摘要
目的:观察可视化针刀对股骨头坏死(osteonecrosis of the femoral head,ONFH)疼痛触发点模型比格犬的影响,并基于转录组学探讨作用机制。方法:将24只比格犬随机分为对照组、模型组、可视化针刀组、传统针刀组,每组6只。除对照组外,其余比格犬建立ONFH疼痛触发点模型。可视化针刀组和传统针刀组各治疗3周,每周1次。采用ELISA法检测血清P物质(substance P,SP)、5-羟色胺(5-hydroxytryptamine,5-HT)、神经肽Y(neuropeptide Y,NPY)、降钙素基因相关肽1(calcitonon gene related peptide 1,CGRP1)、一氧化氮(nitric oxide,NO)含量,HE染色观察髋关节周围组织病理形态。采用高通量测序技术(RNA-seq)检测差异表达基因(differentially expressed genes,DEGs),并进行基因本体(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析。通过STRING数据库和Cytoscape软件构建ONFH神经递质代谢网络,相关基因与DEGs取交集,从而筛选核心靶点。结果:髋部CT显示,对照组比格犬髋关节未见异常;模型组比格犬髋关节在位,股骨头可见低密度区。髋关节MRI显示,对照组比格犬髋关节存在少量积液;模型组比格犬髋关节存在积液,股骨头可见双线征和混杂信号,伴骨髓水肿。治疗前,与对照组比较,模型组、传统针刀组及可视化针刀组比格犬血清SP、5-HT、NPY含量升高,NO、CGRP含量降低(P<0.05)。与治疗前比较,治疗21 d后传统针刀组、可视化针刀组比格犬血清SP、5-HT、NPY含量减少,NO、CGRP含量增加(P<0.05)。治疗21 d后,与传统针刀组比较,可视化针刀组比格犬血清SP、5-HT、NPY含量减少,NO、CGRP含量增加(P<0.05)。HE染色显示,对照组:细胞质染色均匀,细胞核排列整齐;模型组:细胞核排列紊乱,肌纤维松散伴断裂,肌横纹模糊,局部结缔组织增生;传统针刀组:细胞核排列稍紊乱,局部肌横纹模糊,结缔组织稍有增生;可视化针刀组:细胞核排列较整齐,肌纤维排列紧密。转录组学结果显示,与对照组比较,模型组下调基因929个;与模型组比较,可视化针刀组上调基因3265个,两者的交集基因685个。与对照组比较,模型组上调基因272个;与模型组比较,可视化针刀组下调基因748个,两者的交集基因142个。GO功能分析主要涉及对抗原加工和呈递的正向调节、磷脂酰肌醇3-激酶复合物IB类、鞘磷脂磷酸二酯酶活性等。KEGG通路主要包括破骨细胞分化、类风湿关节炎、吞噬细胞等。核心靶点包括乙酰胆碱酯酶、苯乙醇胺N-甲基转移酶、单胺氧化酶B等。结论:可视化针刀能缓解ONFH疼痛触发点比格犬模型的症状,其机制可能与调节神经递质代谢相关基因、生物学过程以及信号通路有关。
Objective:To observe the effect of visual needle knife therapy on myofascial trigger point of the beagle dog model with osteonecrosis of the femoral head(ONFH),and to explore the mechanism based on transcriptomics.Methods:A total of 24 beagle dogs were randomly divided into a control group,a model group,a visual needle knife therapy group and a traditional needle knife therapy group,with 6 dogs in each group.Except for the control group,the other beagle dogs were established ONFH myofascial trigger point model.The treatment was given once a week for 3 weeks in the visual needle knife therapy group and the traditional needle knife therapy group respectively.The serum levels of substance P(SP),5-hydro-xytryptamine(5-HT)and neuropeptide Y(NPY),calciyonon gene related peptide 1(CGRP1)and nitric oxide(NO)were detected by ELISA.HE staining was used to observe the pathological morphology around the hip joint.High-throughput sequencing technology(RNA-seq)was used to detect differentially expressed genes(DEGs),then Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis was performed.The ONFH neurotransmitter metabolic network was constructed using the STRING database and Cytoscape software,followed by identification of core targets through the intersection analysis of related genes and DEGs.Results:Hip CT showed that there was no abnormality in the hip joint of the beagle dogs in the control group.In the model group,the hip joint of the beagle dogs was in position,and a low-density area was observed in the femoral head.MRI of the hip joint showed that there was a small amount of effusion in the hip joint of the beagle dogs in the control group.In the model group,there was effusion in the hip joint,double line sign and mixed signal in the femoral head,accompanied by bone marrow edema.Before treatment,compared with the control group,the serum contents of SP,5-HT and NPY were increased,while that of NO and CGRP were