Park Won Soon, Sung Dong Kyung, Kang Saem, Koo Soo Hyun, Kim Yu Jin, Lee Jang Hoon, Chang Yun Sil, Lee Munhyang
Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Korea.
J Korean Med Sci. 2006 Apr;21(2):337-41. doi: 10.3346/jkms.2006.21.2.337.
This study was done to determine the neuroprotective effect of cycloheximide on neonatal hypoxic-ischemic brain injury. Seven day-old newborn rat pups were subjected to 90 min of 8% oxygen following a unilateral carotid artery ligation. The extent of cerebral infarction was evaluated at 1 and 4 week of recovery. Apoptosis was identified by performing terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining and flow cytometry with a combination of fluoresceinated annexin V and propidium iodide. Brain infarction area was significantly increased at 4 week compared to 1 week after hypoxia-ischemia in the control group. With cycloheximide treatment, the number of TUNEL positive cells in the ipsilateral cerebral cortex at 48 hr and peri-infarct area at 1 and 4 week of recovery was significantly reduced, both apoptotic and necrotic cells by flow cytometry 48 hr after the injury were significantly reduced, and the extent of cerebral infarction at 1 and 4 week of recovery was also significantly attenuated compared to the hypoxia-ischemia control group. In summary, our data suggest that apoptosis plays an important role in the development of delayed infarction, and inhibition of apoptosis with cycloheximide significantly reduces the ensuing cerebral infarction in a newborn rat pup model of cerebral hypoxia-ischemia.
本研究旨在确定放线菌酮对新生儿缺氧缺血性脑损伤的神经保护作用。将7日龄新生大鼠幼崽在单侧颈动脉结扎后置于8%氧气环境中90分钟。在恢复1周和4周时评估脑梗死范围。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)染色以及结合荧光素化膜联蛋白V和碘化丙啶的流式细胞术来鉴定细胞凋亡。与缺氧缺血后1周相比,对照组在4周时脑梗死面积显著增加。放线菌酮治疗后,恢复48小时时同侧大脑皮质TUNEL阳性细胞数量以及恢复1周和4周时梗死周边区域的TUNEL阳性细胞数量均显著减少,损伤后48小时通过流式细胞术检测的凋亡细胞和坏死细胞数量均显著减少,并且与缺氧缺血对照组相比,恢复1周和4周时脑梗死范围也显著减轻。总之,我们的数据表明细胞凋亡在迟发性梗死的发生发展中起重要作用,并且在新生大鼠脑缺氧缺血模型中,用放线菌酮抑制细胞凋亡可显著减少随后发生的脑梗死。