江沂,刘燕娣,刘婉,陈海敏
JIANG Yi,LIU Yandi,LIU Wan,CHEN Haimin

• 1:广州医科大学附属中医医院病理科

摘要(Abstract)：

目的 研究天麻素（GAS）对神经母细胞瘤（SH-SY5Y）细胞突触相关蛋白表达的影响及其神经保护作用。方法 采用CCK8法筛选GAS安全浓度，GAS单独或联合Aβ_(1-42)处理SH-SY5Y细胞后，采用实时荧光定量PCR及蛋白质印迹法（Western blot）分别测定突触相关蛋白突触素（SYN）、发动蛋白1(DYN1)、突触后致密蛋白95(PSD95) mRNA及蛋白表达水平。结果 CCK8结果显示在0～150μmol/L GAS时细胞活力无明显变化，可作为GAS处理细胞的安全浓度；0～120μmol/L GAS处理SH-SY5Y细胞后，SYN、DYN1及PSD95 mRNA及蛋白水平均上调、并随着GAS浓度的升高而升高，差异有统计学意义（P<0.05）；与对照组比较，Aβ_(1-42)组细胞SYN、DYN1、PSD95 mRNA及蛋白表达水平明显降低，差异有统计学意义（P<0.01）；与Aβ组比较，Aβ+GAS组细胞SYN、DYN1、PSD95 mRNA及蛋白表达水平明显上调，差异有统计学意义（P<0.01）。结论 GAS可能通过上调细胞突触相关蛋白表达来发挥神经保护作用。
Objective To study the effect of gastrodin(GAS) on the expression of synapse-related proteins and its neuroprotective effects in neuroblastoma( SH-SY5 Y) cells.Methods GAS safety concentrations were screened by CCK8, and after treating SH-SY5 Y cells with GAS alone or in combination with Aβ_(1-42), synapse-associated protein synaptin(SYN), initiator protein 1(DYN1), the mRNA and protein expression levels of postsynaptic compact protein 95(PSD95) were determined by quantitative real-time PCR and Western blot, respectively.Results The CCK8 results showed no significant change in cell viability at 0 to 150 μmol/L GAS and served as a safe concentration in cells treated with GAS. After treating SH-SY5 Y cells with 0-120 μmol/L GAS, SYN, DYN1, and PSD95 mRNA and protein levels were increased and increased with increasing GAS concentration, and the difference was statistically significant(P< 0. 05). Compared with the control group, the SYN, DYN1,PSD95 mRNA, and protein expression levels were significantly decreased in the Aβ_(1-42) group, and the difference was statistically significant(P< 0. 01). Compared with group Aβ, the SYN, DYN1, PSD95 mRNA and protein expression levels were significantly upregulated in the Aβ + GAS group, and the difference was statistically significant(P< 0. 01).Conclusion GAS may exert a neuroprotective effect by upregulating the protein expression of cellular synapse-related.

关键词(KeyWords)： 天麻素;β淀粉样蛋白;SH-SY5Y细胞;突触素;发动蛋白1;突触后致密蛋白95
gastrodin (GAS);β-amyloid protein (Aβ);SH-SY5Y cell;synaptophin (SYN);initiator protein 1 (DYN1);postsynaptic compact protein 95 (PSD95)

Abstract:

Keywords:

