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目的研究C5aR1在新生小鼠脑内及体外培养神经干细胞内的表达规律,探索C5aR1对神经干细胞增殖调节的作用。方法以免疫组化方法检测在新生小鼠脑内及体外培养的神经干细胞内C5aR1的表达;培养C5aR基因敲除小鼠来源的神经干细胞,同时利用C5aR拮抗剂处理正常小鼠来源的神经干细胞,分别观察神经干细胞的增殖状态,分析C5aR1对神经干细胞增殖的调节作用。结果 1)在新生小鼠脑内,C5aR1主要表达于海马和室下区等神经干细胞分布区域;2)C5aR1与nestin共表达于体外培养的神经干细胞内;3)C5aR特异性拮抗肽下调体外培养的神经干细胞内nestin表达并抑制其增殖;C5aR1基因敲除,神经干细胞的成球能力明显降低。结论 C5aR1在脑内以及神经干细胞内均有表达,对神经干细胞的增殖具有促进作用,其具体机制还有待进一步的实验研究。
Objective To investigate the expression of C5aR1 in neural stem cells of neonatal mice and in vitro and to explore the role of C5aR1 in the regulation of neural stem cells proliferation. Methods The expression of C5aR1 in neural stem cells cultured in vitro and in vivo was detected by immunohistochemistry. The neural stem cells derived from C5aR knockout mice were cultured, and the normal mouse-derived neural stem cells were treated with C5aR antagonist. The proliferative state of neural stem cells was observed respectively, and the regulation of C5aR1 on the proliferation of neural stem cells was analyzed. Results 1) In the newborn mouse brain, C5aR1 mainly expressed in the distribution area of neural stem cells such as the hippocampus and the subventricular zone; 2) C5aR1 and nestin co-expressed in neural stem cells cultured in vitro; 3) C5aR specific antagonist peptide down-regulated in vitro Neural stem cells nestin expression and inhibit its proliferation; C5aR1 knockout, neural stem cells into the ball ability was significantly reduced. Conclusion C5aR1 is expressed both in the brain and in neural stem cells, which may promote the proliferation of neural stem cells. The specific mechanism remains to be further studied.