针灸对CTX化疗小鼠骨髓造血微环境中骨髓基质细胞细胞周期的影响

Effect of Acupuncture and Moxibustion on Cell Cycle of BMSC in Bone Marrow Hematopoietic Microenvironment of Mice Received CTX Chemotherapy

目的:探讨针灸抗化疗毒副反应,改善骨髓造血微环境中骨髓基质细胞支持造血功能的分子生物学机制。方法:40只SPF级雄性昆明种小鼠,随机分为空白组、模型组、针刺组、艾灸组。采用国际公认的环磷酰胺(CTX)骨髓抑制模型。针刺组、艾灸组分别采用针刺、艾灸治疗,每天1次,连续5 d。空白组、模型组每天陪同针刺组、艾灸组抓取、固定,不做任何处理。每组分别于治疗后第1天、第3天、第5天随机处死3只小鼠,采用石蜡包埋小鼠左侧股骨,应用石蜡包埋组织单细胞悬液制备方法,制备成小鼠骨髓基质细胞单细胞悬液,经流式细胞仪检测小鼠骨髓造血微环境中骨髓基质细胞的细胞G1期、S期、G2期细胞百分率。结果:与模型组比较,针刺组和艾灸组可以降低CTX化疗小鼠骨髓造血微环境中骨髓基质细胞(BMSC)G1期细胞含量,其中3 d组、5 d组差异均有统计学意义(P<0.05)。与模型组比较,针刺和艾灸可以提高3 d组BMSC的S期细胞含量,针刺和艾灸可以降低5 d组BMSC的S期细胞含量,差异均具有统计学意义(P<0.05)。与模型组比较,针刺组和艾灸组可以提高1 d组、3 d组、5 d组BMSC的G2期细胞含量,差异均具有统计学意义(P<0.05)。结论:烷化剂CTX损伤骨髓造血微环境的同时,破坏了骨髓基质细胞的细胞周期规律,从而产生骨髓抑制,严重影响骨髓造血功能。针刺与艾灸可以促进CTX化疗后骨髓基质细胞(BMSC)的细胞修复,加速骨髓基质细胞从G1期向S期的转化,增强骨髓基质细胞DNA的合成,延长G2期,同时利用细胞G2期阻滞的时机,尽快修复受损DNA,加速细胞有丝分裂,进入增殖状态,提高骨髓基质细胞的修复能力,保护骨髓造血微环境,促进骨髓造血功能的重建。

Objective:To explore the effect of acupuncture and moxibustion in treating adverse reactions of chemotherapy,and to study its molecular mechanisms of improving the bone marrow stromal cells(BSMC)to support hematopoietic function.Methods:40 SPF male mice were randomly divided into the control group,the model group,the acupuncture group and the moxibustion group.The bone marrow suppression models were established by cyclophosphamide(CTX).Three rats were randomly sacrificed after one day of treatment,after three days of treatment and after five days of treatment respectively.Single cell suspension of BSMC was prepared to detect the percentage of BSMC during G1 phase,S phase and G2 phase by flow cytometry.Results:The content of BMSC of G1 phase was decreased in the acupuncture group and in the moxibustion group compared to that in the model group;of which there was statistical difference between after three days of treatment and after five days of treatment(P<0.05).The content of BMSC of S phase was increased after three days of treatment and it was decreased after five days of treatment in the acupuncture group and in the moxibustion group(P<0.05).The content of BMSC of G2 phase was increased in the acupuncture group and in the moxibustion group compared to that in the model group(P<0.05).Conclusion:Alkylating agent CTX can damage bone marrow microenvironment,and it can undermine the cycle of bone marrow stromal cells to produce bone marrow suppression at the same time,which seriously affects the bone marrow hematopoietic function.Acupuncture and moxibustion can promote BMSC repair after CTX chemotherapy,the therapies can accelerate the transformation of BMSC from G1 phase to S phase,increase of the DNA synthesis of BMSC and extend G2 phase;damaged DNA can be repaired as soon as possible during G2 arrest time.The therapies can accelerate cell mitosis into proliferation,enhance the repair capacity of BMSC,protect bone marrow microenvironment and promote reconstruction of bone marrow function.