基金项目(Foundation): 广东省卫生健康委基金项目（B2021158）

作者(Author): 江沂,刘燕娣,刘婉,陈海敏
JIANG Yi,LIU Yandi,LIU Wan,CHEN Haimin

DOI: 10.19367/j.cnki.2096-8388.2022.03.007

参考文献(References)：

• [1] DETURE M A,DICKSON D W. The neuropathological diagnosis of Alzheimer's disease[J]. Mol Neurodegener,2019, 14(1):32.
• [2] CHEN Y, FU A K Y, IP N Y. Synaptic dysfunction in Alzheimer's disease:Mechanisms and therapeutic strategies[J]. Pharmacol Ther,2019, 195:186-198.
• [3] YANG T, ZHU Z, YIN E, et al. Alleviation of symptoms of Alzheimer's disease by diminishing Aβ neurotoxicity and neuroinflammation[J]. Chem Sci, 2019, 10(43):10149-10158.
• [4] SHENG M, SABATINI B L, SUDHOF T C. Synapses and Alzheimer's disease[J]. Cold Spring Harb Perspect Biol, 2012, 4(5):747-749.
• [5] LIU Y, GAO J, PENG M, et al. A review on central nervous system effects of gastrodin[J]. Front Pharmacol,2018, 9:24.
• [6] HAYASHI J, SEKINE T, DEGUCHI S, et al. Phenolic compounds from Gastrodia rhizome and relaxant eects of related compounds on isolated smooth muscle preparation[J]. Phytochemistry,2002, 59:513-519.
• [7] CACCAMO A, ODDO S, BILLINGS L M, et al. M1 receptors play a central role in modulating AD-like pathology in transgenic mice[J]. Neuron,2006,2(3):230-230.
• [8]王晓玲，吕园园，张淑丽，等．SH-SY5Y细胞α7神经型尼古丁受体基因沉默对突触相关蛋白的影响[J]．中风与神经疾病杂志．2016, 33(12):1080-1083.
• [9] PINHEIRO L, FAUSTINO C. Therapeutic strategies targeting amyloid-beta in Alzheimer's disease[J]. Curr Alzheimer Res, 2019, 16(5):418-452.
• [10]LANE C A, HARDY J, SCHOTT J M. Alzheimer's disease[J]. Eur J Neurol,2018, 25(1):59-70.
• [11]JU Y, TAM K Y. Pathological mechanisms and therapeutic strategies for Alzheimer's disease[J]. Neural Regeneration Research, 2022, 17(3):7.
• [12]陈书仪，孙洁，郭爱，等．突触可塑性与阿尔茨海默病[J]．生命的化学，2017(4):143-147.
• [13]KARISETTY B C, BHATNAGAR A, ARMOUR E M, et al. Amyloid-β peptide impact on synaptic function and neuroepigenetic gene control reveal new therapeutic strategies for Alzheimer's disease[J]. Front Mol Neurosci,2020, 13:577622.
• [14]YANG T, ZHU Z, YIN E, et al. Alleviation of symptoms of Alzheimer's disease by diminishing Aβ neurotoxicity and neuroinflammation[J]. Chem Sci, 2019, 10(43):10149-10158.
• [15] ELHUSSEIN A E, SCHNELL E, CHETKOVICH D M,et al. PSD-95 involvement in maturation of excitatory synapses. Science, 2000, 290(5495):1364-1368.
• [16]WANG X L, DENG Y X, GAO Y M, et al. Activation ofα7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synapticassociated proteins and the CaM-CaMKII-CREB signaling pathway[J]. Aging(Albany NY),2020, 12(1):543-570.
• [17]梁坚，赵鹏，李彬，等．天麻软胶囊的毒性研究[J]．中国热带医学，2005, 5(9):1986-1988.
• [18]张勇，席刚明，周少华．天麻及其成分对神经系统的作用[J]．国际中医中药杂志，2006, 28(5):268-271.
• [19]HU Y, LI C, SHEN W. Gastrodin alleviates memory deficits and reduces neuropathology in a mouse model of Alzheimer's disease[J]. Neuropathology, 2014, 34(4):370-377.
• [20]ZHANG H, YUAN B, HUANG H, et al. Gastrodin induced HO-1 and Nrf2 up-regulation to alleviate H2O2-induced oxidative stress in mouse liver sinusoidal endothelial cells through p38 MAPK phosphorylation[J]. Braz J Med Biol Res,2018, 51(10):e7439.

文章评论(Comment)：