Neuroprotective effect of gastrodin in methamphetamine-induced apoptosis through regulating cAMP/PKA/CREB pathway in cortical neuron
Neuroprotective effect of gastrodin in methamphetamine-induced apoptosis through regulating cAMP/PKA/CREB pathway in cortical neuron
复制标题
天麻素通过调节皮质神经元cAMP/PKA/CREB通路对甲基苯丙胺诱导的细胞凋亡的神经保护作用
DOI：
10.1177/0960327120911438
复制
发表时间：
2020-03-12
影响因子：
2.8
通讯作者：
Hong, S-J
中科院分区：
文献类型：
--
作者：
Ma, C-L;Li, L.;Hong, S-J
目的：甲基苯丙胺（Methamphetamine，MA）滥用可引起神经毒性和神经细胞凋亡。天麻素是一种传统的中草药，用于治疗神经损伤、脊髓损伤以及一些中枢神经系统疾病。方法：原代培养的大鼠大脑皮层神经元分为4组（对照组、MA组、MA +天麻素组、MA+天麻素+小干扰RNA组）。采用末端脱氧核苷酸转移酶缺口末端标记法检测凋亡细胞，CCK-8法检测细胞存活率，免疫荧光法检测Tuj 1阳性细胞数和平均轴突长度，并检测环磷酸腺苷（cAMP）、蛋白激酶A（PKA）、cAMP反应元件结合（CREB）、结果：CCK-8法显示0.5mM MA是诱导神经毒性的最适浓度（p < 0.01）。25 mg/L天麻素预处理对神经细胞的保护作用最大。MA组cAMP、PKA、磷酸化PKA、CREB、磷酸化CREB和BDNF蛋白表达水平降低，天麻素预处理组上调这些蛋白的表达水平（p < 0.01）。对照组、MA组和天麻素组PKA和CREB蛋白表达无明显变化。与MA+天麻素组相比，MA +天麻素+ small interfering RNA组Tuj 1阳性细胞数和平均轴突长度明显减少，凋亡细胞数明显增加（p <0. 05）。结论：天麻素对MA诱导的神经毒性具有神经保护作用，其作用机制可能与调节cAMP/PKA/CREB信号通路和上调BDNF表达有关。
Objective:Methamphetamine (MA) abuse induces neurotoxicity and causes neuronal cell apoptosis. Gastrodin is a traditional Chinese herbal medicine used for the treatment of nerve injuries, spinal cord injuries, and some central nervous system diseases as well. The present study investigated the neuroprotective effects of gastrodin against MA-induced neurotoxicity in neuronal cells and its potential protective mechanism.Methods:The primary cortex neuronal culture was divided into four groups (control group, MA group, MA + gastrodin group, and MA + gastrodin + small interfering RNA group). The neurotoxicity of MA was assessed by detecting apoptotic cells by terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay and cell viability by cell counting kit 8 (CCK-8) method, the Tuj1-positive cells and the average axonal length were detected by immunofluorescence, and the expressions of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP-response element-binding (CREB), and brain-derived neurotrophic factor (BDNF) proteins were detected by Western blot.Results:The results of CCK-8 assay showed that 0.5 mM MA was an optimal concentration that induced neurotoxicity (p < 0.01). Pretreatment with 25 mg/L gastrodin exerted maximum protective effects on neuronal cells. The expression levels of cAMP, PKA, phosphorylated PKA, CREB, phosphorylated CREB, and BDNF proteins were decreased in the MA group, and pretreatment with gastrodin upregulated the expression levels of these proteins (p < 0.01). The expressions of PKA and CREB proteins showed no significant changes in the control group, MA group, and gastrodin group. Compared the MA + gastrodin + small interfering RNA group with MA + gastrodin group, the Tuj1-positive cells and the average axonal length were decreased significantly, while the number of apoptotic cells was increased (p < 0.05).Conclusion:Gastrodin has neuroprotective effects against MA-induced neurotoxicity, which exerts neuroprotective effects via regulation of cAMP/PKA/CREB signaling pathway and upregulates the expression of BDNF.