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

李明娇; 唐成林; 杨祝歆; 赵鸿娣; 王嘉培; 邢珂晗; 黄思琴

doi:10.13406/j.cnki.cyxb.003784

重庆医科大学学报 ›› 2025, Vol. 50 ›› Issue (03) : 311 -321. DOI: 10.13406/j.cnki.cyxb.003784

基础研究

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

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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

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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 alpha,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。

Abstract

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 PPARγ agonist 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 Hba-a1 and Hbb-bt after SCI，reduce the expression level of caspase-3，alleviate cell apoptosis，and facilitate the recovery of spinal cord nerve function.