用。方法 通过硝酸甘油诱导建立偏头痛大鼠模型,造模成功后随机分为模型(M)组、TMP低剂量(TMP-L)组(50 mg/kg)、TMP中剂量(TMP-M)组(100 mg/kg)、TMP高剂量(TMP-H)组(200 mg/kg)、TMP(200 mg/kg)+SIRT1抑制剂(EX527,5 mg/kg)组,每组10只;另取10只作为正常对照(NC)组。连续灌胃2周。给药结束24 h后,记录各组大鼠在连续30 min内出现挠头、爬笼的次数,进行行为学评分;测定机械性刺激及热刺激痛阈;酶联免疫吸附试验法检测血清中一氧化氮(NO)、白细胞介素(IL)-6、IL-1β含量和脑组织中5-羟色胺(5-HT)、去甲肾上腺素(NE)、多巴胺(DA)含量;TUNEL染色观察脑组织神经元凋亡情况;Western blot法检测脑组织中SIRT1、AMPK、p-AMPK、PGC1α蛋白表达。结果 与NC组比较,M组大鼠行为学评分,血清中NO、IL-6、IL-1β水平,神经元凋亡率升高(P<0.05);机械性刺激痛阈值降低,热刺激潜伏期缩短(P<0.05);脑组织中5-HT、NE、DA水平,p-AMPK/AMPK比值,SIRT1、PGC1α蛋白表达降低(P<0.05)。与M组比较,TMP各剂量组大鼠行为学评分,血清中NO、IL-6、IL-1β水平,神经元凋亡率降低(P<0.05);机械性刺激痛阈值升高,热刺激潜伏期延长(P<0.05);脑组织中5-HT、NE、DA水平,pAMPK/AMPK比值,SIRT1、PGC1α蛋白表达升高(P<0.05);与TMP-H组比较,TMP+EX527组可显著逆转TMP对偏头痛大鼠的作用。结论 TMP可能通过调节SIRT1/AMPK/PGC1α信号通路的表达,改善神经元损伤,发挥对偏头痛大鼠的镇痛作用。
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 receptorγcoactivator 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.Twentyfour 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 PGC1αprotein 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 weresignificantly decreased in the TMP groups (P<0.05). The pain threshold of mechanical stimulation was significantlyincreased, and the latency of thermal stimulation was significantly prolonged (P<0.05). The levels of 5-HT, NE and DA inbrain 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 onmigraine rats. Conclusion TMP may improve neuronal damage by regulating the expression of SIRT1/AMPK/PGC1αsignaling pathway and exert analgesic effect on migraine rats.
作者
胡滨
王大斌
郭茂
HU Bin;WANG Dabin;GUO Mao(Department of Pain,Luzhou People's Hospital,Luzhou 646000,China)
出处
《天津医药》
CAS
北大核心
2023年第4期382-387,共6页
Tianjin Medical Journal
