艾灸对癌性疲劳小鼠JAK-STAT信号通路的影响

Effects of moxibustion on the expression of JAK-STAT signal pathway in cancer fatigue mice

目的观察艾灸对癌性疲劳模型小鼠及其JAK-STAT信号通路表达的影响。方法将55只BALB/C小鼠随机分为空白组10只、模型组15只、艾灸组15只及红外组15只。选择4T1乳腺癌细胞系在雌性BALB/C小鼠左腋下移植,顺铂构建癌性疲劳小鼠模型,干预4周后检测疲劳跑台实验及悬尾实验,并运用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测血清炎症细胞因子白介素-6(interleukin-1,IL-6)、肿瘤坏死因子-α(tumor necrosis factor,TNF-α)含量;实时荧光定量PCR法检测蛋白酪氨酸激酶(Janus kinase 1,Jak1)、信号传导及转录激活蛋白3(signal transducer and activator of transcription,STAT)及IL-6 mRNA表达;蛋白免疫印迹法(western blot,WB)法检测Jak1、Phospho-Jak1、STAT3、Phospho-STAT3蛋白表达。结果与空白组比较,模型组小鼠的疲劳跑台时间、距离明显减少(P<0.01),电击次数及悬尾时间显著增加(P<0.01);与模型组比较,红外组、艾灸组小鼠跑台时间、距离增加(P<0.05),跑台电击次数和悬尾不动时间减少(P<0.05);与红外组比较,艾灸组悬尾小鼠不动时间减少(P<0.05)。在观察对JAK-STAT信号通路表达情况中,与空白组比较,模型组IL-6、TNF-α含量升高(P<0.01);与模型组相比,艾灸组和红外组IL-6、TNF-α、Phospho-Jak1、Phospho-STAT3蛋白表达量及IL-6、Jak1及STAT3 mRNA表达量降低(P<0.05);与红外组相比,艾灸组IL-6、TNF-α含量,Jak1、STAT3及IL-6 mRNA表达量减少(P<0.05)。结论艾灸可能通过下调炎症因子介导的JAK-STAT信号通路,起到改善癌性疲劳的作用,而红外线是艾灸起效的因素之一。

Objective To observe the effects of moxibustion on Cancer related fatigue(CRF)model mice and the expression of JAK-STAT signal pathway.Methods 55 BALB/c mice were randomly divided into the control group,the model group,the moxibustion group and the infrared group.The 4T1 breast cancer cell line was transplanted into the female BALB/C mice under the left axillary line,and the cisplatin(5mg/kg)was used to construct the model of CRF.After 4 weeks of intervention,the mice were killed after fatigue treadmill test and tail suspension test,and method of Elisa was used to detect the levels of inflammatory cytokines IL-6 and TNF-αin the serum.The method of RT-PCR was used to detect the expression of Jak1 mRNA,STAT3 mRNA and IL-6 mRNA.The method of western blot was used to detect the protein expression of Jak1,phospho-Jak1,STAT3 and phospho-STAT3.Results Compared with the control group,the fatigue treadmill running time and distance of model group mice were significantly decreased(P<0.01),and the electric shock times of treadmill fatigue and tail suspension time were significantly increased(P<0.01).Compared with the model group,the treadmill running time and distance in infrared group and moxibustion group increased(P<0.05),and the number of electric shocks and tail suspension immobility time decreased(P<0.05).Compared with the infrared group,the immobility time of tail suspension mice in the moxibustion group was decreased(P<0.05).In observing the expression of JAK-STAT signal pathway,compared with control group,IL-6 and TNF-αwere increased in the model group(P<0.01).Compared with model group,the levels of IL-6,TNF-α,phospho-jak1,phospho-stat3,IL-6 mRNA,Jak1 mRNA and STAT3 mRNA in the moxibustion group and the infrared group were decreased(P<0.05).Compared with the infrared group,IL-6,the levels of TNF-α,Jak1 mRNA,STAT3 mRNA and IL-6 mRNA in the moxibustion group were decreased(P<0.05).Conclusion Moxibustion may relieve cancer fatigue by down regulating the expression of JAK-STAT signal pathway and inflammatory factors,and infrared ray is one of the factors.