摘要
探究峨参总提物对肺动脉高压大鼠的作用效果及作用机制。将60只雄性SD大鼠分为正常(NC)组、模型(M)组、阳性药西地那非(Y)组、峨参低剂量(ES-L)组、峨参中剂量(ES-M)组、峨参高剂量(ES-H)组。大鼠在第1天腹腔注射野百合碱(60 mg·kg^(-1)),28 d形成肺动脉高压大鼠模型,在第15~28天灌胃给药治疗。造模及给药结束后,用小动物超声成像检测大鼠右心功能;血气分析仪检测大鼠动脉血气;苏木素-伊红(HE)染色及马松染色观察大鼠心肺病理损伤;流式细胞术检测大鼠肺组织及心肌组织凋亡及活性氧(ROS)水平;Western blot法检测肺组织转化生长因子β1(TGF-β1)、磷酸化母亲DPP同源物3(p-Smad3)、母亲DPP同源物3(Smad3)、纤维溶酶原激活物(t-PA)、纤溶酶原激活物抑制物1(PAI-1)表达水平;血常规检测仪检测大鼠血液炎症免疫细胞水平;酶联免疫吸附测定(ELISA)法检测血浆中P-选择素、血栓素A2(TXA2)表达水平。结果显示,与NC组相比,M组大鼠右心肥厚指数、右心室游离壁厚度以及右心内径、二氧化碳分压(PaCO_(2))以及心肺组织细胞凋亡、ROS水平显著升高,肺动脉血流加速时间(PAT)/射血时间(PET)的比值、右心输出量、右心室收缩面积变化率、三尖瓣环收缩期位移、氧分压(PaO_(2))以及血氧饱和度(SaO_(2))显著降低;给药峨参总提物后,大鼠右心功能、血气水平显著升高,心肺组织凋亡及ROS水平显著降低。进一步检测发现,峨参总提物能够显著降低肺组织白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、PAI-1蛋白表达水平,升高t-PA蛋白表达水平,减少血液中白细胞、淋巴细胞、粒细胞、单核细胞等炎症细胞水平以及血浆中P-选择素、TXA2水平。代谢组学结果显示,峨参总提物显著影响的代谢通路主要包括精氨酸生物合成、酪氨酸代谢、牛磺酸和亚牛磺酸代谢等。因此,峨参总提物可能通过抑制TGF-β1/Smad3信号通路减轻大鼠免疫炎症和血栓形成来发挥抗肺动脉高压的作用。
This study aimed to explore the effects and mechanisms of total extracts from Anthriscus sylvestris on pulmonary hypertension in rats.Sixty male SD rats were divided into normal(NC)group,model(M)group,positive drug sildenafil(Y)group,low-dose A.sylvestris(ES-L)group,medium-dose A.sylvestris(ES-M)group,and high-dose A.sylvestris(ES-H)group.On day 1,rats were intraperitoneally injected with monocrotaline(60 mg·kg^(-1))to induce pulmonary hypertension,and the rat model was established on day 28.From days 15 to 28,intragastric administration of the respective treatments was performed.After modeling and treatment,small animal echocardiography was used to detect the right heart function of the rats.Arterial blood gas was measured using a blood gas analyzer.Hematoxylin and eosin(HE)staining and Masson staining were performed to observe cardiopulmonary pathological damage.Flow cytometry was used to detect apoptosis in the lung and myocardial tissues and reactive oxygen species(ROS)levels.Western blot was applied to detect the expression levels of transforming growth factor-β1(TGF-β1),phosphorylated mothers against decapentaplegic homolog 3(p-Smad3),Smad3,tissue plasminogen activator(t-PA),and plasminogen activator inhibitor-1(PAI-1)in lung tissue.A blood routine analyzer was used to measure inflammatory immune cell levels in the blood.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression levels of P-selectin and thromboxane A2(TXA2)in plasma.The results showed that,compared with the NC group,right heart hypertrophy index,right ventricular free wall thickness,right heart internal diameter,partial carbon dioxide pressure(PaCO_(2)),apoptosis in cardiopulmonary tissue,and ROS levels were significantly increased in the M group.In contrast,the ratio of pulmonary blood flow acceleration time(PAT)/ejection time(PET),right cardiac output,change rate of right ventricular systolic area,systolic displacement of the tricuspid ring,oxygen partial pressure(PaO_(2)),and blood oxygen saturation(SaO_(2))were significantly decreased in the M group.After administration of the total extract of A.sylvestris,right heart function and blood gas levels were significantly improved,while apoptosis in cardiopulmonary tissue and ROS levels significantly decreased.Further testing revealed that the total extract of A.sylvestris significantly decreased the levels of interleukin-1β(IL^(-1)β),interleukin-6(IL-6),and PAI-1 proteins in lung tissue,while increasing the expression of t-PA.Additionally,the extract reduced the levels of inflammatory cells such as leukocytes,lymphocytes,granulocytes,and monocytes in the blood,as well as the levels of P-selectin and TXA2 in plasma.Metabolomics results showed that the total extract of A.sylvestris significantly affected metabolic pathways,including arginine biosynthesis,tyrosine metabolism,and taurine and hypotaurine metabolism.In conclusion,the total extract of A.sylvestris may exert an anti-pulmonary hypertension effect by inhibiting the TGF-β1/Smad3 signaling pathway,thereby alleviating immune-inflammatory responses and thrombosis.
作者
郑亚娟
袁培培
张振凯
刘晏灵
李赛菲
阮元
陈祎
傅阳
冯卫生
郑晓珂
ZHENG Ya-juan;YUAN Pei-pei;ZHANG Zhen-kai;LIU Yan-ling;LI Sai-fei;RUAN Yuan;CHEN Yi;FU Yang;FENG Wei-sheng;ZHENG Xiao-ke(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Engineering Technology Research Center for Traditional Chinese Medicine Development,Zhengzhou 450046,China;Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment&Chinese Medicine Development of Henan Province,Zhengzhou 450046,China)
出处
《中国中药杂志》
北大核心
2025年第9期2472-2483,共12页
China Journal of Chinese Materia Medica
基金
中央引导地方科技发展专项(豫财科[2016]149号)
国家重点研发计划中医药现代化研究项目(2019YFC1708802)
河南省高层次人才特殊支持计划“中原千人计划”中原领军人才项目(ZYQR201810080)
河南中医药大学博士科研基金项目(RSBSJJ2018-04)。
关键词
峨参
肺动脉高压
炎症
血栓形成
Anthriscus sylvestris
pulmonary arterial hypertension
inflammation
thrombosis
