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切除成年大鼠大脑双侧顶叶部分皮质,在术后不同时间取损伤周围脑组织,制成脑创伤组织提取液(BWTE);并建立新生大鼠大脑皮质神经元体外培养模型,检测BWTE中的神经元营养因子(NTFs)和促神经突起生长因子(NPFs)。为探讨这些因子的来源,是否与脑创伤区早期出现大量的巨噬细胞有关,我们培养巨噬细胞,收集巨噬细胞条件性培养基(MφCM),检测MφCM对大脑皮质神经元的神经营养活性。此外,亦观察BWTE和MφCM对PC 12细胞的作用,以进一步探讨NPFs的作用机制。实验结果表明,BWTE和MφCM中含有对大脑皮质神经元的NTFs和NPFs。这些因子于脑损伤后第4天开始出现,第5天达高峰(NPFs在第9天还出现一个高峰),第13天仍有活性。MφCM的NTF活性不及BWTE,但NPF活性则比BWTE强。BWTE和MφCM对PC 12细胞也具有NPF活性。通过实验分析认为,BWTE中的神经营养因子,早期主要来源于巨噬细胞,后期可能与星形胶质细胞有关。这些因子成分复杂,可能含多种有效成分,有待今后进一步探讨。
The parietal cortex of bilateral parietal lobe of adult rats was excised and the injured brain tissue was harvested at different times after operation to make brain trauma tissue extract (BWTE). The model of cultured cortical neurons in neonatal rats was established and cultured in BWTE Of neuronal nutrition factors (NTFs) and pro-neurite outgrowth factors (NPFs). To investigate whether the origin of these factors is related to the early appearance of large numbers of macrophages in brain trauma areas, we cultured macrophages and collected macrophage conditioned medium (MφCM) to detect the neurotrophic activity of MφCM on neurons of cerebral cortex . In addition, the effects of BWTE and MφCM on PC 12 cells were also observed to further explore the mechanism of action of NPFs. Experimental results show that BWTE and MφCM contain NTFs and NPFs on cerebral cortex neurons. These factors began to appear on the fourth day after brain injury, peaked on the fifth day (NPFs peaked on the ninth day), and remained active on the thirteenth day. NTF activity of MφCM was lower than that of BWTE, but NPF activity was stronger than that of BWTE. BWTE and MφCM also have NPF activity on PC 12 cells. Through experimental analysis, neurotrophic factors in BWTE mainly come from macrophages in the early stage and may be related to astrocytes in the late stage. The complex composition of these factors, may contain a variety of active ingredients, to be further explored in the future.