香青兰总黄酮调控TMAO介导的JAK/STAT轴抗大鼠动脉粥样硬化及RAW264.7细胞炎症的研究  

Effect of total flavonoids of Dracocephalum moldavica on TMAO-mediated JAK/STAT axis against atherosclerosis in rats and inflammation in RAW264.7 cells

作　　者：曹文疆[1] 杜春妍 黄川生[1] 赵云丽 马晓莉 袁勇[1] 王新春[1] CAO Wen-jiang;DU Chun-yan;HUANG Chuan-sheng;ZHAO Yun-li;MA Xiao-li;YUAN Yong;WANG Xin-chun(The First Affiliated Hospital of Shihezi University,Shihezi Xinjiang 832008,China;Changji hui Autonomous Prefecture People's Hospital,Changji Xinjiang 831100,China)

机构地区：[1]石河子大学第一附属医院,新疆石河子832008 [2]昌吉回族自治州人民医院,新疆昌吉831100

出　　处：《中国药理学通报》2024年第9期1766-1772,共7页Chinese Pharmacological Bulletin

基　　金：兵团科技计划项目重点领域科技攻关计划(No 2022AB020);国家自然科学基金资助项目(No 81960766)。

摘　　要：目的探讨香青兰总黄酮(total flavonoids of Dracocephalum Moldavica L.,TFDM)抗大鼠动脉粥样硬化(atherosclerosis,AS)及三甲胺氮氧化物(trimethylamine N-oxide,TMAO)加剧的小鼠巨噬细胞RAW264.7炎症的保护作用及其可能的作用机制。方法采用高脂饲料喂养联合腹腔注射维生素D3的方法建立SD大鼠AS模型,分为对照组、模型组、辛伐他汀组(15 mg·kg^(-1))、TFDM组(60、30、15 mg·kg^(-1)),建模同时进行给药处理,持续8周。生化检测方法检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)水平。HE染色检测主动脉组织病理变化,ELISA试剂盒检测血清中TMAO、IL-1β、IL-6及肝脏组织中TNF-α的表达,Western blot法检测主动脉中JAK、STAT、TNF-α蛋白的表达。此外,体外培养RAW264.7巨噬细胞,采用LPS+TMAO建立巨噬细胞炎症模型,TFDM(100、50、25 mg·L^(-1))进行干预。CCK-8测定细胞活力及增殖,RT-qPCR法检测细胞中TNF-α、IL-6、JAK、STAT mRNA的表达。结果TFDM可明显下调大鼠血清TC、TG、LDL-C水平及血清TMAO、IL-1β、IL-6和肝脏TNF-α的水平,减少主动脉的斑块沉积,下调主动脉中TNF-α、JAK、STAT的蛋白表达。此外,TFDM干预可明显下调TNF-α、IL-6、JAK、STAT mRNA的表达及JAK、STAT蛋白的表达。结论TFDM可降低血清中TMAO的含量,从而抑制JAK/STAT炎症信号通路,减缓炎症的发生,发挥抗AS作用。Aim To investigate the protective effect of total flavonoids of Dracocephalum moldavica(TFDM)on atherosclerosis in rats and the inflammation of mouse macrophage RAW264.7 aggravated by trimethylamine N-oxide(TMAO)and its possible mechanism.Methods The AS model of SD rats was established by high-fat diet feeding combined with intraperitoneal injection of vitamin D3.The rats were divided into control group,model group,simvastatin group(15 mg·kg^(-1))and TFDM group(60,30,15 mg·kg^(-1)).Biochemical method was used to detect the levels of serum total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C).HE staining was used to detect the pathological changes of aortic tissue.ELISA kit was used to detect the expression of TMAO,IL-1β,IL-6 in serum and TNF-αin liver tissue.Western blot was used to detect the expression of JAK,STAT and TNF-αprotein in aorta.In addition,RAW264.7 macrophages were cultured in vitro,and LPS+TMAO was used to establish a macrophage inflammation model,which was intervened by TFDM(100,50,25 mg·L^(-1)).CCK-8 was used to determine cell viability and proliferation,and RT-qPCR was used to detect the expression of TNF-α,IL-6,JAK and STAT mRNA in cells.Results TFDM could significantly down-regulate the levels of serum TC,TG,LDL-C,serum TMAO,IL-1β,IL-6 and liver TNF-α,reduce aortic plaque deposition,and down-regulate the protein expression of TNF-α,JAK and STAT in aorta.In addition,TFDM intervention can significantly down-regulate the expression of TNF-α,IL-6,JAK,STAT mRNA and the expression of JAK,STAT protein.Conclusion TFDM can reduce the content of TMAO in serum,inhibit JAK/STAT inflammatory signaling pathway and slow down the occurrence of inflammation,playing an anti-AS role.

关 键 词：香青兰总黄酮 动脉粥样硬化 肠道菌群 氧化三甲胺 炎症反应 JAK/STAT信号通路 

分 类 号：R-332[医药卫生] R284.1R322.45R329.24R364.5R543.5