[关键词]
[摘要]
【目的】探讨“温阳通脉”灸法防治动脉粥样硬化 (AS) 的作用机制。 【方法】将10只饲喂普通饲料的C57BL/6J小鼠设为 空白组;将30只ApoE-/- 小鼠给予高脂饲料喂养建立动脉粥样硬化模型,随机分为模型组、辛伐他汀组和艾灸组,每组10只。 于造模第1天开始干预,艾灸组小鼠给予膻中、神阙、内关、血海穴艾灸,辛伐他汀组给予辛伐他汀蒸馏水混悬液灌胃,共 干预12周。给药结束后,采用苏木素-伊红 (HE) 染色法观察小鼠胸主动脉病理形态结构,透射电镜观察小鼠胸主动脉内皮 细胞超微结构,酶联免疫吸附法 (ELISA) 检测小鼠血清肿瘤坏死因子α (TNF-α) 、细胞间黏附分子1 (ICAM-1) 、血管细胞黏 附分子1 (VCAM-1) 水平,实时定量聚合酶链反应 (qRT-PCR) 法检测胸主动脉沉默信息调节因子1 (SIRT1) 和叉头框转录因子 O3a (FOXO3a) 的mRNA表达水平,Western Blot法检测胸主动脉SIRT1和FOXO3a蛋白表达水平。 【结果】与空白组比较,模 型组小鼠胸主动脉及血管内皮细胞病理变化明显,血清炎症因子TNF-α、ICAM-1和VCAM-1水平升高 (P<0.05或P< 0.01) ,胸主动脉SIRT1 mRNA和蛋白表达量降低 (P<0.01) ,FOXO3a mRNA和蛋白表达量差异无统计学意义 (P>0.05) ;与 模型组比较,辛伐他汀组和艾灸组小鼠的胸主动脉及血管内皮细胞结构得到明显改善,血清TNF-α、ICAM-1、VCAM-1水 平均降低 (P<0.05) ,胸主动脉SIRT1 mRNA和蛋白表达量增加 (P<0.05或P<0.01) ,FOXO3a mRNA和蛋白表达量差异均无 统计学意义 (P>0.05) 。辛伐他汀组和艾灸组上述指标组间比较,差异均无统计学意义 (P>0.05) 。 【结论】 “温阳通脉”灸法 可通过调控SIRT1/FOXO3a信号通路减轻炎症反应防治小鼠AS。
[Key word]
[Abstract]
Objective To explore the mechanism of ‘Wenyang Tongmai’(warming yang to unblock meridians) moxibustion in preventing and treating atherosclerosis. Methods Ten C57BL/6J mice fed with normal diet were set as blank group. Thirty ApoE-/- mice were fed with high-fat diet to establish atherosclerosis model,and were randomly divided into model group,Simvastatin group and moxibustion group,with 10 mice in each group. The intervention began on the first day of modeling. The mice in the moxibustion group were given moxibustion at Danzhong (RN17),Shenque(RN8),Neiguan(PC6),Xuehai(SP10) points,and the Simvastatin group was given Simvastatin distilled water suspension by gavage for 12 weeks. After administration, the pathological structure of thoracic aorta in mice was observed by hematoxylin-eosin (HE) staining method. The ultrastructure of thoracic aortic endothelial cells in mice was observed by transmission electron microscopy. The levels of serum tumor necrosis factor α(TNF- α), intercellular adhesion molecule 1(ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) in mice were detected by enzyme-linked immunosorbent assay (ELISA) . The mRNA expression levels of silent information regulator 1 (SIRT1) and forkhead box O3a (FOXO3a) in thoracic aorta were detected by real-time quantitative polymerase chain reaction (qRT-PCR) . The protein expression levels of SIRT1 and FOXO3a in thoracic aorta were detected by Western Blot. Results Compared with the blank group,the pathological changes of thoracic aorta and vascular endothelial cells in the model group were obvious,the levels of serum inflammatory factor TNF- α,ICAM-1 and VCAM-1 were increased (P<0.05 or P<0.01),the mRNA and protein expressions of SIRT1 in thoracic aorta were decreased(P<0.01), and the mRNA and protein expressions of FOXO3a had no significant difference(P>0.05) . Compared with the model group,the thoracic aorta and vascular endothelial cell structure of the mice in the Simvastatin group and the moxibustion group were obviously improved,the levels of serum TNF-α,ICAM-1 and VCAM-1 were decreased (P<0.05) ,the mRNA and protein expressions of SIRT1 in the thoracic aorta were increased (P<0.05 or P<0.01) ,and the mRNA and protein expressions of FOXO3a had no significant difference(P>0.05) . There was no significant difference in the above indexes between the Simvastatin group and the moxibustion group (P>0.05) . Conclusion ‘Wenyang Tongmai’ moxibustion can prevent and treat atherosclerotic in rats via regulating and controlling SIRT1/FOXO3a signaling pathway to reduce inflammatory response.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金资助项目 (编号:82160941,82360978);贵州省科技计划项目 (编号:黔科合基础-ZK[2022]一般 499);贵州省 卫生健康委科学技术基金资助项目 (编号:gzwkj2023-019)