转运蛋白通过激活脊髓背角星形胶质细胞自噬缓解神经病理性疼痛  

Translocator protein alleviates neuropathic pain through activating the spinal cord dorsal horn astrocytes autophagy

作　　者：郝璨 马冰洁[1] 靳天 高楠 纪运 马柯[1] 刘晓明[1] HAO Can;MA Bingjie;JIN Tian;GAO Nan;JI Yun;MA Ke;LIU Xiaoming(Department of Pain Management,Xinhua Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200092,China)

机构地区：[1]上海交通大学医学院附属新华医院疼痛科,上海200092

出　　处：《中国疼痛医学杂志》2022年第1期21-29,共9页Chinese Journal of Pain Medicine

基　　金：国家自然科学基金(81771184)。

摘　　要：目的:探讨18kDa转运蛋白(translocator protein,TSPO)是否可以通过激活脊髓背角星形胶质细胞自噬来缓解神经病理性疼痛及其可能的机制。方法:实验分为两部分,第一部分将雄性SD大鼠按随机数字表法分为假手术组(Sham组)、对照组(SNL组)、TSPO受体激动剂Ro5-4864处理组(Ro组);第二部分将脊神经结扎模型(spinal nerve ligation,SNL)术后的雄性SD大鼠随机分为TSPO受体激动剂Ro5-4864处理组(Ro组)和自噬诱导剂Rapamycin处理组(Rap组)。采用von Frey纤维丝测量大鼠50%机械缩足反射阈值(mechanical withdrawal threshold,MWT),在术后第7天(D7)、14天(D14)取损伤侧脊髓样本,采用蛋白印迹技术检测自噬相关蛋白Beclin1、LC3、p62及通路蛋白PGC-1α,采用免疫荧光检测LC3在脊髓背角星形胶质细胞的表达。结果:第一部分实验结果:与Sham组相比,SNL组50%MWT从术后第1天开始明显下降,持续到观察结束,脊髓Beclin1、LC3、p62表达明显升高(P<0.01),脊髓PGC-1α表达明显降低,脊髓背角星形胶质细胞LC3表达信号增强;与SNL组相比,Ro组50%MWT在D7、D14明显升高,Ro组脊髓Beclin1、LC3表达明显升高(P<0.01),p62蛋白含量先升高后降低,PGC-1α持续升高,显著高于同时间SNL组(P<0.01)。第二部分实验结果:与Ro组相比,Rap组50%MWT在D7降低,Rap组脊髓D14 Beclin1及D7、D14 LC3差异有统计学意义(P<0.01)。结论:鞘内注射TSPO受体激动剂Ro5-4864能显著缓解神经病理性疼痛,其涉及机制之一可能为通过激活PGC-1α相关途径,而后激活脊髓背角星形胶质细胞的自噬。Objective:To study whether 18kDa translocator protein(TSPO)can alleviate neuropathic pain by activating astrocyte autophagy in the spinal dorsal horn and explore its possible mechanism.Methods:The experiment was divided into two parts.In the first part,male SD rats were randomly divided into 3 groups:The Sham group,the control group(SNL group),the Ro5-4864 group(Ro group).In the second part,male SD rats after spinal nerve ligation(SNL)surgery treatment were randomly divided into 2 groups:the Ro5-4864 group(Ro group)and the Rapamycin group(Rap group).von Frey hairs were used to measure the 50%mechanical withdrawal threshold(MWT).The protein expression of Beclin1,LC3,p62,and PGC-1αin the ipsilateral spinal cord was detected by western blot.The expression of LC3 in astrocytes of spinal dorsal horn was detected by immunofluorescence.Results:In the first part,compared to the sham group,the 50%MWT of the SNL group was significantly decreased,and the expression of Beclin1,LC3,and p62 were significantly increased(P<0.01),and the expression of PGC-1αwas significantly decreased.The expression of LC3 in astrocytes of spinal dorsal horn was increased in immunofluorescence intensity;Compared to the SNL group,the 50%MWT in the Ro group was increased significantly,and the expression of Beclin1,LC3,and PGC-1αin the Ro group were increased significantly(P<0.01).The expression of p62 in the Ro group was decreased compared to the SNL group(P<0.01).In the second part,compared to the Ro group,the 50%MWT of the Rap group was significantly decreased and the expression of Beclin1 and LC3 were significantly decreased in the Rap group(P<0.01).Conclusion:Intra-thecal injection of TSPO agonist Ro5-4864 can alleviate neuropathic pain.One possible mechanism is the activation of PGC-1αrelevant pathway and then activation of astrocytes autophagy in the spinal cord dorsal horn.

关 键 词：转运蛋白 神经病理性疼痛 星形胶质细胞 自噬 PGC-1Α 

分 类 号：R402[医药卫生—临床医学]