摘要
目的:基于线粒体自噬-核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症体探讨铁皮石斛抗动脉粥样硬化(AS)的分子机制。方法:选取6只C57BL/6J小鼠为对照组,将18只载脂蛋白E基因敲除(ApoE^(-/-))小鼠随机分为AS组、铁皮石斛组、铁皮石斛+线粒体分裂抑制剂-1(Mdivi-1)组,每组6只。检测各组小鼠血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白(LDL)水平;苏木精-伊红(HE)染色检测小鼠胸主动脉病理结构;油红O染色检测小鼠胸主动脉脂质浸润;蛋白免疫印迹法(Western Blot)检测线粒体自噬相关蛋白Beclin1、自噬微管相关蛋白轻链3Ⅱ(LC3Ⅱ)及NLRP3炎症体蛋白水平;实时荧光定量聚合酶链式反应(RT-qPCR)检测Beclin1、LC3Ⅱ及NLRP3炎症体mRNA水平;免疫组织化学染色检测小鼠胸主动脉NLRP3蛋白水平;检测各组小鼠血清炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-18(IL-18)、白细胞介素-1β(IL-1β)水平。结果:HE染色结果显示,与对照组相比,AS组小鼠胸主动脉内膜增厚,结构紊乱,可见斑块形成;与AS组相比,铁皮石斛组小鼠胸主动脉结构异常改善。油红O染色结果显示,与对照组相比,AS组小鼠胸主动脉中可见大量脂质浸润;与AS组相比,铁皮石斛组小鼠胸主动脉脂质浸润改善。与对照组相比,AS组小鼠胸主动脉Beclin1、LC3Ⅱ蛋白及mRNA表达增加(P<0.05);与AS组相比,铁皮石斛组小鼠胸主动脉Beclin1、LC3Ⅱ蛋白及mRNA表达减少(P<0.05)。与对照组相比,AS组小鼠血清TC、TG、LDL水平升高(P<0.05),NLRP3蛋白及mRNA表达增加(P<0.05),血清TNF-α、IL-6、IL-18、IL-1β水平升高(P<0.05);与AS组相比,铁皮石斛组小鼠血清TC、TG、LDL水平降低(P<0.05),NLRP3蛋白及mRNA表达减少(P<0.05),血清TNF-α、IL-6、IL-18、IL-1β水平降低(P<0.05);与铁皮石斛组相比,铁皮石斛+Mdivi-1组小鼠血清TC、TG、LDL水平升高(P<0.05),NLRP3蛋白及mRNA表达增加(P<0.05),血清TNF-α、IL-6、IL-18、IL-1β水平升高(P<0.05)。结论:铁皮石斛可通过上调线粒体自噬水平进而下调NLRP3介导的炎症反应,发挥抗AS的作用。
Objective:To explore the molecular mechanism of Dendrobium officinale in anti-atherosclerosis(AS)based on mitochondrial autophagy-nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammatory body.Methods:Six C57BL/6J mice were selected as the control group.Eighteen ApoE-knockout(ApoE^(-/-))mice were randomly divided into the AS group,Dendrobium officinale group,and Dendrobium officinale+mitochondrial fission inhibitor-1(Mdivi-1)group,with 6 mice in each group.The levels of total cholesterol(TC),triglycerides(TG),and low-density lipoprotein(LDL)in the serum of each group of mice were measured.Hematoxylin-eosin(HE)staining was used to detect the pathological structure of the thoracic aorta in mice.Oil Red O staining was used to detect lipid infiltration in the thoracic aorta of mice.Western Blot was used to detect the levels of mitochondrial autophagy-related protein Beclin1,autophagy microtubule-associated protein light chain 3Ⅱ(LC3Ⅱ),and NLRP3 inflammasome protein.Real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA levels of Beclin1,LC3Ⅱand NLRP3 inflammasome.Immunohistochemical staining was used to detect the level of NLRP3 protein in the thoracic aorta of mice.The levels of inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-18(IL-18),and interleukin-1β(IL-1β)in the serum of each group of mice were detected.Results:Compared with the AS group,the levels of TC,TG,and LDL in the serum of mice in the Dendrobium officinale group decreased(P<0.05).The HE staining results showed that compared with the control group,the intima of the thoracic aorta in the AS group was thickened and disordered,with plaque formation observed;compared with the AS group,the structure of the thoracic aorta in the Dendrobium officinale group improved significantly.The oil red O staining results showed that,compared with the control group,a large amount of lipid infiltration could be observed in the thoracic aorta of the AS group mice;and compared with the AS group,the lipid infiltration in the thoracic aorta of the Dendrobium officinale group mice was improved.Compared with the control group,the expression of Beclin1,LC3Ⅱprotein and mRNA in the thoracic aorta of the AS group decreased(P<0.05);compared with the AS group,the expression of Beclin1,LC3Ⅱprotein and mRNA in the thoracic aorta of the Dendrobium officinale group increased(P<0.05).Compared with the control group,the levels of TC,TG and LDL in the AS group decreased(P<0.05),the expression of NLRP3 protein and mRNA increased(P<0.05),and the levels of TNF-α,IL-6,IL-18 and IL-1βin the serum increased(P<0.05).Compared with the AS group,the levels of TC,TG and LDL in the Dendrobium officinale group increased(P<0.05),the expression of NLRP3 protein and mRNA decreased(P<0.05),and the levels of TNF-α,IL-6,IL-18 and IL-1βin the serum decreased(P<0.05).Compared with the Dendrobium officinale group,the serum levels of TC,TG and LDL in Dendrobium officinale+Mdivi-1 group increased(P<0.05),the expression of NLRP3 protein and mRNA increased(P<0.05),and the levels of TNF-α,IL-6,IL-18 and IL-1βincreased(P<0.05).Conclusion:Dendrobium officinale can improve atherosclerosis by up-regulating mitophagy and down-regulating NLRP3-mediated inflammatory response.
作者
史玉洁
刘晓丽
韩俊成
史玉琴
SHI Yujie;LIU Xiaoli;HAN Juncheng;SHI Yuqin(Zhangjiakou First Hospital(Affiliated People's Hospital of Zhangjiakou College,Zhangjiakou Cancer Hospital),Zhangjiakou 075000,Hebei,China;Second Hospital of Zhangjiakou City;Zhangjiakou Xuanhua District Market Supervision and Administration Bureau Food Testing,Zhangjiakou 075000,Hebei,China)
出处
《中西医结合心脑血管病杂志》
2025年第19期2942-2948,共7页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
河北省卫生健康科研基金项目(No.2021034)
张家口市重点研发计划项目(No.2322130D)。
关键词
动脉粥样硬化
铁皮石斛
线粒体自噬
核苷酸结合寡聚化结构域样受体蛋白3
血脂
炎性因子
实验研究
atherosclerosis
Dendrobium officinale
mitochondrial autophagy
nucleotide-binding oligomerization domain-like receptor protein 3
blood lipids
inflammatory factors
experimental study
