Department of Neurology, Hunan Provincial Brain Hospital, Changsha, China.
Hum Exp Toxicol. 2021 Sep;40(9):1422-1433. doi: 10.1177/0960327121996021. Epub 2021 Mar 4.
Cerebral ischemia is the main cause of permanent adult disabilities worldwide. This study investigated the reparative effects and potential mechanisms of methylphenidate (MPH), a medication for the treatment of attention-deficit/hyperactivity disorder.
In vitro oxygen-glucose deprivation/reperfusion (OGD/R) and in vivo cerebral ischemia-reperfusion models were established. Sprague-Dawley (SD) rats were randomly divided into four groups ( = 20): Sham, Model, and MPH (0.5 and 1 mg/kg). Rats in MPH groups were treated with 0.5 or 1 mg/kg MPH via intraperitoneal injection for 7 days. Rats in the Sham and Model groups were treated with PBS during the same period. Cell viability was measured using MTT assay. Apoptosis was detected by Annexin V/PI staining. Protein expression was detected by Western blot. The volume of cerebral infarction was detected by triphenyltetrazolium chloride (TTC) staining. The DNA damage in ischemic brain tissues was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.
MPH treatment significantly reduced OGD/R-induced cell damage, shown by the increased cell viability and decreased apoptotic rate. p-AMPK and p-ACC protein expression increased in the OGD/R model after MPH treatment. The addition of AMPK inhibitor largely abolished the neuroprotective effects of MPH, evidenced by the reduced cell viability, increased apoptotic rate, and decreased protein expression of p-AMPK as well as p-ACC. Moreover, MPH treatment significantly alleviated the cerebral ischemia-reperfusion injury and decreased apoptosis in brain tissues, which may be associated with the AMPK/ACC pathway.
MPH exerted protective activities against oxidative stress in the OGD/R model and ameliorated brain damage of rats in the middle cerebral artery occlusion model, at least in part, through activating the AMPK pathway. These data demonstrated neuroprotective properties of MPH and highlighted it as a potential therapeutic agent against cerebral ischemia-reperfusion injury.
脑缺血是全球成年人永久性残疾的主要原因。本研究探讨了哌醋甲酯（MPH）对治疗注意缺陷多动障碍的治疗效果及其潜在机制。
体外氧葡萄糖剥夺/再灌注（OGD/R）和体内脑缺血再灌注模型建立。将 Sprague-Dawley（SD）大鼠随机分为四组（每组 20 只）：假手术组、模型组、MPH（0.5 和 1mg/kg）组。MPH 组大鼠通过腹腔注射给予 0.5 或 1mg/kg MPH 治疗 7 天。假手术组和模型组大鼠同期给予 PBS 处理。采用 MTT 法检测细胞活力。采用 Annexin V/PI 染色检测细胞凋亡。采用 Western blot 检测蛋白表达。采用氯化三苯基四氮唑（TTC）染色检测脑梗死体积。采用末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）法检测缺血脑组织中的 DNA 损伤。
MPH 治疗可显著减轻 OGD/R 诱导的细胞损伤，表现为细胞活力增加和凋亡率降低。OGD/R 模型中 MPH 处理后 p-AMPK 和 p-ACC 蛋白表达增加。加入 AMPK 抑制剂后，MPH 的神经保护作用明显减弱，表现为细胞活力降低、凋亡率增加、p-AMPK 和 p-ACC 蛋白表达减少。此外，MPH 治疗可显著减轻大脑中动脉闭塞模型大鼠的脑缺血再灌注损伤和脑组织凋亡，这可能与 AMPK/ACC 通路有关。
MPH 对 OGD/R 模型中的氧化应激具有保护作用，并通过激活 AMPK 通路改善大脑中动脉闭塞模型大鼠的脑损伤，至少部分是通过激活 AMPK 通路。这些数据表明 MPH 具有神经保护作用，可将其作为治疗脑缺血再灌注损伤的潜在治疗药物。
