针刺对免疫抑制的Cx43基因敲除小鼠免疫功能的影响

目的:观察针刺对免疫抑制的Cx43基因敲除小鼠(Cx43+/-鼠)免疫功能的影响,探讨缝隙连接蛋白43(Cx43)与针刺信号传递机制之间的可能关系。方法:8~9周龄雄性Cx43+/-小鼠及野生型小鼠(Cx43+/+鼠)各18只,分别随机分为正常组、模型组和针刺组,每组6只。小鼠腹腔注射环磷酰胺(80 mg/kg)制造免疫抑制模型。针刺双侧"足三里"穴及"关元"穴,每5 min行针1次,每次30 s,留针15 min,连续治疗7 d。计算各组小鼠的脾脏指数及胸腺指数和外周血白细胞数,并采用流式细胞术检测脾脏T淋巴细胞亚群的百分数。结果:Cx43+/+鼠及Cx43+/-鼠的模型组与各自的正常对照组相比,其脾脏指数、胸腺指数、白细胞数及CD3+%、CD4+%均明显降低(P<0.05,0.01),提示两种动物模型组免疫功能显著低下。在Cx43+/+小鼠上,针刺组的脾脏指数、胸腺指数、白细胞数、CD3+%、CD4+%、CD8+%与模型组比较均明显升高(P<0.05,0.01),与正常组比较差异无显著性意义(P>0.05),提示针刺可增强免疫功能;而对于Cx43+/-小鼠,针刺组脾脏指数、胸腺指数、白细胞数、CD3+%、CD4+%、CD8+%与模型组相比较差异无显著性意义(P>0.05),说明基因Cx43敲除后针刺的效应消失。结论:Cx43基因的敲除抑制了针刺改善免疫力效应的产生,提示以Cx43为主要组成的细胞间缝隙连接(GJ)可能在针刺调整机体免疫功能的信号传递通路上起着重要的作用,GJ可能与针刺改善免疫力效应的产生有着密切的关系。

Influence of Acupuncture on the Immune Function of Immunosuppressive Cx43 Knockout Mice

Objective: To observe the influence of acupuncture on the immune function in immunosuppressive Cx43 knockout mice so as to explore the relationship between connexin 43(Cx43) and acupuncture signal transmission.Methods: Eighteen male Cx43 knockout mice(Cx43 /-,8-9 weeks old) and eighteen wide-type mice(Cx43 / ) were divided into normal group,model group and acupuncture group randomly,with 6 cases in each.The immunosuppressive mice model was established by intraperitoneal injection of Cyclophosphamide.Mice in acupuncture group were treated with manual acupuncture of "Zusanli"(ST 36) and "Guanyuan"(CV 4),once daily for 7 days.Spleen index,thymus index,number of leukocytes in the peripheral blood were calculated and the percentage of T lymphocyte subpopulation was detected by using flow cytometry.Results: Compared with normal control group,the spleen index,thymus index and leukocyte number of model group in both Cx43 / mice and Cx43 /-mice were obviously lower(P<0.05,0.01),while in Cx43 / mice and in comparison with model group,the spleen index,thymus index and leukocyte number of acupuncture group were significantly higher(P<0.05,0.01) and no significant differences were found between acupuncture group and control group(P>0.05).But in Cx43 /-mice,no obvious differences were found between acupuncture and model groups in the aforementioned 3 indexes(P>0.05),showing a disappearance of the effect of acupuncture in raising immune function after Cx43 knockout.There were no obvious differences between two control groups and between two model groups in the 3 indexes of Cx43 / mice and Cx43 /-mice.The immune function of Cx43 / mice was obviously stronger than that of Cx43 /-mice in acupuncture group.Conclusion: Knockout of Cx43 gene inhibits the effect of acupuncture in reinforcing mouse immunity,suggesting an important role of Cx43 in the transmission of acupuncture signals for adjusting immunity function.