基于5-LOX/LTB4信号通路探讨黄芩苷/栀子苷对PM_(2.5)暴露致血管内皮功能障碍的干预作用及其机制  

Intervention of baicalin/geniposide on vascular endothelial dysfunction caused by PM_(2.5)exposure and its mechanism based on 5-LOX signaling pathway

作　　者：孙欠欠 张行行 赵麓 赵安东 史永恒[1,2,3] 王川 刘继平 王斌[1,2,3] SUN Qian-qian;ZHANG Hang-hang;ZHAO Lu;ZHAO An-dong;SHI Yong-heng;WANG Chuan;LIU Ji-ping;WANG Bin(School of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046;Shaanxi University Engineering Research Center of Traditional Chinese Medicine Brain Health Industry,Xianyang Shaanxi 712046;Key Laboratory of Pharmacodynamic Mechanism and Material Base of Traditional Chinese Medicine,Shaanxi Administration Bureau of Traditional Chinese Medicine,Xianyang Shaanxi 712046)

机构地区：[1]陕西中医药大学药学院,陕西咸阳712046 [2]中医药脑健康产业陕西省高校工程研究中心,陕西咸阳712046 [3]陕西省中医药管理局中药药效机制与物质基础重点研究室,陕西咸阳712046

出　　处：《中南药学》2024年第2期307-314,共8页Central South Pharmacy

基　　金：中医药“双链融合”中青年科研创新团队(No.2022-SLRH-YQ-006);国家自然科学基金项目(No.81473385);陕西省中医药科研项目(No.2021-ZZ-JC023);咸阳市重点研发科技项目(No.JBGS-001)。

摘　　要：目的基于5-LOX/LTB4信号通路探究黄芩苷/栀子苷(BC/GD)对PM_(2.5)诱导的血管内皮功能障碍的改善作用及其机制。方法利用离体肌张力描记技术测定BC/GD对不同状态下血管环张力的影响。将32只大鼠随机分为对照组、PM_(2.5)组、30 mg·kg^(-1)BC/GD组和60 mg·kg^(-1)BC/GD组,利用气管滴注法构建PM_(2.5)暴露模型,持续2个月,造模1个月后灌胃给予对应药物,持续1个月,末次给药后,HE染色观察肠系膜动脉血管内皮状态;化学发光法检测肠系膜动脉活性氧(ROS)水平;硝酸还原酶法检测一氧化氮(NO)水平;ELISA法检测血清炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、转化生长因子β1(TGF-β1)、白三烯B4(LTB4)水平;Western blot法检测5-脂氧酶(5-LOX)、内皮细胞一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)蛋白表达。结果BC/GD对基础状态的肠系膜动脉血管环张力无明显影响,但能明显舒张由5-HT预收缩的血管环;一氧化氮合酶抑制剂L-NAME、环氧合酶抑制剂INDO或L-NAME+INDO均能明显抑制BC/GD的血管舒张作用,但L-NAME的抑制作用更明显;50、100 mg·mL^(-1)PM_(2.5)能降低血管环对乙酰胆碱(ACh)的舒张血管效应,而BC/GD能明显改善PM_(2.5)诱导的舒张效应减弱。与对照组相比,PM_(2.5)组大鼠肠系膜动脉内皮完整性严重受损、内皮皱缩,大鼠血清中TNF-α、IL-1β、IL-6、LTB4水平显著升高、TGF-β1水平显著降低,肠系膜动脉组织中5-LOX、iNOS蛋白表达明显增加,eNOS蛋白表达降低,ROS水平升高,NO水平降低。与PM_(2.5)组相比,BC/GD能改善内皮损伤程度和完整性,可显著降低血清中TNF-α、IL-1β、IL-6、LTB4水平,升高TGF-β1水平;明显降低肠系膜动脉5-LOX、iNOS表达,升高eNOS蛋白表达;降低ROS水平,升高NO水平。结论BC/GD可通过抑制5-LOX/LTB4信号通路表达,从而降低ROS水平,抑制炎性损伤,上调eNOS表达,下调iNOS表达,升高血管中NO�Objective To determine the ameliorative effect of baicalin/geniposide(BC/GD)on PM_(2.5)-induced vascular endothelial dysfunction and its mechanism based on 5-LOX/LTB4 signaling pathway.Methods The effect of BC/GD on vascular ring tone in different states was determined with an ex vivo muscle tone tracing technique.Thirty-two rats were randomly divided into a control group,a PM_(2.5)group,a 30 mg·kg^(-1)BC/GD group and a 60 mg·kg^(-1)BC/GD group.The exposure model of PM_(2.5)was established by tracheal instillation for 2 months,and corresponding drugs were given by gavage for 1 month after the model establishment.After the last administration,HE staining was used to observe the endothelial status of mesenteric artery vasculature.Chemiluminescence detected the level of ROS in the mesenteric arteries;nitrate reductase detected the level of NO;serum levels of inflammatory factors TNF-α,IL-1β,IL-6,TGF-β1,and LTB4 were detected by ELISA;and Western blot detected the levels of 5-LOX,eNOS,and iNOS protein expression.Results BC/GD had no significant effect on the mesenteric artery vascular ring tone in the base state,but it significantly diastoled the vascular ring preconstricted by 5-HT.The nitric oxide synthase inhibitor L-NAME,the cyclooxygenase inhibitor INDO,or L-NAME+INDO all significantly inhibited the vasodilatory effect of BC/GD,and the inhibitory effect of L-NAME was more obvious;PM_(2.5)at 50 mg·mL-1 and 100 mg·mL-1 reduced the vasodilatory effect of vascular ring on acetylcholine,whereas BC/GD significantly ameliorated the PM_(2.5)-induced attenuation of the diastolic effect.The endothelial integrity of the mesenteric arteries was severely impaired and the endothelium was wrinkled in the PM_(2.5)group.Compared with those of the control group,the serum levels of TNF-α,IL-1β,IL-6,and LTB4 were higher and the level of TGF-β1 much lower in the PM_(2.5)group,and the protein expression of 5-LOX and iNOS was significantly increased and the expression of eNOS was decreased in the tissue of mesenteric arterie

关 键 词：PM_(2.5) 黄芩苷 栀子苷 内皮功能障碍 5-LOX/LTB4信号通路 

分 类 号：R285.5[医药卫生—中药学]