艾灸对骨髓抑制小鼠PI3K-p85、AKT、mTOR以及PTEN蛋白表达的影响

Effect of Moxibustion on Protein Expressions of PI3K-p85,AKT,mTOR and PTEN in Myelosuppressive Mice

目的:探究艾灸对穴大椎-足三里对环磷酰胺(CTX)诱导的骨髓抑制小鼠骨髓组织中PI3K-p85、AKT、mTOR以及PTEN蛋白表达的影响,从而探讨艾灸对PI3K/AKT/mTOR信号通路中关键蛋白的调控作用。方法:将105只小鼠随机分为空白组、模型组、对穴治疗组、传统配穴治疗组、rhG-CSF治疗组,每组21只。采用环磷酰胺制备小鼠骨髓抑制模型,对穴治疗组选取小鼠双侧足三里、大椎穴给予艾柱灸治疗,3壮/穴,1次/d,连续治疗7 d;传统配穴组选取小鼠双侧足三里、关元穴给予艾柱灸治疗,疗程同对穴治疗组;rhG-CSF治疗组给予重组人粒细胞集落刺激因子(rhG-CSF)治疗,1次/d,连续治疗7 d。采用Western-blot法检测小鼠骨髓组织中PI3K-p85、AKT、mTOR以及PTEN的蛋白表达。结果:PI3K-p85结果显示,模型组、对穴组和传统配穴组之间相比,差异无统计学意义(P>0.05),rhG-CSF组蛋白表达有所提升,与传统配穴组相比,差异具有统计学意义(P<0.05),相较模型组差异亦有统计学意义(P<0.01),对穴组与rhG-CSF组相比差异无统计学意义(P>0.05);AKT结果显示,除空白组外,其余各组组间无明显差异(P>0.05);mTOR结果显示,对穴组、传统配穴组、rhG-CSf组蛋白表达与模型组比较明显升高(P<0.01),rhG-CSF组与对穴组和传统配穴组相比,差异有统计学意义(P<0.01),对穴组与传统配穴组之间差异无统计学意义(P>0.05);PTEN结果显示,模型组蛋白表达升高明显(P<0.01),而对穴组、传统配穴组和rhG-CSF组组间差异无统计学意义(P>0.05)。结论:艾灸能有效调控PI3K/AKT/mTOR信号通路中PI3K-p85、AKT、mTOR、PTEN的蛋白表达,其作用效果与临床西药相近,艾灸的调控作用可能与激活PI3K/AKT/mTOR信号通路,由此调节骨髓造血干细胞的生长、代谢等,从而有效改善化疗药物造成的骨髓抑制的情况有关。

Objective:To explore the influence of moxibustion on Dazhui(DU14)and Zusanli(ST36)to the protein expressions of PI3K-p85,AKT,mTOR and PTEN in the mice with CTX-induced myelosuppression,so as to explore the regulation effect of moxibustion on key proteins of PI3K/AKT/mTOR signaling pathway.Methods:105 mice were divided into the blank group,the model group,the paired acupoints group,the traditional acupoints matching group,and the rhG-CSF treatment group,with 21 mice in each group.Myelosuppression model was established by CTX in mice.The paired acupoints group was treated with moxibustion on both sides of ST36 and DU14,once a day for 7 days;and the traditional acupoints matching group was treated with moxibustion on bilateral ST36,Guanyuan(RN4),once a day for 7 days;the rhG-CSF treatment group was given recombinant human granulocyte colony stimulating factor(rhG-CSF),once a day for 7 days.Western-blot was used to detect the protein expression of PI3K-p85,AKT,mTOR and PTEN in bone marrow tissues of the mice.Results:There was no statistical difference in PI3K-p85 among the model group,the paired acupoints group and the traditional acupoints matching group(P>0.05).The protein expression of the rhG-CSF group was increased,which was significantly different from that of the traditional acupoints matching group(P<0.05),also there was a very significant difference compared with the model group(P<0.01),and there was no statistical difference between the paired acupoints group and the rhG-CSF group(P>0.05).AKT results showed that except for the blank group,there was no significant difference among the other groups(P>0.05).mTOR results showed that the protein expression increased significantly in the paired acupoints group,the traditional acupoints matching group and the rhG-CSF group when compared with that in the model group(P<0.01),of which there was statistical difference between the rhG-CSF group and the paired acupoints group,as well as between the rhG-CSF group and the traditional acupoints matching group(P<0.01);and there was no statistical difference between the paired acupoints group and the traditional acupoints matching group(P>0.05).The results of PTEN showed that the protein expression was significantly increased in the model group(P<0.01),in which there was no statistical difference among the paired acupoints group,the traditional acupoints matching group and the rhG-CSF group(P>0.05).Conclusion:Moxibustion can effectively regulate the protein expression of PI3K-p85,AKT,mTOR,and PTEN of PI3K/AKT/mTOR signaling pathway.Its effect is similar to that of clinical western medication.The regulatory effect of moxibustion may activate PI3K/AKT/mTOR signaling pathway,thus to regulate growth and metabolism of bone marrow cells,thereby effectively improving bone marrow suppression caused by chemotherapeutic drugs.