环磷酰胺荷瘤小鼠numb-Notch信号途径关键基因筛选及针灸对其影响的研究

Research on key genes screening in numb-Notch signal pathway of cyclophosphamide tumor-bearing mice and effects of acupuncture and moxibustion

目的:筛选并验证针灸与环磷酰胺(CTX)荷瘤小鼠骨髓造血细胞相关的关键基因,从numb-Notch信号途径方面探讨针灸改善化疗后骨髓抑制的作用机制。方法:以CTX分别造成健康体质小鼠、荷瘤体质小鼠骨髓抑制模型,小通量基因筛选、免疫组化验证各组小鼠骨髓numb-Notch信号途径上显著性差异基因,分析针灸对CTX荷瘤小鼠骨髓造血细胞中关键基因的影响。结果:numb1、numb2、Notch2、jag1属于小鼠骨髓造血细胞Notch信号途径相关基因,且具有显著性差异,为关键基因;针刺、艾灸可上调numb1、numb2蛋白表达水平,下调Notch2、jag1蛋白表达水平。结论:上调numb-Notch信号途径显著差异基因numb1、numb2的表达,进而下调Notch2、jag1的表达,抑制过度激活的Notch信号途径,增强骨髓造血功能,可能是针灸减轻CTX化疗引起的骨髓抑制、改善造血功能的微观机制之一。

Objective:Screening and verifying the key genes related to bone marrow hematopoietic cells in cyclophosphamide(CTX)tumor-bearing mice,and exploring the mechanism of acupuncture and moxibustion in improving myelosuppression after chemotherapy from the perspective of numb-Notch signaling pathway.Methods:CTX was used to induce bone marrow suppression model in both healthy constitutional mice and tumor-bearing mice.Small-flux gene screening and immunohistochemistry were used to verify the significant difference genes in the numb-Notch signaling pathway of mice in each group.Effects of acupuncture and moxibustion on differentially expressed genes in bone marrow hematopoietic cells of CTX tumor-bearing mice were analyzed.Results:Genes numb1,numb2,Notch2,and jag1 which belong to the Notch signaling pathway-related genes in mice bone marrow hematopoietic cells showed significant differences.Therefore these are key genes.Acupuncture and moxibustion can up-regulate the expression of numb1 and numb2 protein and down-regulate the expression of Notch2 and jag1 protein.Conclusion:Up-regulating the expression of genes numb1 and numb2,thereby down-regulating the expression of Notch2 and jag1,which belong to the numb-Notch signaling pathway significantly differentially genes,inhibiting the over-activated Notch signaling pathway to enhance bone marrow hematopoietic function could be one of the microscopic mechanisms of acupuncture and moxitustion to reduce bone marrow suppression caused by CTX chemotherapy,and to improve hematopoietic function.