电针调控PPARγ-CD36信号通路降低Hba-a1、Hbb-bt表达减轻脊髓损伤后细胞凋亡的研究  

Electroacupuncture reduces the expression of Hba-a1 and Hbb-bt and alleviates cell apoptosis after spinal cord injury by regulating the PPARγ-CD36 signaling pathway

作　　者：李明娇 唐成林[1] 杨祝歆 赵鸿娣 王嘉培 邢珂晗 黄思琴[1] Li Mingjiao;Tang Chenglin;Yang Zhuxin;Zhao Hongdi;Wang Jiapei;Xing Kehan;Huang Siqin(Chongqing Key Laboratory of Traditional Chinese Medicine Prevention and Treatment of Metabolic Disorders,College of Traditional Chinese Medicine,Chongqing Medical University)

机构地区：[1]重庆医科大学中医药学院中医防治代谢性疾病重庆市重点实验室,重庆400016

出　　处：《重庆医科大学学报》2025年第3期311-321,共11页Journal of Chongqing Medical University

基　　金：国家青年科学基金资助项目(编号:81403466);重庆市科委资助项目(编号:cstc2017jcyjAX0363);重庆市基础研究与前沿探索资助项目(编号:cstc2018jcyjAX0036);2020年重庆市科委卫计委资助项目(编号:2021ZY023890);2021年重庆市科技局资助项目(编号:cstc2021jcyjmsxmX0203)。

摘　　要：目的:本研究旨在建立脊髓损伤小鼠模型探讨电针(electroacupuncture,EA)干预小鼠急性脊髓损伤(spinal cord injury,SCI)后细胞凋亡的作用和机制。方法:采用雌性C 57 BL/6小鼠进行SCI造模,造模成功后使用随机法分为:脊髓损伤组(SCI组)、Electroacupuncture组(EA组)、Rosiglitazone组(R组),另设立假手术组(Sham组)。造模成功后对EA组小鼠取双侧“夹脊穴”和“足三里穴”进行电针,每日1次,连续治疗14 d;R组腹腔注射过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptor γ,PPARγ)激动剂:Rosiglitazone(5 mg/kg),Sham及SCI组不予特殊干预。采用小鼠后肢运动功能评分系统(basso mouse scale,BMS)评分评价运动功能,HE染色法、免疫荧光法检测脊髓损伤后的病理改变,转录组测序(RNA sequencing,RNA-Seq)、定量蛋白质组学(tandem mass tag/isobaric tags for relative and absolute quantification,TMT/iTRAQ)技术检测电针作用的靶点及机制,Western blot法检测PPARγ、分化簇36(cluster of differentiation 36,CD36)、血红蛋白亚基α1(hemoglobin al⁃pha,adult chain 1,Hba-a1)、血红蛋白β亚基(hemoglobin,beta adult t chain,Hbb-bt)、胱氨酸蛋白酶-3(Caspase-3,CASP3)蛋白表达水平。结果:与SCI组相比,EA组和R组的后肢运动功能、组织结构和残存细胞数量均有改善。EA组和R组Caspase-3表达降低,PPARγ和CD36表达增加,EA组Hba-a1和Hbb-bt表达降低。此外,RNA-Seq、TMT/iTRAQ技术结果分析、韦恩分析及双组学联合分析确定了Hba-a1和Hbb-bt为电针作用的靶基因。KEGG通路富集分析表明电针被证明对PPAR信号通路有明显影响。结论:电针可以通过调控PPARγ-CD36信号通路促进SCI后Hba-a1、Hbb-bt的清除,降低Caspase-3表达水平,减少细胞凋亡的发生,促进脊髓神经功能的恢复。Objective:To establish a mouse model of spinal cord injury(SCI),and to investigate the effect of electroacupuncture(EA)intervention on cell apoptosis after acute SCI and its mechanism.Methods:Female C57BL/6 mice were used to establish a model of SCI,and after successful modeling,the mice were randomly divided into SCI group,EA group,and Rosiglitazone group(R group);a sham-operation group(Sham group)was also established.After successful modeling,the mice in the EA group were given EA at bilateral Jiaji points and Zusanli once a day for 14 days,those in the R group were given intraperitoneal injection of the PPARγagonist Rosiglitazone(5 mg/kg),and those in the Sham group and the SCI group were not given any specific treatment.BMS score was used to assess motor function;HE staining and immunofluorescence assay were used to observe pathological changes after SCI;RNA-Seq and TMT/iTRAQ techniques were used to identify the targets and mechanisms of EA;Western blot was used to measure the protein expression levels of PPARγ,CD36,Hba-a1,Hbb-bt,and caspase-3.Results:Compared with the SCI group,the EA group and the R group had improvements in hindlimb motor function,tissue structure,and the number of surviving cells.The EA group and the R group had a significant reduction in the expression of caspase-3 and significant increases in the expression of PPARγ and CD36,and the EA group had significant reductions in the expression of Hba-a1 and Hbb-bt.In addition,RNA-Seq and TMT/iTRAQ techniques,significant analysis,Venn analysis,and dual-omics analysis identified Hba-a1 and Hbb-bt as the target genes of EA.The KEGG pathway enrichment analysis showed that EA had a significant effect on the PPAR signaling pathway.Conclusion:By regulating the PPARγ-CD36 signaling pathway,EA can promote the clearance of Hbaa1 and Hbb-bt after SCI,reduce the expression level of caspase-3,alleviate cell apoptosis,and facilitate the recovery of spinal cord nerve function.

关 键 词：电针 脊髓损伤 凋亡 转录组测序 定量蛋白质组学 

分 类 号：R651.2[医药卫生—外科学]