【作者】 王列； 李记泉； 马帅； 于嘉祥； 马俊杰； 胡哲； 李格格； 陈怡然； 王鹰； 卞镝； 马铁明； 董宝强； 于本性； 王树东；

【Author】 WANG Lie;LI Ji-quan;MA Shuai;YU Jia-xiang;MA Jun-jie;HU Zhe;LI Ge-ge;CHEN Yi-ran;WANG Ying;BIAN Di;MA Tie-ming;DONG Bao-Qiang;YU Ben-xing;WANG Shu-dong;Liaoning University of Traditional Chinese Medicine;Key Laboratory of Acupuncture Moxibustion and Health Rehabilitation in Liaoning Province;Shenyang Seventh People’s Hospital;

【机构】 辽宁中医药大学； 辽宁省针灸养生康复重点实验室； 沈阳市第七人民医院；

【摘要】 目的筛查电针治疗肌筋膜疼痛综合征(MPS)模型大鼠引起信使RNA(m RNA)表达水平变化的基因。方法7周健康的无特定病原体(SPF)级斯泼累格·多雷(SD)大鼠,分别设置空白组、模型组、电针组,采用钝性打击结合离心运动方式复制MPS的模型,对电针组进行电针干预,取材前进行各组大鼠自发电活动频率,热缩足潜伏期检测,取材后首先通过Elisa法检测各组大鼠血清中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-px)、过氧化氢酶(CAT)、白介素-1β(IL-1 β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)值,然后通过苏木精-伊红染色法(HE)观察各组大鼠肌肉组织病理形态,最后以各组大鼠的脊髓组织为研究对象进行转录组学测序,并应用平行反应监视技术(PRM)对序列相似家族111成员A(FAM111A)、锌指和含BTB域16(ZBTB16)、蛋白磷酸酶1调节因子亚基3G(PPP1R3G)、围脂滴蛋白4(PLIN4)、蛋白磷酸酶1调节亚基3C(PPP1R3C)共5个蛋白进行定量。结果电针干预能抑制MPS模型大鼠的氧化应激及炎症水平,可引起MPS模型大鼠87个基因的m RNA表达水平发生变化,涉及到了钙离子、叉头框蛋白O1(Fox O1)等信号通路。电针干预能升高FAM111A的表达水平,降低ZBTB16、PPP1R3G、PLIN4、PPP1R3C的表达水平,差异表达的基因和富集显著性靠前的通路与对炎症、氧化应激的抑制和对能量代谢调控相关。结论实验所采取的技术手段合理,样本稳定,电针干预MPS有效;电针治疗MPS的具体机制与抑制炎症、氧化应激和调控能量代谢有关,最终修复了因MPS造成的病理损伤,转录组学差异基因所富集的通路及重要分子的机制直接或间接证实了上述结论。更多还原

【Abstract】 Screening for genes that cause changes in mRNA expression levels in rats with myofascial pain syndrome(MPS) treated with electroacupuncture.Method:Seven weeks of healthy SPF grade SD rats were divided into a blank group,a model group,and an electroacupuncture group.The MPS model was replicated using blunt impact combined with centrifugal movement.After electroacupuncture intervention,Before sampling,the frequency of self generating activity and the latent period of thermal contraction in each group of rats were detected.Before sampling,the frequency of self generating activity and the latent period of thermal contraction in each group of rats were detected.After sampling,the serum levels of SOD、MDA、GSH-px、CAT、IL-1 β、IL-6、TNF-α were detected by Elisa.Then,HE staining was used to observe the pathological morphology of muscle tissue in each group of rats.Finally,transcriptomic sequencing was performed on the spinal cord tissue of each group of rats,and PRM technology was used to quantify five proteins including FAM111A,ZBTB16,PPP1R3G,PLIN4,and PPP1R3C.Result,electroacupuncture intervention can inhibit oxidative stress and inflammation levels in MPS model rats,and can cause 87 genes changes in mRNA expression levels,including signaling pathways such as:Calcium ions and FoxO1.Electroacupuncture intervention can increase the expression level of FAM111A and reduce the expression levels of ZBTB16,PPP1R3G,PLIN4,and PPP1R3C,differentially expressed genes and pathways with higher enrichment significance are associated with inhibition of inflammation,oxidative stress,and regulation of energy metabolism.Conclusion:The technical means adopted in the experiment are reasonable,the sample is stable,and electroacupuncture intervention in MPS is effective;The specific mechanism of electroacupuncture for treating MPS is related to the inhibition of inflammation,oxidative stress,and regulation of energy metabolism,ultimately repairing the pathological damage caused by MPS.The pathways enriched by transcriptomic differentially expressed genes and the mechanisms of important molecules directly or indirectly confirm the above conclusion.更多还原