银杏蜜环口服溶液对缺氧缺糖再灌注诱导脑微血管内皮细胞和SH-SY5Y细胞炎性损伤的保护作用与机制
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摘要
目的:探讨银杏蜜环口服溶液对缺氧缺糖再灌注诱导脑微血管内皮细胞和神经胶质瘤细胞SH-HY5Y损伤的保护机制,以及其对e NOs介导的Beclin-1依赖的自噬通路的影响。方法:本研究采用缺氧缺糖再灌注诱导大鼠脑微血管内皮细胞和神经胶质瘤细胞SH-HY5Y构建糖氧剥离+复糖复氧模型,同时给予不同浓度的银杏蜜环口服溶液(5.15 mg/m L、2.575 mg/m L和1.545 mg/m L)及有效组分进行干预,通过观察细胞培养上清炎性因子IL-1β、TNF-α和IL-6的浓度和自噬通路的活化,并采用PI/Annexin V双染色分析细胞凋亡率,探讨银杏蜜环有效成份的脑保护机制。结果:银杏蜜环口服溶液5.15 mg/m L、2.575 mg/m L和1.545 mg/m L3个剂量组均可抑制脑微血管内皮细胞上清液TNF-α、IL-1β和IL-6含量,银杏蜜环口服溶液2.575 mg/m L和1.545 mg/m L可抑制Caspase3、LC3II和Beclin-1表达。同时,银杏蜜环口服溶液2.575 mg/m L明显促进e NOs的磷酸化。其中对L-1β和IL-6的作用明显优于银杏叶提取物和天麻蜜环菌。银杏蜜环口服溶液2.575 mg/m L和1.545 mg/m L还可降低SH-SY5Y细胞的凋亡率(Annexin V+/PI-细胞),差异有统计学意义。结论:银杏蜜环口服溶液可通过促进一氧化氮合酶e NOs的磷酸化来抑制缺糖缺氧再灌引发的细胞凋亡和自噬。
Objective: To study the protective effects and mechanisms of Yinxing Mihuan Oral Solution for cerebral microvascular endothelial cells( CMEC) and SH-SY5 Y cells on inflammatory induced by oxygen/glucose deprivation( OGD) and reperfusion,as well as the effects of Yinxing Mihuan oral solution on autophagy pathways mediated by endothelial nitric oxide synthase( e NOS)depended by Beclin-1. Methods: Oxygen/glucose deprivation and oxygen/glucose reintroduction model was established in murine cerebral microvascular endothelial cells and SH-SY5 Y cells by( OGD) and reperfusion. Meanwhile,different concentrations of Yinxing Mihuan Oral Solution( 5. 15 mg/m L,2. 575 mg/m L and 1. 545 mg/m L) were used,and efficient compositions were intervened. Through the observation of the concentrations of inflammatory factors IL-1 β,TNF-α and IL-6 in cell cultivated supernate and the excitation of autophagy pathways. Brain protective mechanism of Yinxing Mihuan active principles by apoptosis rate adopted the PI/Annexin V double staining analysis was discussed. Results: Yinxing Mihuan Oral Solution( 5. 15 mg/m L,2. 575 mg/m L and 1. 545 mg/m L) could restrain the contents of TNF-α,IL-1 β and IL-6 in cell culture supernatant of CMEC. Yinxing Mihuan Oral Solution( 2. 575 mg/m L) significantly promoted the phosphorylation of e NOs and Yinxing Mihuan Oral Solution( 2. 575 mg/m L and 1. 545 mg/m L) restrained the Caspase-3,LC3 II and Beclin-1 expression. The effects of L-1 β and IL-6 were significantly better than that of ginkgo biloba extract and mellea armillaria. Yinxing Mihuan Oral Solution( 2. 575 mg/m L and 1. 545 mg/m L) also decreased SH-SY5 Y cells apoptosis rate( Annexin V +/PI-cells),with significant difference. Conclusion: Yinxing Mihuan oral solution could promote the e NOs phosphorylation,which may be the interventional targets in restraining the cells' apoptosis and autophagy induced by OGD and reperfusion.
Objective: To study the protective effects and mechanisms of Yinxing Mihuan Oral Solution for cerebral microvascular endothelial cells( CMEC) and SH-SY5 Y cells on inflammatory induced by oxygen/glucose deprivation( OGD) and reperfusion,as well as the effects of Yinxing Mihuan oral solution on autophagy pathways mediated by endothelial nitric oxide synthase( e NOS)depended by Beclin-1. Methods: Oxygen/glucose deprivation and oxygen/glucose reintroduction model was established in murine cerebral microvascular endothelial cells and SH-SY5 Y cells by( OGD) and reperfusion. Meanwhile,different concentrations of Yinxing Mihuan Oral Solution( 5. 15 mg/m L,2. 575 mg/m L and 1. 545 mg/m L) were used,and efficient compositions were intervened. Through the observation of the concentrations of inflammatory factors IL-1 β,TNF-α and IL-6 in cell cultivated supernate and the excitation of autophagy pathways. Brain protective mechanism of Yinxing Mihuan active principles by apoptosis rate adopted the PI/Annexin V double staining analysis was discussed. Results: Yinxing Mihuan Oral Solution( 5. 15 mg/m L,2. 575 mg/m L and 1. 545 mg/m L) could restrain the contents of TNF-α,IL-1 β and IL-6 in cell culture supernatant of CMEC. Yinxing Mihuan Oral Solution( 2. 575 mg/m L) significantly promoted the phosphorylation of e NOs and Yinxing Mihuan Oral Solution( 2. 575 mg/m L and 1. 545 mg/m L) restrained the Caspase-3,LC3 II and Beclin-1 expression. The effects of L-1 β and IL-6 were significantly better than that of ginkgo biloba extract and mellea armillaria. Yinxing Mihuan Oral Solution( 2. 575 mg/m L and 1. 545 mg/m L) also decreased SH-SY5 Y cells apoptosis rate( Annexin V +/PI-cells),with significant difference. Conclusion: Yinxing Mihuan oral solution could promote the e NOs phosphorylation,which may be the interventional targets in restraining the cells' apoptosis and autophagy induced by OGD and reperfusion.
引文
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[2]韩永鹏,徐佳,朱树建.冠状动脉粥样硬化性心脏病心绞痛银杏蜜环口服溶液治疗分析[J].现代养生(下半月版),2017,32(7):69.
[3]郑咏秋,刘建勋,徐立,等.维脑康及银杏总内酯对局灶性脑缺血再灌注自噬和神经发生作用研究[J].中国中药杂志,2013,38(13):2182-2186.
[4]郑咏秋,姚明江,刘建勋,等.华佗再造浸膏对大鼠局灶性脑缺血/再灌注血脑屏障损伤的保护作用及机制研究[J].中国中药杂志,2013,38(4):585-590.
[5]Zhao J,Zhao Y,Zheng W,et al.Neuroprotective effect of curcumin on transient focal cerebral ischemia in rats[J].Brain Res,2008,1229:224-232.
[6]Zheng YQ,Liu JX,Li XZ,et al.RNA interference-mediated downregulation of Beclin1 attenuates cerebral ischemic injury in rats[J].Acta Pharmacol Sin,2009,30(7):919-927.