目的 探索天麻素通过Notch信号通路调控BV2小胶质细胞激活的机制。 方法 体外培养BV2细胞，采用Western blot和免疫荧光双标染色检测氧糖剥夺（oxygen glucose deprivation，OGD）模型激活的BV2细胞Notch信号通路中Notch-1及下游的细胞内Notch受体结构域（intracellular Notch receptor domain，NICD）、转移因子发卡和split-1增强子（transcription factor hairy and enhancer of split-1，Hes-1）、重组结合蛋白抑制子（recombining binding protein suppressor of hairless，RBP-JK）的蛋白表达，以及天麻素干预的影响。 结果 Western blot结果显示，OGD诱导激活BV2细胞Notch信号通路相关蛋白Notch-1、NICD、Hes-1、RBP-JK的蛋白表达升高，天麻素干预降低了上述蛋白的表达。免疫荧光双标染色显示，天麻素可显著降低激活的小胶质细胞Notch-1、NICD、Hes-1和RBP-JK的表达。 结论 天麻素可通过Notch信号通路调节BV2细胞激活，从而发挥其保护作用。
Objective To investigate the mechanism of gastrodin regulating the activation of BV2 cells through Notch signal pathway. Methods BV2 cells were cultured in vitro. Western Blot and immunofluorescence double staining were used to detect the effect of gastrodin on the protein expression of Notch-1 and intracellular Notch receptor domain (NICD) , recombining binding protein suppressor of hairless (RBP-JK) and transcription factor hairy and enhancer of split-1 (Hes-1) in activated BV2 cells induced by oxygen-glucose deprivation (OGD). Results Western Blot results showed that the expression of Notch-1, NICD, Hes-1 and RBP-JK proteins was increased in OGD induced BV2 cells, and gastrodin treatment decreased the expression of these protein expressions. Double immunofluorescence staining showed that gastrodin could significantly reduce the expression of Notch-1, NICD, Hes-1 and RBP-JK proteins in activated microglia. Conclusions Gastrodin can regulate the activation of BV2 cells through Notch signal pathway to play its protective role.
[1] Van Rossum D, Hanisch UK. Microglia[J]. Metab Brain Dis, 2004, 19(3-4): 393-411. DOI: 10.1023/b:mebr.0000043984.73063.d8.
[2] 刘波, 王嘉麟, 陈师林, 等. 小胶质细胞与缺血缺氧性脑损伤关系的研究进展[J]. 中国医药导报, 2019, 16(9): 38-42. CNKI: SUN:YYCY.0.2019-09-011．
[3] 唐佳琦, 卢韵碧. 小胶质细胞与创伤性脑损伤关系的研究进展[J]. 中国细胞生物学学报, 2021, 43(8): 1714-1721. DOI: 10.11844/cjcb.2021.08.0021.
[4] Guo J, Zhang XL, Bao ZR, et al. Gastrodin regulates the Notch signaling pathway and Sirt3 in activated microglia in cerebral hypoxic-ischemia neonatal rats and in activated BV-2 microglia[J]. Neuromolecular Med, 2021, 23(3): 348-362. DOI: 10.1007/s12017-020-08627-x.
[5] 魏恩韬, 张琪. 天麻素神经保护效应的研究进展[J]. 辽宁中医杂志, 2019, 46(11): 2443-2446. DOI: 10.13192/j.issn.1000-1719.2019.11.057.
[6] 尉卫桥, 常守刚, 梁冲. 天麻素对蛛网膜下腔出血模型大鼠Sirt6/FoxO3a通路及神经功能损害的影响[J]. 卒中与神经疾病, 2021, 28(4): 415-420. DOI: 10.3969/j.issn.1007-0478.2021.04.008.
[7] 吴芳, 刘顺金, 左涵珺, 等. 天麻素对新生大鼠缺血缺氧脑损伤中小胶质细胞NOX2表达的影响[J]. 神经解剖学杂志, 2021, 37(3): 274-278. DOI: 10.16557/j.cnki.1000-7547.2021.03.004.
[8] Fernandes A, Miller-Fleming L, Pais TF. Microglia and inflammation: conspiracy, controversy or control[J]?. Cell Mol Life Sci, 2014, 71(20): 3969-3985. DOI: 10.1007/s00018-014-1670-8.
[9] 魏富芹, 黄蓉, 何海艳, 等. 天麻的药理作用及应用研究进展[J]. 中国民族民间医药, 2021, 30(11): 72-76.
[10] 鲍月宁, 曹传宇. 天麻素改善氯化钴诱导大鼠缺氧性脑神经元损伤的效果[J]. 中国医药导报, 2016, 13(14): 17-20. CNKI: SUN:YYCY.0.2016-14-004.
[11] Giaimo BD, Borggrefe T. Introduction to molecular mechanisms in notch signal transduction and disease pathogenesis[J]. Adv Exp Med Biol, 2018, 1066: 3-30. DOI: 10.1007/978-3-319-89512-3_1.
[12] 郭静, 吴芳, 袁云, 等. 天麻素对缺血缺氧脑损伤新生大鼠小胶质细胞Sirt3表达的影响[J]. 神经解剖学杂志, 2020, 36(1): 45-50. DOI: 10.16557/j.cnki.1000-7547.2020.01.008.
[13] 郭静. 天麻素对激活的小胶质细胞Notch信号通路及Sirt3表达影响的研究[D]. 昆明: 昆明医科大学, 2020. DOI: 10.27202/d.cnki.gkmyc.2020.000952.
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