To demonstrate the role of the rate-limiting and ATP-dependent gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PCK) in oxidative and lactic stress and the effect of phenothiazine on PCK after stroke, a total of 168 adult male Sprague Dawley rats (3 months old, 280–300 g) underwent 2-h intraluminal middle cerebral artery occlusion (MCAO) and reperfusion for 6, 24, 48 h, or 7 days. Phenothiazine (chlorpromazine and promethazine (C+P)) (8 mg/kg) and 3-mercaptopicolinic acid (3-MPA, a PCK inhibitor, 100 μM) were administered at reperfusion onset. The effects of phosphoenolpyruvate, 3-MPA, or PCK knockdown were studied in neuronal cultures subjected to oxygen/glucose deprivation. Reactive oxygen species, lactate, phosphoenolpyruvate (PEP; a gluconeogenic product), mRNA, and protein of total PCK, PCK-1, and PCK-2 increased after MCAO and oxygen–glucose deprivation (OGD). Oxaloacetate (a gluconeogenic substrate) decreased, while PEP and glucose were increased, suggesting reactive gluconeogenesis. These changes were attenuated by phenothiazine, 3-MPA, or PCK shRNA. PCK-1 and -2 existed primarily in neurons, while the effects of ischemic stroke on the PCK expression were seen predominately in astrocytes. Thus, phenothiazine reduced infarction and oxidative/lactic stress by inhibiting PCKs, leading to functional recovery.

为了证明限速和 ATP 依赖性糖异生酶磷酸烯醇丙酮酸羧激酶 (PCK) 在氧化和乳酸应激中的作用以及吩噻嗪对中风后 PCK 的影响，总共 168 只成年雄性 Sprague Dawley 大鼠（3 个月大，280 –300 g) 进行了 2 小时腔内大脑中动脉闭塞 (MCAO) 和再灌注 6、24、48 小时或 7 天。在再灌注开始时给予吩噻嗪（氯丙嗪和异丙嗪 (C+P)）（8 mg/kg）和 3-巯基丙氨酸（3-MPA，一种 PCK 抑制剂，100 μM）。在缺乏氧气/葡萄糖的神经元培养物中研究了磷酸烯醇式丙酮酸、3-MPA 或 PCK 敲低的影响。活性氧、乳酸、磷酸烯醇式丙酮酸（PEP；一种糖异生产物）、mRNA 和总 PCK、PCK-1 的蛋白质、和 PCK-2 在 MCAO 和氧-葡萄糖剥夺（OGD）后增加。草酰乙酸（一种糖异生底物）减少，而 PEP 和葡萄糖增加，表明反应性糖异生。这些变化被吩噻嗪、3-MPA 或 PCK shRNA 减弱。PCK-1和-2主要存在于神经元中，而缺血性卒中对PCK表达的影响主要见于星形胶质细胞。因此，吩噻嗪通过抑制 PCK 来减少梗塞和氧化/乳酸应激，从而导致功能恢复。而缺血性卒中对 PCK 表达的影响主要见于星形胶质细胞。因此，吩噻嗪通过抑制 PCK 来减少梗塞和氧化/乳酸应激，从而导致功能恢复。而缺血性卒中对 PCK 表达的影响主要见于星形胶质细胞。因此，吩噻嗪通过抑制 PCK 来减少梗塞和氧化/乳酸应激，从而导致功能恢复。