针灸对荷瘤小鼠脾淋巴细胞增殖及Stat5-Foxp3通路影响

Effect of Acupuncture and Moxibustion on Splenic Lymphopoiesis and Stat5-Foxp3 Pathway in Tumor Mice

目的探索针灸"大椎穴"调节肿瘤免疫抑制的作用机制。方法将48只雄性Balb/c小鼠随机分为电针组、艾灸组、荷瘤组和正常组,每组12只。在小鼠左腋皮下接种H22肿瘤细胞,建立肿瘤模型。分别采用电针、艾灸大椎治疗。观察生存率、抑瘤率、脾脏指数、胸腺指数。AlamarBlue法检测ConA刺激的小鼠脾淋巴细胞增殖,增殖能力以还原值表示。RT-PCR方法检测脾淋巴细胞Foxp3 mRNA、Stat5a mRNA、Stat5b mRNA以及胸腺Foxp3 mRNA表达。结果艾灸组荷瘤小鼠生存率高于荷瘤组,但差异无统计学意义(P>0.05);艾灸组抑瘤率为39.3%;电针组与艾灸组荷瘤小鼠脾脏指数较荷瘤组升高(P<0.05);电针组与艾灸组荷瘤小鼠胸腺指数较荷瘤组胸腺指数升高,但差异无统计学意义(P>0.05)。AlamarBlue法检测电针组、艾灸组、荷瘤组小鼠脾淋巴细胞增殖还原值均高于正常组(P<0.05);艾灸组还原值高于荷瘤组(P<0.05);荷瘤组小鼠脾淋巴细胞Foxp3 mRNA、Stat5a mRNA、Stat5b mRNA表达较正常组增高(P<0.05);艾灸组、电针组小鼠脾淋巴细胞Foxp3 mRNA、Stat5a mRNA、Stat5b mRNA较荷瘤组表达降低(P<0.05);荷瘤组小鼠胸腺Foxp3 mRNA表达较正常组增高(P<0.05);艾灸组小鼠胸腺Foxp3 mRNA表达较荷瘤组降低(P<0.05);电针组小鼠胸腺Foxp3 mRNA表达较荷瘤组降低,但差异无统计学意义(P>0.05)。结论艾灸治疗具有抑制H22荷瘤小鼠瘤体生长、提高H22荷瘤小鼠生存率的趋势;电针和艾灸治疗能提高H22荷瘤小鼠脾脏指数、增强H22荷瘤小鼠脾淋巴细胞增殖能力;针灸调节肿瘤免疫抑制效应与电针和艾灸下调H22荷瘤小鼠脾淋巴细胞Foxp3 mRNA、Stat5a mRNA、Stat5b mRNA表达,胸腺Foxp3 mRNA表达有关。

Objective To explore the mechanism of regulative action of acupuncture and moxibustion at point Dazhu on tumor immunosuppression. Method Forty-eight male Balb/c mice were randomly allocated to electroacupuncture, moxibustion, tumor and normal groups, 12 mice each. A mouse model of tumor was made by its left axillary subcutaneous inoculation of H22 tumor cells. Electroacupuncture and moxibustion at point Dazhu were given respectively. The survival rate, tumor suppression rate, spleen index and thymus index were examined. ConA-stimulated mouse splenic lymphopoiesis was examined by Alamar Blue assay. The proliferative capacity was represented by a reduced value. The expressions of Foxp3 mRNA, Stat5a mRNA and Stat5b mRNA in splenic lymphocytes and Foxp3 mRNA in the thymus gland were examined by the RT-PCR method. Results The survival rate was higher in the moxibustion group of tumor mice than in the tumor group but there was no statistically significant difference （P〉0.05）. The tumor suppression rate was 39.3%. The spleen index was higher in the electroacupuncture and moxibustion groups of tumor mice than in the tumor group （P〈0.05）. The thymus index was higher in the electroacupuncture and moxibustion groups of tumor mice than in the tumor group but there was no statistically significant difference （P〉0.05）. The Alamar Blue assay showed that the reduced value of mouse splenic lymphopoiesis was higher in the electroacupuncture, moxibustion and tumor groups of mice than in the normal group （P〈0.05）. The reduced value was higher in the moxibustion group than in the tumor group （P〈0.05）. The expressions of Foxp3 mRNA, Stat5a mRNA and Stat5b mRNA in splenic lymphocytes increased in the tumor group of mice compared with the normal group （P〈0.05） and decreased in the moxibustion and electroacupuncture groups of mice compared with the tumor group （P〈0.05）. The expression of Foxp3 mRNA in the thymus gland increased in the tumor group of mice compared with the normal group （P〈0.05） and decreased in the moxibustion group of mice compared with the tumor group （P〈0.05）. The expression of Foxp3 mRNA in the thymus gland decreased in the electroacupuncture group of mice compared with the tumor group, but there was no statistically significant difference （P〉0.05）. Condusian Moxibustion treatment tends to inhibit tumor growth and raise the survival rate in H22 tumor mice. Electroacupuncture and moxibustion treatments can increase the spleen index and improve the proliferative capacity of splenic lymphocytes in H22 tumor mice. The regulative effects of electroacupuncture and moxibustion on tumor immunosuppression are related to their downregulation of the expressions of Foxp3 mRNA, Stat5a mRNA and Stat5b mRNA in the splenic lymphocytes and Foxp3 mRNA in the thymus gland of H22 tumor mice.