石蒜碱靶向激活脊髓AMP活化蛋白激酶通路缓解大鼠关节炎痛的机制研究  

Lycorine alleviates arthritis pain in rats via AMPK-Bax/IL-1βsignaling

作　　者：胡引娣 陈春喜 谢敏 李岱[1] 王柏军[1] HU Yin-di;CHEN Chun-xi;XIE Min;LI Dai;WANG Bo-jun(School of Pharmacy,Xianning Medical College,Hubei University of Science and Technology,Xianning 437100,China;Xishui Hospital Affiliated to Hubei Institute of Science and Technology,Huanggang 438299,China)

机构地区：[1]湖北科技学院医学部药学院,湖北咸宁437100 [2]湖北科技学院附属浠水医院,湖北黄冈438802

出　　处：《中国病理生理杂志》2022年第12期2205-2212,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81901149);湖北省教育厅项目(No.B2019167);湖北科技学院校级团队项目(No.2020TD02)。

摘　　要：目的:研究石蒜碱缓解大鼠关节炎痛的作用及机制。方法:将40只雄性Sprague-Dawley大鼠随机分成对照组、模型组、石蒜碱给药组及AMP活化蛋白激酶(AMPK)激动剂AICAR给药组,每组10只;采用左后膝关节腔内注射完全弗氏佐剂构建大鼠关节炎模型,鞘内注射石蒜碱或AICAR进行给药,记录大鼠痛觉行为及运动能力变化;分子对接研究石蒜碱的作用靶点;免疫荧光和Western blot法检测各组动物脊髓组织中炎症相关蛋白表达的变化。结果:与对照组相比,完全弗氏佐剂注射14 d可造成大鼠自发缩足次数增加、机械痛阈值下降等运动能力受损情况(P<0.05),脊髓组织AMPK活性下调(P<0.05),同时星形胶质细胞标志物胶质细胞原纤维酸性蛋白(GFAP)、白细胞介素1β(IL-1β)、Bcl-2相关X蛋白(Bax)和cleaved caspase-3蛋白水平均显著升高(P<0.05)。与模型组相比,石蒜碱鞘内给药可显著减少大鼠自发缩足次数,增加机械痛阈值,延长转棒停留时间,增加运动距离(P<0.05),激活AMPK(P<0.05),降低GFAP、IL-1β、Bax和cleaved caspase-3蛋白水平(P<0.05)。结论:石蒜碱可靶向激活脊髓AMPK,抑制Bax/IL-1β炎症信号,从而缓解大鼠关节炎痛。AIM:To investigate the effect of lycorine on arthritis pain in rats and its mechanism.METHODS:Forty rats were randomly divided into 4 groups:control group,model group,lycorine group and AICAR[an activator of AMP-activated protein kinase(AMPK)]group,with 10 rats in each group.Rat model of arthritis was established by intraluminal injection of complete Freund’s adjuvant(CFA)in the left posterior knee joint.Lycorine and AICAR were intrathecally administered to the rats.Changes in pain behavior and motor performance of rats were detected.Molecular docking was used to study the target of lycorine.Immunofluorescence and Western blot were used to detect the expression of inflammation-related proteins in the spinal cord.RESULTS:Compared with control group,CFA injection induced the increase in the number of spontaneous flinches,the decrease in mechanical pain threshold and the impairment of the motor performance(P<0.05).CFA injection also down-regulated the activity of spinal AMPK and increased spinal expression levels of astrocytic marker glial fibrillary acidic protein(GFAP),interleukin-1β(IL-1β),Bcl-2-associated X protein(Bax)and cleaved caspase-3(P<0.05).Compared with model group,after intrathecal administration of lycorine,the number of spontaneous flinches was reduced,while the threshold of mechanical pain,and the latency and motor distance in rotarod test were increased(P<0.05).In addition,AMPK activity was up-regulated,and the protein levels of GFAP,IL-1β,Bax and cleaved caspase-3 were decreased(P<0.05).CONCLUSION:Targeting AMPK activation by lycorine inhibits the Bax/IL-1βinflammatory signaling in spinal cord and relieves arthritis pain in rats.

关 键 词：石蒜碱 骨关节炎 脊髓炎症 AMP活化蛋白激酶 白细胞介素1Β 

分 类 号：R593.22[医药卫生—内科学] R363.2[医药卫生—临床医学]