论文部分内容阅读
为了研究体外培养大鼠胚胎脊髓神经细胞损伤前后的凋亡变化及caspase-3的表达情况,本实验建立了一个模拟脊髓横断损伤后脊髓组分-原代培养的脊髓神经细胞机械损伤模型,并用Hoechst33342/PI双染法检测脊髓神经细胞的凋亡变化,用免疫荧光染色方法检测caspase-3的表达。结果显示:(1)损伤前几乎未见凋亡的神经细胞,损伤后6 h凋亡细胞开始出现,损伤后12 h明显增多,1 d时达高峰,但损伤后3 d凋亡细胞开始呈明显下降的趋势,至损伤后7 d又开始增加,一直持续到损伤后14d;(2)相应时刻caspase-3表达趋势的改变与脊髓神经细胞凋亡趋势的变化基本相同;(3)经过caspase-3抑制剂Ac-DEVD-CHO干预后,凋亡细胞的数量与caspase-3的表达均减少,并呈剂量依赖性。以上结果提示,机械损伤可以诱发体外培养的脊髓神经细胞凋亡,且此凋亡的发生可能与caspase-3的表达有关。
In order to study the changes of apoptosis and the expression of caspase-3 in cultured rat embryonic spinal cord neurons before and after injury, we established a model of spinal cord injury-induced spinal neuronal cell injury following spinal cord transection Hoechst33342 / PI double staining was used to detect the apoptosis of spinal cord neurons. The expression of caspase-3 was detected by immunofluorescence staining. The results showed that: (1) There were almost no apoptotic neurons before injury, and apoptotic cells began to appear at 6 h after injury. The apoptotic cells began to increase 12 h after injury and peaked at d 1, but apoptotic cells began to show on day 3 after injury (2) At the corresponding time, the change of caspase-3 expression was basically the same as that of changes of apoptosis of spinal cord nerve cells; (3) After caspase -3 inhibitor Ac-DEVD-CHO intervention, the number of apoptotic cells and the expression of caspase-3 were reduced and in a dose-dependent manner. The above results suggest that mechanical injury can induce apoptosis of spinal cord nerve cells cultured in vitro, and the occurrence of this apoptosis may be related to the expression of caspase-3.