针灸对环磷酰胺化疗小鼠血清中FL与MIP-1α含量影响的研究

发布时间：2018-07-04 23:27

本文选题：环磷酰胺 + 骨髓抑制　；参考：《河南中医药大学》2016年硕士论文

【摘要】：目的通过观察在针刺和艾灸不同作用下,环磷酰胺(Cyclophosphamide,CTX)化疗小鼠血清中造血细胞因子FIt3配体(flt3 ligand,FL)与巨噬细胞炎性蛋白-1α(macrophage inflammatory protein-1α,MIP-1α)的变化,阐释针灸改善CTX化疗小鼠骨髓造血微环境,保护造血干细胞,提升外周血白细胞的分子生物学机制,为针灸改善化疗所致小鼠造血微环境损伤方面提供实验研究依据。方法选取SPF级雄性昆明种小鼠80只(7周龄、体重22±2g),根据治疗天数不同依据体重分层随机分两大组,即3d组和5d组,每组40只。再按其体重分层随机分成8小组:空白组(A3组、A5组)、模型组(B3组、B5组)、针刺组(C3组、C5组)、艾灸组(D3组、D5组),每亚组各10只。空白组根据小鼠体重按照浓度0.02ml/g给予生理盐水腹腔注射,连续3d。其余4组腹腔注射CTX,根据小鼠体重0.02ml/g,注射CTX浓度为5mg/ml,即100mg/kg/d,连续3d,停药后4小时骨髓抑制模型即成。治疗时空白组、模型组陪同固定,不做治疗。针刺组、艾灸组选取“大椎”、“膈俞”、“肾俞”、“足三里”四穴给予针刺和艾灸治疗,连续治疗3d和5d。实验期间每日定时专人割破鼠尾静脉取血,镜检外周血白细胞计数。各组小鼠分别在治疗3d、5d后的第2d摘眼球取血,离心,取上清液,取材后放入-80℃低温冰箱保存、备检。送检材料采用ELISA法检测小鼠血清中FL和MIP-1α的含量。运用SPSS20.0 for Windows软件进行数据统计分析。结果1造模后,与空白组对照,剩余3组外周血白细胞数量大幅度降低,有统计学差异(P0.05),且3组之间降低幅度相差不多,说明骨髓抑制模型成功。治疗3d后,与模型组比较,针刺组和艾灸组白细胞计数有升高趋势,但无统计学差异(P0.05)。治疗5d后,针刺组和艾灸组白细胞计数已达到并超过空白组,有统计学差异(P0.05),与模型组比较,有统计学差异(P0.05)。2经过3d的治疗,与模型组比较,针刺组和艾灸组中血清中FL含量表达均降低,有统计学差异(P0.05);与针刺组比较,艾灸组中的含量表达偏低,但无统计学差异(P0.05)。治疗5天后,针刺组与艾灸组中含量表达仍低于模型组,有统计学差异(P0.05);与针刺组比较,艾灸组含量表达依然偏低,仍无统计学差异(P0.05)。3经过3d治疗,与模型组相比,针刺组和艾灸组血清中MIP-1α含量表达均高于模型组,有统计学差异(P0.05);与针刺组对比,艾灸组表达含量偏高,但无统计学差异(P0.05)。治疗5d后,与模型组比较,针刺组和艾灸组中的含量表达均高于模型组,有统计学差异(P0.05);与针刺组比较,艾灸组中含量表达仍偏高,但无统计学差异(P0.05)。结论1针灸改善化疗后骨髓抑制,升高白细胞的疗效是确切的,再次佐证导师组前期研究成果及规律,针刺和艾灸可使CTX化疗小鼠白细胞较模型组提前1d回升并提前1d超过基础水平。2化疗后小鼠血清中FL的含量表达明显升高,针刺、艾灸可以下调化疗小鼠血清中FL的含量表达,并且将FL水平维持在适当浓度,以利于早期造血细胞的存活、增殖、分化,修复损伤的造血微环境,改善化疗后骨髓抑制。3化疗后小鼠血清中MIP-1α的表达含量降低。通过针刺、艾灸的作用,化疗小鼠血清中MIP-1α的含量表达升高,既可以选择性的保护造血干细胞,又能协同集落刺激因子促集落的形成,以促进骨髓造血干/祖细胞的增殖和成熟,改善骨髓造血微环境,提升外周血白细胞计数。4针灸可促进化疗后骨髓抑制尽快得到改善,损伤的造血微环境能够得到恢复,且治疗作用可随治疗天数的增加而增加,具有治疗意义,艾灸作用优于针刺。
[Abstract]:Objective To observe the changes of hematopoietic cell factor FIt3 ligand (Flt3 ligand, FL) and macrophage inflammatory protein -1 alpha (macrophage inflammatory protein-1 a, MIP-1 alpha) in the serum of mice treated with cyclophosphamide (Cyclophosphamide, CTX) under different effects of acupuncture and moxibustion, and to explain the improvement of hematopoietic microenvironment and protection of bone marrow in mice with chemotherapy by Acupuncture and moxibustion. Hematopoietic stem cells, promoting the molecular biological mechanism of peripheral blood leucocyte to provide experimental basis for the improvement of hematopoietic microenvironment injury induced by chemotherapy in mice. Methods 80 male Kunming mice (7 weeks of age, 22 + 2G) were selected and divided into two groups according to the number of days of treatment, that is, group 3D and 5D, each group. Group 40 were divided randomly into 8 groups according to their weight stratification: blank group (group A3, group A5), model group (group B3, B5), acupuncture group (group C3, C5), moxibustion group (group D3, D5), 10 in each subgroup. The blank group was given abdominal injection of normal saline on the weight of mice according to the concentration 0.02ml/g, and the rest of the other 4 groups were injected CTX, according to mice weight 0.02ml/g. The concentration of CTX was 5mg/ml, 100mg/kg/d, continuous 3D, and the bone marrow suppression model was formed 4 hours after the drug stopped. The time and space white group was treated. The model group was accompanied by fixed, no treatment. The acupuncture group, the moxibustion group selected the four points of "Da Zhui", "Shenshu", "Shenshu", "Zusanli", given acupuncture and moxibustion for the continuous treatment of 3D and 5d. experiments. During the treatment of 3D and 5D, the mice were removed from the rat's tail vein to take blood and take the blood from the 3D and 5D after the treatment. After the centrifugation, the supernatant was centrifuged, and then stored at -80 centigrade cryogenic refrigerator. The content of FL and MIP-1 alpha in the serum of mice was detected by ELISA. SPSS20.0 for Windows software was used. Data statistical analysis. Results after 1 models, compared with the blank group, the number of white blood cells in the remaining 3 groups