Effect of herb cake-insulated moxibustion on serum exosomal miR-223 expression in rabbits with atherosclerosis  

隔药饼灸对动脉粥样硬化兔血清外泌体miR-223表达的影响

作　　者：LI Qian PENG Han CHEN Xinyu YI Hongfen GE Junyun OUYANG Lizhi LIU Honghua LIU Mailan 李芊;彭涵;陈昕羽;易洪芬;葛君芸;欧阳里知;刘红华;刘迈兰(Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]Hunan University of Chinese Medicine,Changsha 410208,China

出　　处：《Journal of Acupuncture and Tuina Science》2025年第1期12-21,共10页针灸推拿医学(英文版)

基　　金：国家自然科学基金项目,No.81704182,No.82074559;长沙市杰出创新青年培养计划项目,No.kq1905036;湖南省青年科技创新人才项目,No.2022RC1222;2022年湖南中医药大学研究生创新课题,No.2022CX111。

摘　　要：Objective:To investigate whether herb cake-insulated moxibustion affects the expression of cholesterol metabolismrelated protein 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGR)and inhibits the development of atherosclerosis by regulating the exosomal miR-223 expression.Methods:Thirty-six male New Zealand rabbits were randomly assigned to a normal group,a model group,and an herb cake-insulated moxibustion group,with 12 rabbits in each group.The model and the herb cake-insulated moxibustion groups were fed a high-fat diet for 8 weeks to induce an atherosclerosis model.Following successful modeling,the herb cake-insulated moxibustion group was subjected to bundling and herb cake-insulated moxibustion intervention,while the other two groups were subjected only to bundling without moxibustion.After 8 weeks of intervention,hematoxylin-eosin(HE)staining was used to observe aortic morphology;the serum levels of total cholesterol(TC),triglyceride(TG),and low-density lipoprotein cholesterol(LDL-C)were detected using an automatic biochemical analyzer in each group.Exosome morphology was observed using the transmission electron microscope;Western blotting(WB)was used to detect the protein levels of serum exosomal CD63 and CD9 markers,as well as liver HMGR;additionally,real-time fluorescence quantitative polymerase chain reaction was used to quantify serum exosomal miR-223.Results:HE staining showed thickened aortic intima,lipid infiltration,foam cell aggregation,and structural damage to the arterial wall in the model group.Meanwhile,after modeling,the serum levels of LDL-C,TC,and TG increased significantly in the model and herb cake-insulated moxibustion groups compared to the normal group(P<0.05),suggesting successful atherosclerosis rabbit model preparation.The serum exosomes of rabbits in the model group exhibited a saucer-like or semi-concave spherical shape with diameters of 120-150 nm.WB detection results showed positive expression of the exosomal markers CD63 and CD9.After 8 weeks of intervention,the miR-223 lev目的:探讨隔药饼灸是否通过调控外泌体miR-223表达影响胆固醇代谢相关蛋白3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMGR)的表达,抑制动脉粥样硬化的发展。方法:将36只雄性新西兰兔随机均分为正常组、模型组和隔药饼灸组,每组12只。模型组和隔药饼灸组采用高脂饲料饲养8周制备动脉粥样硬化模型。模型成功后,隔药饼灸组捆绑后进行隔药饼灸干预;其他2组仅做相同的捆绑。干预8周后,采用苏木素-伊红(HE)染色观察主动脉形态;全自动生化仪检测各组血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)含量水平;透射电镜观察外泌体形态;免疫印迹(WB)法检测血清外泌体中CD63、CD9标志蛋白以及肝组织HMGR蛋白的表达量;实时荧光定量聚合酶链反应检测血清外泌体中miR-223的含量。结果:HE染色见模型组主动脉内膜增厚,脂质浸润、泡沫细胞聚集,动脉壁结构完整度受损。造模后,与正常组比较,模型组和隔药饼灸组动物血清中LDL-C、TC和TG含量显著升高(P<0.05),结合HE染色切片分析提示动脉粥样硬化兔模型制备成功。电镜观察模型组兔血清外泌体为茶托样状或半凹陷球状,直径为120-150 nm;WB检测结果显示蛋白标志物CD63、CD9在外泌体中表达呈阳性。干预8周后,与正常组相比,模型组miR-223表达显著下降(P<0.01);与模型组相比,隔药饼灸组血清中TC、TG及LDL-C含量显著降低(P<0.05),miR-223表达明显上升(P<0.01),肝组织HMGR蛋白相对含量下降(P<0.05)。结论:隔药饼灸可能是通过上调外泌体miR-223表达,下调HMGR蛋白表达来抑制胆固醇合成代谢,从而缓解动脉粥样硬化病程。

关 键 词：Moxibustion Therapy Herb Cake-insulated Moxibustion ATHEROSCLEROSIS 3-hydroxy-3-methylglutaryl Coenzyme A Reductase Embolism Cholesterol Exosomes RABBITS 

分 类 号：R2-03[医药卫生—中医学]