decreased in the model group,traditional needle knife therapy group and visual needle knife therapy group significantly(P<0.05).Compared with that before treatment,after 21 days of treatment,the contents of SP,5-HT and NPY in the traditional needle knife therapy group and the visual needle knife therapy group were decreased,while the contents of NO and CGRP were increased significantly(P<0.05).After 21 days of treatment,compared with the traditional needle knife therapy group,the contents of SP,5-HT and NPY in the serum of beagle dogs in the visual needle knife therapy group were decreased,while the contents of NO and CGRP were increased significantly(P<0.05).HE staining showed that in the control group,the cytoplasm was evenly stained and the nuclei were arranged neatly.In the model group,the nuclei were arranged disorderly,the muscle fibers were loose and broken,the muscle stripes were blurred,and the local connective tissue proliferation was observed.In the traditional needle knife therapy group,the nuclei were arranged slightly disorderly,the local muscle striation was blurred,and the connective tissue was slightly hyperplasia.In the visual needle knife therapy group,the nuclei were arranged neatly and the muscle fibers were arranged closely.The results of transcriptome showed that 929 genes were down-regulated in the model group compared with the control group.Compared with the model group,3265 genes were up-regulated in the visual needle knife therapy group,and 685 genes in the two groups overlapped with each other.Compared with the control group,272 genes were up-regulated in the model group.Compared with the model group,748 genes were down-regulated in the visual needle knife therapy group,and 142 intersection genes were found between the two groups.The results of GO function analysis mainly involved positive regulation of antigen processing and presentation and phosphatidylinositol 3-kinase complex(class IB),sphingomyelin phosphodiesterase activity,etc.Related pathways of KEGG mainly included Osteoclast differentiation,Rheumatoid arthritis,Phagosome and so on.The core targets include acetylcholinesterase(ACHE),phenylethanolamine N-methyltransferase(PNMT),monoamine oxidase B(MAOB),etc.Conclusion:The visual needle knife therapy can relieve the symptoms of myofascial trigger point of the beagle dog model with ONFH,and its mechanism may be related to the regulation of neurotransmitter metabolism-related genes,biological processes and signaling pathways.
作者
孙诗艺
颜炎
何海军
吴子轩
王超
张程
张光熠
SUN Shiyi;YAN Yan;HE Haijun;WU Zixuan;WANG Chao;ZHANG Cheng;ZHANG Guangyi(Wangjing Hospital Affiliated to China Academy of Chinese Medical Sciences,Beijing China 100102;The Third Affiliated Hospital to Beijing University of Chinese Medicine,Beijing China 100029;Graduate School of Beijing University of Chinese Medicine,Beijing China 100029;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing China 100102)
出处
《中医学报》
CAS
2024年第9期1965-1973,共9页
Acta Chinese Medicine
基金
国家自然科学基金面上项目(81873322)。
关键词
可视化针刀
股骨头坏死
疼痛触发点
转录组学
神经递质
比格犬
visual needle knife therapy
osteonecrosis of the femoral head
myofascial trigger point
transcriptomics
neurotransmitter
beagle dog
