论文部分内容阅读
目的探讨Wnt3a在大鼠蛛网膜下腔出血(SAH)后早期脑损伤中对神经细胞自噬和凋亡的作用及影响。方法将75只健康雄性SD大鼠按随机数字表法分为假手术组(Sham组)、蛛网膜下腔出血组(SAH组)及干预组(Wnt3a组),每组25只;采用枕大池自体注血方法建立SAH模型,各组于出血后0、12、24、48、72 h取脑。Western bloting检测LC3Ⅱ与LC3Ⅰ的比值、Beclin-1、Caspase-3的表达情况,免疫组化染色技术观察各组大鼠海马的自噬和凋亡情况。结果 Western bloting结果显示,SAH组LC3Ⅱ/Ⅰ、Beclin-1出血后呈升高趋势,于24 h达高峰,24 h时间点Wnt3a组LC3Ⅱ、Beclin-1的表达较SAH组明显(P<0.05)。SAH组出血后Caspase-3升高,48h达高峰,Wnt3a组Caspase-3表达较SAH组减少(P<0.05)。免疫组化结果显示,与SAH组相比,Wnt3a组24 h时间点Beclin-1阳性神经元增多,48 h时Bax阳性神经元与凋亡细胞均减少(P<0.05)。结论 Wnt3a可以促进大鼠蛛网膜下腔出血后神经元自噬,减少神经元细胞凋亡,对神经元具有保护作用。
Objective To investigate the effect of Wnt3a on neuronal autophagy and apoptosis in early brain injury following subarachnoid hemorrhage (SAH) in rats. Methods Seventy-five healthy male Sprague-Dawley rats were randomly divided into Sham group, SAH group and Wnt3a group according to random number table. Each group consisted of 25 rats. SAH model was established by self-injection method, and the brains were harvested at 0, 12, 24, 48 and 72 h after hemorrhage in each group. Western bloting was used to detect the ratio of LC3II to LC3Ⅰ, the expression of Beclin-1 and Caspase-3, and the hippocampal autophagy and apoptosis in each group were observed by immunohistochemical staining. Results The results of Western blotting showed that the levels of LC3Ⅱ / Ⅰ and Beclin-1 in SAH group tended to increase after 24 hours and reached the peak at 24 hours. The expression of LC3Ⅱ and Beclin-1 in Wnt3a group was significantly higher than that in SAH group at 24 hours (P <0.05) . After SAH hemorrhage, Caspase-3 increased and peaked at 48h. The expression of Caspase-3 in Wnt3a group was lower than that in SAH group (P <0.05). The results of immunohistochemistry showed that the number of Beclin-1 positive neurons increased in Wnt3a group at 24 h compared with that in SAH group, and decreased in Bax positive neurons and apoptotic cells at 48 h (P <0.05). Conclusion Wnt3a can promote neuronal autophagy after subarachnoid hemorrhage in rats, reduce neuronal apoptosis and protect neurons.