异氟醚诱导神经元损伤中 P2YR12 的失活通过改变 TLR-4/BDNF/TNF-α 起作用。
Inactivation of P2YR12 contributes to isoflurane-induced neuronal injury by altering TLR-4/BDNF/TNF-α.
机构信息
Department of Anesthesiology, Weifang People's Hospital, Weifang, Shandong, China.
Department of Operating Theatre, Weifang Seventh People's Hospital, Weifang, Shandong, China.
出版信息
Folia Neuropathol. 2019;57(2):161-169. doi: 10.5114/fn.2019.86295.
The present investigation evaluated the effect of inhibiting the P2Y12 gene on anaesthetic-induced neuronal injury in a rat model. Neuronal injury was induced by exposing the animals for 6 h to 30% oxygen containing 0.75% isoflurane and 1.2 mg/kg prasugrel (a P2Y12 inhibitor) p.o. for 14 days. Cognitive function was determined by the Morris water maze, and the neurological severity score was determined. Enzyme-linked immunosorbent assay was used to estimate the level of oxidative stress and mediators of inflammation in brain tissues of isoflurane-induced neuronal injury rats. Apoptosis of neuronal cells was estimated by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) and western blot assays. Real time-polymerase chain reaction was performed to estimate the expression levels of several proteins. The data revealed that inhibiting the P2Y12 gene ameliorated changes in the modified neurological severity score and cognitive function in neuronal injury rats. Moreover the levels of proinflammatory mediators, oxidative stress, and cyclic AMP, and the number of TUNEL-positive cells, decreased significantly (p < 0.01) in the prasugrel-treated group compared to the negative control group. In addition, apoptosis of neuronal cells decreased in the prasugrel-treated group, as it ameliorated expression of the PI3K, Bcl-2, Bad, and Akt proteins in the isoflurane-induced neuronal injury rats. Expression of brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) proteins was enhanced, whereas the Toll-like receptor-4 (TLR-4) and nuclear factor κB (NF-κB) proteins decreased in the brain tissues of the prasugrel-treated group compared to the negative control group of rats. These results suggest that inhibiting the P2YR12 gene protects against neuronal injury in isoflurane-induced neuronal injury rats. Inhibiting the P2YR12 gene ameliorated neuronal apoptosis by regulating the BDNF/TLR-4/TNF-α pathway.
本研究评价了抑制 P2Y12 基因对大鼠模型中麻醉诱导神经元损伤的影响。通过使动物暴露于含有 0.75%异氟烷和 1.2mg/kg普拉格雷（P2Y12 抑制剂）的 30%氧气中 6 小时，并用普拉格雷灌胃 14 天来诱导神经元损伤。通过 Morris 水迷宫确定认知功能，并用神经严重程度评分确定。通过酶联免疫吸附试验估计异氟烷诱导的神经元损伤大鼠脑组织中氧化应激和炎症介质的水平。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）和蛋白质印迹法评估神经元细胞凋亡。通过实时聚合酶链反应估计几种蛋白质的表达水平。数据显示，抑制 P2Y12 基因可改善神经元损伤大鼠改良神经严重程度评分和认知功能的变化。此外，与阴性对照组相比，普拉格雷治疗组促炎介质、氧化应激和环磷酸腺苷的水平以及 TUNEL 阳性细胞的数量显著降低（p<0.01）。此外，神经元细胞凋亡减少，因为它改善了异氟烷诱导的神经元损伤大鼠中 PI3K、Bcl-2、Bad 和 Akt 蛋白的表达。与阴性对照组相比，普拉格雷治疗组脑源性神经营养因子（BDNF）和原肌球蛋白受体激酶 B（TrkB）蛋白的表达增强，而 Toll 样受体-4（TLR-4）和核因子 κB（NF-κB）蛋白的表达降低。这些结果表明，抑制 P2YR12 基因可防止异氟烷诱导的神经元损伤大鼠的神经元损伤。抑制 P2YR12 基因通过调节 BDNF/TLR-4/TNF-α 通路改善神经元凋亡。
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