天麻素对神经病理性痛大鼠脊髓AMPK/TRPA1信号通路的影响
Effect of gastrogin on AMPK/TRPA1 signaling pathway in rats with neuropathic pain
孙雪 1王士雷 1卢衍羽 1赵洋1
作者信息
- 1. 青岛大学附属医院麻醉科,青岛 266555
- 折叠
摘要
目的 评价天麻素对神经病理性痛大鼠脊髓腺苷酸激活蛋白激酶(AMPK)/瞬时受体电位通道A1(TRPA1)信号通路的影响.方法 SPF级健康雄性SD大鼠36只,6～8周龄,体质量200～230 g,采用随机数字表法分为3组(n=12):假手术+生理盐水组(SHAM组)、神经病理性痛+生理盐水组(NP组)和神经病理性痛+天麻素组(GAS组).2％异氟烷麻醉下,采用坐骨神经慢性结扎损伤法制备大鼠神经病理性痛模型,SHAM组未结扎.造模后连续14 d,GAS组腹腔注射天麻素100 mg/kg,SHAM组和NP组注射等量生理盐水.分别于造模前1 d(T0)、造模后第1、3、5、7、10和14天(T1-6)时测定机械缩足反应阈(MWT)和热缩足潜伏期(TWL).在T4和T6时测完痛阈后,2％异氟烷麻醉处死大鼠取L4-6脊髓组织,采用qRT-PCR法检测TRPA1 mRNA表达,Western blot法检测脊髓TRPA1、AMPK和p-AMPK表达,免疫荧光染色法检测脊髓TRPA1表达;免疫组织化学染色法观察TNF-α、IL-1β和c-fos的表达.结果 与SHAM组相比,NP组T1-6时MWT和TWL降低,脊髓TRPA1 mRNA、TRPA1、TNF-α、IL-1β和c-fos表达上调,p-AMPK表达下调(P＜0.05),AMPK表达差异无统计学意义(P＞0.05);与 NP 组相比,GAS组T3.6时MWT和T2.6时TWL升高,脊髓TRPA1 mRNA、TRPA1、TNF-α、IL-1β和c-fos表达下调,p-AMPK表达上调(P＜0.05),AMPK表达差异无统计学意义(P＞0.05).结论 天麻素减轻大鼠神经病理性痛的机制可能与调控AMPK/TRPA1信号通路有关.
Abstract
Objective To evaluate the effect of gastrogin on AMP-activated protein kinase(AMPK)/transient receptor potential anchor protein 1(TRPA1)signaling pathway in rats with neuropathic pain.Methods Thirty-six SPF-grade healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 200-230 g,were divided into 3 groups(n=12 each)using a random number table method:sham operation+nor-mal saline group(SHAM group),neuropathic pain+normal saline group(NP group),and neuropathic pain+gastrogin group(GAS group).Neuropathic pain was induced by chronic constrictive injury to sciatic nerve under 2％ isoflurane anaesthesia.The sciatic nerve was only exposed but not ligated in SHAM group.Gastrogin 100 mg/kg was intraperitoneally injected for 14 consecutive days after developing the model in GAS group,while the equal volume of normal saline was given instead in SHAM and NP groups.The me-chanical paw withdrawal threshold(MWT)and thermal paw withdrawal latency(TWL)were measured at 1 day before developing the model(T0)and 1,3,5,7,10 and 14 days after developing the model(T1-6).The rats were anesthetized and sacrificed following the measurement of pain thresholds at T4 and T6.The lumbar segment(L4.6)of the spinal cord was removed for determination of TRPA1 mRNA expression(by quantitative real-time polymerase chain reaction),expression of TRPA1,AMPK and p-AMPK(by Western blot),expression of TRPA1(by immunofluorescence staining)and expression of tumor necrosis-alpha(TNF-α),interleukin-1 beta(IL-1β)and c-fos(by immunohistochemistry).Results Compared with SHAM group,MWT and TWL were significantly decreased at T1-6,the expression of TRPA1 mRNA,TR-PA1,TNF-α,IL-1β and c-fos was up-regulated,the expression of p-AMPK was down-regulated(P＜0.05),and no significant change was found in AMPK expression in NP group(P＞0.05).Compared with NP group,MWT at T3.6 and TWL at T2-6 were significantly increased,the expression of TRPA1 mRNA,TR-PA1,TNF-α,IL-1β and c-fos was down-regulated,and p-AMPK expression was up-regulated(P＜0.05),and no significant change was found in AMPK expression in GAS group(P＞0.05).Conclusions The mechanism by which gastrogin reduces neuropathic pain may be related to modulating the expression of the AMPK/TRPA1 signaling pathway in rats.
关键词
天麻/神经痛/TRPA1离子通道/AMP活化蛋白激酶类Key words
Gastrogin/Neuralgia/TRPA1 cation channel/AMP-activated protein kinases引用本文复制引用
基金项目
山东省医学会临床科研基金齐鲁专项(YXH2022ZX02095)
青岛大学附属医院临床医学+X科研项目(QDFY+X2023148)
出版年
2024
NETL
NSTL
维普
万方数据