decreased significantly (P0.05), and the difference between the 3 groups was not much, indicating that the bone marrow suppression model was successful. After the treatment of 3D, the white blood cell count of the acupuncture group and the moxibustion group was higher than that in the model group, but there was no statistical difference. Difference (P0.05). After the treatment of 5D, the white blood cell count of the acupuncture group and the moxibustion group had reached and exceeded the blank group. There was a statistical difference (P0.05). Compared with the model group, there was a statistical difference (P0.05).2 after 3D treatment. Compared with the model group, the expression of FL content in the serum of acupuncture group and moxibustion group decreased, with statistical difference (P0.05), compared with the acupuncture group, and compared with the acupuncture group, and compared with the acupuncture group. Compared with the moxibustion group, the expression of the content was low, but there was no statistical difference (P0.05). The contents of the acupuncture group and the moxibustion group were still lower than the model group (P0.05) after 5 days of treatment. Compared with the acupuncture group, the content of the moxibustion group was still low, and there was still no difference (P0.05).3 after 3D treatment. Compared with the model group, the acupuncture group and moxibustion were compared with the model group. The expression of MIP-1 alpha in the serum of the group was higher than that in the model group (P0.05). Compared with the acupuncture group, the expression level of the moxibustion group was high, but there was no statistical difference (P0.05). After the treatment of 5D, the content expression in the acupuncture group and moxibustion group was higher than that in the model group, with a statistical difference (P0.05); compared with the acupuncture group, the moxibustion group was compared with the moxibustion group. The expression of the content was still high, but there was no statistical difference (P0.05). Conclusion 1 the effect of acupuncture and moxibustion on the improvement of bone marrow suppression after chemotherapy and the increase of white blood cells is exact. Again, the results and rules of the early study of the tutor group are confirmed again. Acupuncture and moxibustion can make the leukocytes in the CTX chemotherapy mice advance 1D ahead of time in the model group and advance to the blood of the mice after the basic level of.2 chemotherapy. The content expression of FL in the Qing Dynasty increased obviously. Acupuncture and moxibustion could reduce the expression of FL in the serum of chemotherapy mice, and maintain the level of FL at the appropriate concentration to help the survival, proliferation, differentiation and repair of the hematopoietic microenvironment of the early hematopoietic cells, and improve the expression of MIP-1 alpha in the serum of mice after chemotherapy after chemotherapy, and to improve the expression of MIP-1 alpha in the serum of mice after.3 chemotherapy. Under the effect of acupuncture and moxibustion, the expression of MIP-1 alpha in serum of chemotherapy mice increased, which could selectively protect hematopoietic stem cells and synergy the formation of colony stimulating factors to promote the proliferation and maturation of bone marrow hematopoietic stem / progenitor cells, improve the microenvironment of bone marrow blood making and increase the peripheral blood white blood cell count.4 acupuncture and moxibustion After the promotion of chemotherapy, bone marrow suppression is improved as soon as possible. The hematopoietic microenvironment can be recovered, and the therapeutic effect can be increased with the increase of the number of days of treatment. It is of therapeutic significance. Moxibustion is superior to acupuncture.
【学位授予单位】：河南中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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