针灸对环磷酰胺荷瘤小鼠血清细胞集落刺激因子含量的影响

Effects of acupuncture and moxibustion on serum granulocyte colony stimulating factors in CTX-induced tumor-bearing mice

目的观察针刺和艾灸前后环磷酰胺(CTX)模型荷瘤小鼠血清中粒-巨噬细胞集落刺激因子(GM-CSF)和粒细胞集落刺激因子(G-CSF)含量的变化,探讨针灸改善荷瘤小鼠骨髓抑制、升高白细胞数的作用机制。方法选用清洁级昆明种雄性小鼠64只,植种S180肉瘤造成移植瘤模型,按体重随机分为空白组、模型组、针刺组、艾灸组,每组16只。其中模型组、针刺组、艾灸组腹腔注射CTX,制备荷瘤小鼠骨髓抑制模型,空白组腹腔注射等量生理盐水。针刺组、艾灸组均选取大椎、膈俞、肾俞、足三里,每天固定时间分别进行针刺、艾灸治疗,空白组、模型组每日陪同抓取、固定,不做任何治疗。均每日1次,连续5 d。各组小鼠分别采尾部静脉血,人工镜检外周血白细胞计数。并分别于治疗3 d和治疗5 d后的第2天,每组各处死8只小鼠,摘取眼球,取其血清,运用酶联免疫吸附试验(ELISA)观察血清中GM-CSF和G-CSF的含量。结果注射CTX的各组荷瘤小鼠白细胞均降低,且第4天降至最低,血清中GM-CSF和G-CSF含量明显下降,与空白组比较有统计学意义(P<0.05)。第5天,针刺组与艾灸组白细胞开始回升,血清中的GM-CSF和G-CSF含量升高,与模型组比较有统计学意义(P<0.05)。第6天,针刺组、艾灸组白细胞数均已接近基础白细胞水平,血清GM-CSF和G-CSF含量明显升高,与模型组比较有统计学意义(P<0.05)。结论CTX造成荷瘤小鼠骨髓造血细胞损伤,导致外周血白细胞减少,且血清GM-CSF和G-CSF的含量降低。针刺和艾灸可以提高CTX荷瘤小鼠血清中GM-CSF和G-CSF的含量,诱导粒细胞前体和巨噬细胞前体细胞呈集落生长,刺激造血细胞分化,以利于髓系造血细胞的增值、分化和成熟,从而缓解化疗后的骨髓抑制,提升外周血白细胞。

Objective To study the content of serum granulocyte macrophage colony stimulating factor（GM- CSF） and granulocyte colony stimulating factor（G-CSF） in cyclophosphamide （CTX）-induced tumor-beating mice before and after acupuncture and moxibustion treatment, so as to find out the mechanism of acupuncture and moxibustion treatment on recovery from myelosuppression and increasing of leukocyte counts. Methods 64 Kunming male mice of clean grade were transplanted subcutaneously S180 sarcoma cells to induce the trans- planting tumor model, then were randomly divided into control group, model group, acupuncture group and moxibustion group （each n = 16） according to the body weight. Mice in model group, acupuncture group and moxibustion group were intraperitoneally injected with CTX to establish CTX-induced tmnor-bearing mice mod- el, and the control group were injected intraperitoneally with normal saline. Acupoints Dazhui（GV14）, Geshu （ BL17 ）, Shenshu （ BL23 ） and Zusanli （ST36） were needled in acupuncture group and were moxa moxibusted in moxibustion group once daily at regular time for consecutive 5 d. Peripheral leukocyte counts were meas- ured by detecting the blood collected from caudal vein before modeling and at every day of treatment proce- dure. The serum concentrations of GM-CSF and G-CSF were determined by using ELISA method at the third day of treatment and one day after five-day-treatment. Results Compared with control group, peripheral leu- kocyte counts decreased, even to the lowest level at the fourth day in the three groups of CTX-induced tumor- bearing mice, and the serum concentrations of GM-CSF and G-CSF also decreased in the three groups signifi- cantly（P 〈0.05）. At the fifth day of treatment, leukocyte counts started to increase, and the serum concen- trations of GM-CSF and G-CSF increased significantly compared with model group（P 〈0.05）. At the day after treatment, leukocyte counts increased to nearly normal level and the serum concentrations of GM-CSF and G- CSF increased dramatically compared with model group. Conclusion CTX damaged the myeloid element to reduce peripheral leukocyte counts and the serum concentrations of GM-CSF and G-CSF. Acupuncture and moxibustion can obviously increase the concentration of GM-CSF and G-CSF, that is to say, both the Chinese medical treatment can induce colony growth of the granulocyte precursor and macrophage precursor cells and stimulate differentiation of the hematopoietic cells, leading to proliferation, differentiation and maturation of the myeloid hematopoietic cells, resulting in recovery from myelosuppression and increasing of leukocyte counts induced by chemotherapy.