香草酸对大鼠离体肺动脉血管的舒张作用及舒张肺动脉高压大鼠体内肺动脉血管的机制  

Vasodilatory effects of vanilla acid on isolated pulmonary artery in rats and the mechanism of relaxation of pulmonary artery in PAH rats

作　　者：王良奇 杨可 谢新 孙蒙蒙 李占强[1,2] 芦殿香 WANG Liangqi;YANG Ke;XIE Xin;SUN Mengmeng;LI Zhanqiang;LU Dianxiang(Research Center for High Altitude Medicine,Key Laboratory of Ministry of Education for High Altitude Medicine,Laboratory for High Altitude Medicine in Qinghai Province,Key Laboratory of Application and Foundation for High Altitude Medicine Research in Qinghai Province(Qinghai-Utah Joint Research Key Lab for High Altitude Medicine),Qinghai University,Xining 810001,China;College of Eco-environmental Engineering,Qinghai University,Xining 810016,China;Clinical Medical ColleAfiliated Hospital of Chengdu University,Chengdu 610081,China)

机构地区：[1]青海大学高原医学研究中心,高原医学教育部重点实验室,青海省高原医学实验室,青海省高原医学应用基础重点实验室(青海-犹他高原医学联合重点实验室),西宁810001 [2]青海大学生态环境工程学院,西宁810016 [3]成都大学附属医院,成都610081

出　　处：《中国高原医学与生物学杂志》2024年第4期246-254,共9页Journal of Chinese High Altitude Medicine & Biology

基　　金：国家自然科学基金资助项目(82360831);青海省高端创新人才计划项目。

摘　　要：目的探讨香草酸对大鼠离体肺动脉血管的舒张作用及舒张肺动脉高压(PAH)大鼠体内肺动脉血管的机制。方法利用离体血管张力测定系统评价香草酸对大鼠离体肺小动脉血管环的舒张作用。并使用格列苯脲(GLB)、N-硝基-L-精氨酸甲酯盐酸盐(L-NAME)、4-氨基吡啶(4-AP)、吲哚美辛(IMC)、氯化钡(BaCl2)和四乙基铵(TEA)等工具药物明确香草酸舒张大鼠离体肺动脉血管的作用机制。并进一步在由野百合碱(MCT)诱导的PAH大鼠体内,利用Western Blot技术检测肺组织中与NO合成相关的蛋白质(eNOS、AMPK)的表达情况。应用HPLC技术测定大鼠肺组织中的AMP、ADP、ATP含量。采用分子对接法验证香草酸与eNOS、AMPK之间的结合情况。结果香草酸能够有效舒张由NE预收缩的血管环,EC50为137.9μmol/L。L-NAME能有效阻断香草酸的舒血管效应。体内研究显示,香草酸能有效提高MCT诱导的PAH大鼠肺组织中eNOS、AMPK蛋白的表达水平及磷酸化水平,且能够提高(ADP+AMP)/ATP。分子对接结果显示,香草酸同eNOS、AMPK的结合能分别为-6.1 kcal/mol、-6.0 kcal/mol。结论香草酸具有舒张大鼠离体肺动脉血管的作用;香草酸能够通过调节(ADP+AMP)/ATP,激活AMPK/eNOS轴,舒张PAH大鼠体内肺动脉血管,干预PAH的病理发展。Objective To investigate the vasodilatory effects of vanillic acid(VA)on isolated pulmonary artery in rats and the mechanism of relaxation of pulmonary artery in PAH rats.Methods The vasodilatory effects of VA on the vascular ring of isolated pulmonary arterioles in rats were assessed using an in vitro vascular tension measurement system.The mechanism of action of VA in relaxing isolated pulmonary arteries in rats is clarified by using drugs such as glibenclamide(GLB),N-nitro-L-arginine methyl ester hydrochloride(L-NAME),4-aminopyridine(4-AP),indomethacin(IMC),barium chloride(BaCl,)and tetraethylammonium(TEA).Western blotting was utilized to examine the expression of nitric oxide(NO)synthesis-related proteins(eNOS and AMPK)in lung tissue from rats with monocrotaline-induced pulmonary hypertension(PAH).The levels of AMP,ADP and ATP in lung tissue of rats were determined via HPLC analysis.Furthermore,molecular docking was employed to validate the binding interaction between VA and eNOS as well as AMPK.Results VA could significantly relax the vascular rings pre-contracted by NE with an ECso value of 137.9μM.The vasodilatory effects of VA were effectively blocked by L-NAME.In vivo studies demonstrated that VA could effectively upregulate the expression and phosphorylation levels of eNOS and AMPK proteins in lung tissue of McT-induced PAH rats,and increase the levels of(ADP+AMP)/ATP.The molecular docking results showed that the binding energies of VA with eNOS and AMPK were-6.1 kcal/mol and-6.0 kcal/mol respectively Conclusion Vanillic acid can relax isolated pulmonary artery blood vessels in rats.In addition,it can regulate the levels of(ADP+AMP)/ATP and activate the AMPK/eNOS axis to relax the pulmonary arteries in PAH rats,thereby intervening in the pathological development of PAH.

关 键 词：香草酸 血管 ENOS ATP 分子对接 

分 类 号：R338[医药卫生—人体生理学]