不同穴位针刺对免疫抑制大鼠神经肽相关基因表达及免疫调节作用的分子机制研究
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摘要
目的观察不同穴位针刺对免疫抑制大鼠神经肽相关基因表达影响，探讨其免疫调节机制，阐明针刺对免疫抑制机体神经内分泌免疫网络的作用。
方法采用环磷酰胺腹腔注射法制备免疫抑制大鼠模型，通过RT-PCR法观察不同穴位针刺后免疫抑制大鼠脑组织β-EPmRNA、SPmRNA表达的变化；采用We s t e r n - b l o t法观察脾脏淋巴细胞表面β-EPR及SPR表达情况；通过流式细胞仪检测血中CD4+T细胞和CD8+T细胞的含量；采用ELISA法检测针刺对各组大鼠血清IL-4、IFN-γ含量的影响。
结果不同穴位针刺能够提高免疫抑制大鼠脑垂体β-EPmRNA、下丘脑SPmRNA表达，治疗各组与模型对照组比较有显著性差异（P <0 . 0 5），头针加足三里、关元组( E组)效果最佳。针刺能够降低外周脾淋巴细胞表面SPR表达，提高外周脾淋巴细胞表面β-EPR表达；同时不同穴位针刺能提高外周血CD4+T细胞含量，治疗各组与模型对照组比较有显著性差异（P <0.05），头针加足三里、关元组效果最佳，与正常组比较无显著性差异（P > 0 . 0 5）、对CD8+T细胞无显著影响，与正常组比较无显著性差异（P> 0 . 0 5）；不同穴位针刺能显著降低免疫抑制大鼠血清IL-4水平，治疗各组与模型对照组比较有显著性差异（P <0.05），头针加足三里、关元组效果最佳，还可提高血清I FN-γ含量,治疗各组与模型对照组比较有显著性差异（P <0 . 0 5），头针加足三里、关元组效果最佳。
结论不同穴位针刺可以提高免疫抑制大鼠低下的细胞免疫功能，尤其以头穴加体穴效果最佳。其可能的机理是通过影响神经肽-免疫细胞受体-细胞因子通路完成的。
Objective To observe the effect of different acupuncture points in immunosuppressive rats on the neuropeptide gene expression,e-xplore the immu ne regulatory me chanisms,to clarify role of ac-upuncture on the neuroendocrine-immune network.
Methods Preparation immunosuppressive rats model by intrape-ritoneal injection of CP .To observedβ-EPmRNA, SPmRNA exp-ression changes in brain tissue of rats by RT-PCR after acupun-cture;observed spleen cell surfaceβ-EPR and SPR expression us-ing western-blot; by FCM observed CD4+T cells and CD8+ T c-ells in blood; using ELISA method to observed IL-4, IFN-γlev-els of serum.
Results Different points acupuncture can improveβ-EPmRNA、SPmRNA expression in immunosuppressive rats, compared to mod-el group,there is a significant difference (P<0.05), E group is t-he best. Acupuncture can reduce SPR expression on the splenic lymphocytes surface,improveβ-EPR content; different points acu-puncture can improve peripheral CD4+T cells content , compare-d to model group, there is a significant difference (P<0.05).CD8+T cells had no significant difference. Different points acupuncture can significantly decrease serum IL-4, increase serum IFN-γcontent of immunosuppressive rats, compared to model group,there is a significant difference (P<0.05),E group is the best.
Conclusions Acupuncture can improve the cellular-immune funct-oin of i mmunos uppressi ve rats, particular ly the head plus body acupuncture points(E group) is best. The possible mechanis m is that affects neuropeptide-immune cell receptors-cytokins pathway.
方法采用环磷酰胺腹腔注射法制备免疫抑制大鼠模型，通过RT-PCR法观察不同穴位针刺后免疫抑制大鼠脑组织β-EPmRNA、SPmRNA表达的变化；采用We s t e r n - b l o t法观察脾脏淋巴细胞表面β-EPR及SPR表达情况；通过流式细胞仪检测血中CD4+T细胞和CD8+T细胞的含量；采用ELISA法检测针刺对各组大鼠血清IL-4、IFN-γ含量的影响。
结果不同穴位针刺能够提高免疫抑制大鼠脑垂体β-EPmRNA、下丘脑SPmRNA表达，治疗各组与模型对照组比较有显著性差异（P <0 . 0 5），头针加足三里、关元组( E组)效果最佳。针刺能够降低外周脾淋巴细胞表面SPR表达，提高外周脾淋巴细胞表面β-EPR表达；同时不同穴位针刺能提高外周血CD4+T细胞含量，治疗各组与模型对照组比较有显著性差异（P <0.05），头针加足三里、关元组效果最佳，与正常组比较无显著性差异（P > 0 . 0 5）、对CD8+T细胞无显著影响，与正常组比较无显著性差异（P> 0 . 0 5）；不同穴位针刺能显著降低免疫抑制大鼠血清IL-4水平，治疗各组与模型对照组比较有显著性差异（P <0.05），头针加足三里、关元组效果最佳，还可提高血清I FN-γ含量,治疗各组与模型对照组比较有显著性差异（P <0 . 0 5），头针加足三里、关元组效果最佳。
结论不同穴位针刺可以提高免疫抑制大鼠低下的细胞免疫功能，尤其以头穴加体穴效果最佳。其可能的机理是通过影响神经肽-免疫细胞受体-细胞因子通路完成的。
Objective To observe the effect of different acupuncture points in immunosuppressive rats on the neuropeptide gene expression,e-xplore the immu ne regulatory me chanisms,to clarify role of ac-upuncture on the neuroendocrine-immune network.
Methods Preparation immunosuppressive rats model by intrape-ritoneal injection of CP .To observedβ-EPmRNA, SPmRNA exp-ression changes in brain tissue of rats by RT-PCR after acupun-cture;observed spleen cell surfaceβ-EPR and SPR expression us-ing western-blot; by FCM observed CD4+T cells and CD8+ T c-ells in blood; using ELISA method to observed IL-4, IFN-γlev-els of serum.
Results Different points acupuncture can improveβ-EPmRNA、SPmRNA expression in immunosuppressive rats, compared to mod-el group,there is a significant difference (P<0.05), E group is t-he best. Acupuncture can reduce SPR expression on the splenic lymphocytes surface,improveβ-EPR content; different points acu-puncture can improve peripheral CD4+T cells content , compare-d to model group, there is a significant difference (P<0.05).CD8+T cells had no significant difference. Different points acupuncture can significantly decrease serum IL-4, increase serum IFN-γcontent of immunosuppressive rats, compared to model group,there is a significant difference (P<0.05),E group is the best.
Conclusions Acupuncture can improve the cellular-immune funct-oin of i mmunos uppressi ve rats, particular ly the head plus body acupuncture points(E group) is best. The possible mechanis m is that affects neuropeptide-immune cell receptors-cytokins pathway.
引文
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