Stigmasterol is a plant sterol with anti-apoptotic,anti-oxidative and anti-inflammatory effect through multiple mechanisms.In this study,we further assessed whether it exerts protective effect on human brain microvess...Stigmasterol is a plant sterol with anti-apoptotic,anti-oxidative and anti-inflammatory effect through multiple mechanisms.In this study,we further assessed whether it exerts protective effect on human brain microvessel endothelial cells(HBMECs)against ischemia-reperfusion injury and explored the underlying mechanisms.HBMECs were used to establish an in vitro oxygen and glucose deprivation/reperfusion(OGD/R)model,while a middle cerebral artery occlusion(MCAO)model of rats were constructed.The interaction between stigmasterol and EPHA2 was detected by surface plasmon resonance(SPR)and cellular thermal shift assay(CETSA).The results showed that 10μmol·L−1 stigmasterol significantly protected cell viability,alleviated the loss of tight junction proteins and attenuated the blood-brain barrier(BBB)damage induced by OGD/R in the in vitro model.Subsequent molecular docking showed that stigmasterol might interact with EPHA2 at multiple sites,including T692,a critical gatekeep residue of this receptor.Exogenous ephrin-A1(an EPHA2 ligand)exacerbated OGD/R-induced EPHA2 phosphorylation at S897,facilitated ZO-1/claudin-5 loss,and promoted BBB leakage in vitro,which were significantly attenuated after stigmasterol treatment.The rat MCAO model confirmed these protective effects in vivo.In summary,these findings suggest that stigmasterol protects HBMECs against ischemia-reperfusion injury by maintaining cell viability,reducing the loss of tight junction proteins,and attenuating the BBB damage.These protective effects are at least meditated by its interaction with EPHA2 and inhibitory effect on EPHA2 phosphorylation.展开更多
苦参碱是来源于苦参等中药的一种天然喹啉类生物碱,具有广泛的药理作用。研究表明,苦参碱具有良好的抗肿瘤作用,作用机制主要与调控Wnt/β-连环蛋白、白细胞介素-6/转录激活因子3、核因子-κB、磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷...苦参碱是来源于苦参等中药的一种天然喹啉类生物碱,具有广泛的药理作用。研究表明,苦参碱具有良好的抗肿瘤作用,作用机制主要与调控Wnt/β-连环蛋白、白细胞介素-6/转录激活因子3、核因子-κB、磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白等信号通路密切相关。此外,苦参碱在抗炎、抗菌、抗病毒、神经保护、免疫调节等方面也显示良好的药理活性。肠吸收和药动学研究显示,苦参碱具有较高的溶解性和渗透性,药物吸收速度较快,通过将其制成纳米新制剂后,可进一步明显延长其半衰期并提高生物利用度。不良反应方面,苦参碱的相关研究主要集中在肝、肾损伤,在未来应用中控制用药剂量是降低其不良反应的有效方式,从而保证其用药的安全性与有效性。通过检索中国知网、维普、Web of Science等国内外数据库,对苦参碱的药理作用、肠吸收、药动学及不良反应进行综述,为苦参碱的进一步研究开发和临床应用提供借鉴。展开更多
基金supported by the Key Research Project of the Science&Technology Department of Sichuan Province,China(Nos.2021YFS0131 and 2020YFS0414).
文摘Stigmasterol is a plant sterol with anti-apoptotic,anti-oxidative and anti-inflammatory effect through multiple mechanisms.In this study,we further assessed whether it exerts protective effect on human brain microvessel endothelial cells(HBMECs)against ischemia-reperfusion injury and explored the underlying mechanisms.HBMECs were used to establish an in vitro oxygen and glucose deprivation/reperfusion(OGD/R)model,while a middle cerebral artery occlusion(MCAO)model of rats were constructed.The interaction between stigmasterol and EPHA2 was detected by surface plasmon resonance(SPR)and cellular thermal shift assay(CETSA).The results showed that 10μmol·L−1 stigmasterol significantly protected cell viability,alleviated the loss of tight junction proteins and attenuated the blood-brain barrier(BBB)damage induced by OGD/R in the in vitro model.Subsequent molecular docking showed that stigmasterol might interact with EPHA2 at multiple sites,including T692,a critical gatekeep residue of this receptor.Exogenous ephrin-A1(an EPHA2 ligand)exacerbated OGD/R-induced EPHA2 phosphorylation at S897,facilitated ZO-1/claudin-5 loss,and promoted BBB leakage in vitro,which were significantly attenuated after stigmasterol treatment.The rat MCAO model confirmed these protective effects in vivo.In summary,these findings suggest that stigmasterol protects HBMECs against ischemia-reperfusion injury by maintaining cell viability,reducing the loss of tight junction proteins,and attenuating the BBB damage.These protective effects are at least meditated by its interaction with EPHA2 and inhibitory effect on EPHA2 phosphorylation.
文摘苦参碱是来源于苦参等中药的一种天然喹啉类生物碱,具有广泛的药理作用。研究表明,苦参碱具有良好的抗肿瘤作用,作用机制主要与调控Wnt/β-连环蛋白、白细胞介素-6/转录激活因子3、核因子-κB、磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白等信号通路密切相关。此外,苦参碱在抗炎、抗菌、抗病毒、神经保护、免疫调节等方面也显示良好的药理活性。肠吸收和药动学研究显示,苦参碱具有较高的溶解性和渗透性,药物吸收速度较快,通过将其制成纳米新制剂后,可进一步明显延长其半衰期并提高生物利用度。不良反应方面,苦参碱的相关研究主要集中在肝、肾损伤,在未来应用中控制用药剂量是降低其不良反应的有效方式,从而保证其用药的安全性与有效性。通过检索中国知网、维普、Web of Science等国内外数据库,对苦参碱的药理作用、肠吸收、药动学及不良反应进行综述,为苦参碱的进一步研究开发和临床应用提供借鉴。
