Beagle犬含药脑脊液干预皮层神经元出血损伤细胞凋亡时间窗的实验研究

The Protection Effect of Beagle Dog Cerebrospinal Fluid on the Neuron

目的探讨中药复方DSQ-03对Beagle犬含药脑脊液体外培养的皮层神经元凝血酶合并缺血缺氧损伤的保护作用,并探讨其作用的时间窗,为进一步开展临床研究提供依据;方法采用脑脊液药理学和原代神经细胞培养的方法,用Beagle犬的含DSQ-03脑脊液体外培养SD大鼠大脑皮质的神经细胞。用凝血酶合并缺血缺氧(TIH)损伤神经细胞,建立急性脑出血(AICH)的体外细胞模型。在TIH损伤后6h、12h、24h、72h,用流式细胞仪检测细胞凋亡率。结果AICH细胞模型复制后,神经细胞凋亡在0h～72h都有动态变化;Beagle犬含药脑脊液干预ICH的细胞凋亡时间窗:0h～24h是最佳干预时机,0h～72h内仍有意义。结论DSQ-03含药脑脊液可抑制神经细胞凋亡,其最佳时间窗为0h～24h。

Objective To observe the therapeutic time window of acute intracerebral hemorrhage （AICH） intervened by Beagle dog cerebrospinal fluid containing DSQ- 03 herbal medicine. Methods ① The cerebral cortex neuron of SD rats were cultured by Beagle dog CSF containing the DSQ-03 in vitro.② The AICH cell models were made in vitro. ③ Following injury of TIH 6 h, 12 h,24 h,48 h,72 h, the apoptosis of the neuron after AICH were measured by flow cytometry. Results From 0 h to 72 h after AICH, the apoptosis of the neuron appeared. The optimal effect of inhibiting the neural apoptosis was at 0 h to 24 h, and still persisted from 0 h to 72 h. Conclusion Beagle dog CSF containing DSQ- 03 herbal composition was able to inhibit the neural apoptosis. The therapeutic time window of AICH treated with DSQ- 03 herbal medicine was 0 h to 24 h.