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目的研究大鼠脑出血(ICH)后细胞色素c(cytc)表达和神经细胞凋亡的关系,探讨牛磺酸熊去氧胆酸(Tudca)的干预作用及其机制。方法用VII型胶原酶注入到大鼠右侧苍白球诱导ICH模型。用凋亡原位末端标记技术(TUNEL)和免疫组化方法检测血肿周围神经细胞凋亡和cytc表达的动态变化。透射电镜观察术后3d血肿周围神经元超微结构的变化。结果:ICH后6h cytc表达开始增加,1d达高峰,此后逐渐减少,7d时回落至假手术组水平。ICH后6h血肿周边组织可见TUNEL阳性细胞,3d达高峰,然后逐渐减少,7d、10d时仍见较多凋亡细胞。ICH后7d内cytc表达和神经细胞凋亡呈正相关(r=0.6357,P<0.01)。Tudca治疗组cytc和TUNEL阳性细胞数较模型组明显减少(P<0.05~0.01),神经元超微结构改变亦比模型组减轻。结论ICH后血肿周围神经细胞损伤有凋亡机制参与,cytc释放是神经细胞凋亡的一个关键事件,Tudca可以有效减轻脑出血后的细胞凋亡,其机制之一是抑制凋亡通路cytc的释放。
Objective To study the relationship between cyt c expression and neuronal apoptosis after intracerebral hemorrhage (ICH) in rats and the possible mechanism of Tudca. Methods The type IV collagenase was injected into the right globus pallidus in rats to induce ICH model. TUNEL and immunohistochemistry were used to detect the dynamic changes of apoptosis and cyt c expression in the perihematomal cells. TEM observation of ultrastructural changes of neurons around hematoma after 3 days. Results: Six hours after ICH, the expression of cytc began to increase and peaked on day 1, then decreased gradually and dropped to the level of sham operation group on the 7th day. TUNEL positive cells were observed in the perihematoma tissue around the hematoma 6h after ICH, reaching the peak on 3d and then gradually decreasing, and more apoptotic cells were still observed on the 7th and 10th day. There was a positive correlation between cyt c expression and neuronal apoptosis within 7 days after ICH (r = 0.6357, P <0.01). Compared with the model group, the number of cyt c and TUNEL positive cells in the Tudca treatment group was significantly decreased (P <0.05-0.01), and ultrastructural changes in neurons were also alleviated compared with the model group. Conclusions After ICH, there are apoptotic mechanisms involved in the injury of peripheral nerve cells. Cytotoxicity of cyt c is a key event in neuronal apoptosis. Tudca can effectively reduce apoptosis after intracerebral hemorrhage. One of the mechanisms is to inhibit the release of the apoptotic pathway cytc .