| 引用本文:郑咏秋1，张业昊1，刘建勋1，徐立1，李磊1，王益民2，王勇2，王光蕊1.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[J].世界中医药,2018,(01):. |
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| 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 |
| Study on Protective Effects and Mechanisms of Yinxing Mihuan Oral Solution on Cerebral Microvascular Endothelial Cells and Inflammatory Injury of SH-SY5Y Cells Induced by Oxygen/Glucose Deprivation and Reperfusion |
| 投稿时间：2017-12-20 |
| DOI：10.3969/j.issn.1673-7202.2018.01.003 |
| 中文关键词: 银杏蜜环 缺氧缺糖再灌注 脑微血管内皮细胞 SH-SY5Y 自噬 |
| English Keywords:Yinxing Mihuan Oral Solution Oxygen/glucose deprivation and reperfusion Cerebral microvascular endothelial cells SH-SY5Y Autophagy |
| 基金项目:国家重点基础研究发展计划(973计划)项目(2015CB554405);国家自然科学基金项目(81374036);中国中医科学院院内联合创新专项(ZZ11-026) |
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| 中文摘要: |
| 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eNOs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/mL、2.575 mg/mL和1.545 mg/mL)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/mL、2.575 mg/mL和1.545 mg/mL3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/mL和1.545 mg/mL可抑制Caspase3、LC3II和Beclin-1表达。同时,银杏蜜环口服溶液2.575 mg/mL明显促进eNOs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/mL和1.545 mg/mL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eNOs的磷酸化来抑制缺糖缺氧再灌引发的细胞凋亡和自噬。 |
| English Summary: |
| To study the protective effects and mechanisms of Yinxing Mihuan Oral Solution for cerebral microvascular endothelial cells (CMEC) and SH-SY5Y 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 (eNOS) depended by Beclin-1. Methods: Oxygen/glucose deprivation and oxygen/glucose reintroduction model was established in murine cerebral microvascular endothelial cells and SH-SY5Y cells by (OGD) and reperfusion. Meanwhile, different concentrations of Yinxing Mihuan Oral Solution (5.15 mg/mL, 2.575 mg/mL and 1.545 mg/mL) were used, and efficient compositions were intervened. Through the observation of the concentrations of inflammatory factors IL-1β, 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/mL, 2.575 mg/mL and 1.545 mg/mL) could restrain the contents of TNF-α, IL-1β and IL-6 in cell culture supernatant of CMEC. Yinxing Mihuan Oral Solution (2.575 mg/mL) significantly promoted the phosphorylation of eNOs and Yinxing Mihuan Oral Solution (2.575 mg/mL and 1.545 mg/mL) restrained the Caspase-3, LC3II and Beclin-1 expression. The effects of L-1β and IL-6 were significantly better than that of ginkgo biloba extract and mellea armillaria. Yinxing Mihuan Oral Solution (2.575 mg/mL and 1.545 mg/mL) also decreased SH-SY5Y cells apoptosis rate (Annexin V+/PI-cells), with significant difference. Conclusion: Yinxing Mihuan oral solution could promote the eNOs phosphorylation, which may be the interventional targets in restraining the cells' apoptosis and autophagy induced by OGD and reperfusion. |
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