摘要
目的观察针刺“足三里”或“内关”对溃疡性结肠炎(Ulcerative colitis,UC)模型小鼠的干预效应,并利用转录组学分析病变组织结肠和非病变组织心脏在接受针刺干预后基因转录表达的异同之处,以期探讨两个腧穴是否存在功效差异。方法硫酸葡聚糖钠盐(Dextran sulfate sodium salt,DSS)诱导法建立UC小鼠模型,造模成功后,小鼠被随机分为模型组,足三里组及内关组,每组9只。足三里组及内关组分别于“足三里”、“内关”进行针刺干预,每次30 min,每天1次,连续干预5天,空白组和模型组只捆绑不针刺。观察各组小鼠的一般情况,计算疾病活动指数(Disease activity index,DAI),通过HE染色观察结肠组织形态变化,并利用转录组测序技术,分别对各组小鼠的结肠组织进行测序和生物学分析,从病变组织角度比较研究两个穴位是否存在腧穴功效差异。同时取各组心脏组织进行比较分析,从非病变组织角度探讨两个穴位的效应是否存在差异。结果与空白组相比,模型组DAI评分显著升高(P<0.001),HE染色结果显示伴有明显病理损伤;针刺足三里或内关后,UC小鼠的DAI评分均显著降低(P<0.05),病理损伤明显改善。转录组测序及生物信息学分析结果显示,针刺足三里或内关对病变部位结肠组织的影响涉及多个基因,相似变化的差异基因(co-DEGs)有825个,主要富集到白细胞及趋化因子等通路;只受足三里影响的差异基因(ST36only-DEGs)有168个,富集到胃酸分泌等通路;只受内关影响的差异基因(PC6 only-DEGs)有325个,富集到细胞因子-细胞因子受体相互作用等通路。非病变组织心脏中co-DEGs为95个,主要富集到防御反应的正向调节、对白细胞介素-1的反应等通路;ST36 only-DEGs为28个;PC6 only-DEGs为114个,富集到心血管疾病等通路。结论针刺“足三里”与“内关”均能改善UC小鼠的症状和结肠病理变化。但通过转录组学分析发现,两个穴位对病变部位结肠组织和非病变部位心脏组织的基因转录影响存在一定差异,提示分布部位不同的腧穴在宏观功效上有一定共性,但在微观层面上,存在一定的差异性。
Objective To observe the intervention effect of acupuncture at"Zusanli"(ST36)or"Neiguan"(PC6)on the ulcerative colitis(UC)model mice.And to analyze the similarities and differences of gene transcription expression between the diseased colon and the non-diseased heart after acupuncture intervention by using transcriptomics,to explore whether there are efficacy differences between the two acupoints.Methods The UC mouse model was established by inducing dextran sulfate sodium salt(DSS).After successful modeling,the mice were randomly divided into model group,Zusanli group,and Neiguan group,with 9 mice in each group.Zusanli group and Neiguan group intervened with acupuncture at"Zusanli"and"Neiguan",respectively,30 min each time,once a day,for 5 consecutive days.The control and model groups were only bound without acupuncture.Observe the general situation of mice in each group,calculate the disease activity index(DAI),observe the morphological changes of colon tissue by HE staining.Use transcriptome sequencing technology to sequence and biologically analyze the colon tissue of mice in each group,and compare and study whether the two acupoints have differences in acupoint efficacy from the perspective of diseased tissue.At the same time,take the heart tissues of each group for comparative analysis,and discuss whether the effects of the two acupoints are different from the perspective of non-diseased tissue.Results Compared with control group,the DAI score of the model group increased significantly(P<0.001),and HE staining results showed significant pathological damage;after acupuncture at Zusanli or Neiguan,the DAI scores of UC mice were reduced considerably(P<0.05),and the pathological damage was significantly improved.The results of transcriptome sequencing and bioinformatics analysis showed that the effects of acupuncture at Zusanli or Neiguan on the colon of diseased tissue involved multiple genes,and 825 differential genes with similar changes(co-DEGs)were mainly enriched in white blood cells and chemokine pathways;there were 168 differentially expressed genes(ST36 only-DEGs)affected only by Zusanli,which were enriched to gastric acid secretion and other pathways;there were 325 differentially expressed genes(PC6 only-DEGs)only affected by Neiguan,which were enriched in cytokine-cytokine receptor interaction and other pathways.There were 95 co-DEGs in the heart of non-diseased tissue,which were mainly enriched in positive regulation of defense response and response to interleukin-1;there were 28 ST36 only-DEGs;there were114 PC6 only-DEGs,which were enriched in cardiovascular diseases and other pathways.Conclusion Acupuncture of“Zusanli”and“Neiguan”could improve UC mice's symptoms and pathological colon changes.However,through transcriptomic analysis,it was found that the two acupoints had differences in gene transcription effects on the colon of diseased tissue and the heart of non-diseased tissue,suggesting that acupoints with different distribution positions had some similarities in macro efficacy.Still,there were some differences at a micro level.
作者
董龙聪
张瑞斌
沈缘
李虹莹
黄琴
余曙光
吴巧凤
Dong Longcong;Zhang Ruibin;Shen Yuan;Li Hongying;Huang Qin;Yu Shuguang;Wu Qiaofeng(Acupuncture and Tuina College,Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2024年第1期119-128,共10页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家科学技术部重点研发计划“中医药现代化研究”重点专项课题(2019YFC1709001):腧穴“特效”、“共效”的功能特点研究,负责人:余曙光
国家科学技术部重点研发计划“中医药现代化研究”重点专项课题(2022YFC3500703):刺激量效关系、腧穴同功效应及其配伍机制研究,负责人:吴巧凤
国家自然科学基金委员会面上基金(82174512):重塑KYN代谢途径介导的中枢胶质细胞稳态:艾灸改善溃疡性结肠炎负性情绪的关键机制,负责人:吴巧凤
作者简介
通讯作者:吴巧凤,研究员,博士研究生导师,主要研究方向:针灸调整神经-内分泌-免疫机制的研究。
