艾灸调节肉瘤微环境中Th1/Th2漂移的作用

Effect of Moxibustion Regulating Th1/Th2 Drift in TME of Sarcoma

目的:观察艾灸对肉瘤微环境中Th1细胞、Th2细胞和Th1/Th2漂移的影响。方法:SPF级C57BL/6J小鼠随机分为空白组、模型组、顺铂组和艾灸组,每组10只。采用S180肉瘤细胞0.1 mL(1×10^(7)/mL)接种于C57BL/6J小鼠胸部皮下形成移植瘤。以移植瘤为穴位中心,采用热敏艾条治疗,距离3 cm,每次艾灸10 min,每日1次,连续治疗14 d为一疗程。顺铂组给以顺铂腹腔注射治疗。治疗一疗程后小鼠处死,Luminex液相悬浮芯片检测血清IL-2、IFN-γ、IL-4和IL-10的浓度。检测移植瘤的重量,流式细胞技术检测小鼠脾Th1和Th2细胞的数量,荧光原位杂交技术(Fish)检测移植瘤微环境中Th1和Th2的基因表达。结果:与模型组比较,艾灸组抑制肉瘤的生长,差异具有统计学意义(P<0.01),和顺铂组比较,两组差异无统计学意义(P>0.05)。与模型组和顺铂组比较,艾灸组提高小鼠血清IL-2和IFN-γ的浓度,差异具有统计学意义(P<0.05),降低IL-10和IL-4的浓度,差异具有统计学意义(P<0.01)。与模型组和顺铂组比较,艾灸增加小鼠脾Th1细胞的数量,差异具有统计学意义(P<0.01),降低Th2细胞的数量,差异具有统计学意义(P<0.05)。与模型组和顺铂组比较,艾灸组上调移植瘤微环境中Th1基因的表达,差异具有统计学意义(P<0.01),抑制Th2基因的表达,差异具有统计学意义(P<0.01)。结论:艾灸可抑制S180肉瘤的生长;提高血清IL-2和IFN-γ的浓度,降低IL-10和IL-4的浓度;增加小鼠脾Th1细胞的数量,降低Th2细胞的数量;提高移植瘤微环境中的Th1/Th2的比值,使其由Th2向Th1漂移,起到抗肿瘤的作用。

Objective:To observe the effect of moxibustion on Th1 cells,Th2 cells and Th1/Th2 drift in the tumor microenvironment(TME)of sarcoma.Methods:SPF C57BL/6J mice were randomly divided into the blank group,the model group,the Cisplatin group and the moxibustion group,with 10 mice in each group.S180 sarcoma cells were used at a concentration of 1×10^(7)/ml,0.1 ml was inoculated into C57BL/6J mice subcutaneously to form xenograft tumors.Focusing on the xenograft tumor,the heat-sensitive moxa stick was used to treat the xenograft tumor at a distance of 3 cm,10 minutes each time,once a day for 14 consecutive days.The Cisplatin group was given intraperitoneal injection of Cisplatin.After 14 days of the treatment,the mice were sacrificed,and the serum levels of IL-2,IFN-γ,IL-4 and IL-10 were detected by Luminex liquid suspension chip.The weight of the xenograft tumor was measured,the numbers of Th1 cells and Th2 cells of spleen were measured by flow cytometry,and the gene expressions of Th1 and Th2 in the microenvironment of the xenograft tumor were detected by fluorescence in situ hybridization(Fish).Results:Compared to that in the model group,the growth of sarcoma was significantly inhibited in the moxibustion group(P<0.01);however,there was no statistical differences in the growth of sarcoma between the moxibustion group and the Cisplatin group(P>0.05).The serum concentrations of IL-2 and IFN-γwere obviously increased(P<0.05),and the concentrations of IL-10 and IL-4 were obviously decreased in the moxibustion group compared to those in the model group and the Cisplatin group(P<0.01).Compared to those in the model group and the Cisplatin group,the number of Th1 cells was greatly increased(P<0.01),however the number of Th2 cells was greatly decreased in the moxibustion group(P<0.05).Compared to those in the model group and the Cisplatin group,the gene expression of Th1 in the microenvironment of xenograft tumors was significantly up-regulated(P<0.01),but the gene expression of Th2 was significantly inhibited in the moxibustion group(P<0.01).Conclusion:Moxibustion can inhibit the growth of S180 sarcoma,increase the serum concentrations of IL-2 and IFN-γ,decrease the concentrations of IL-10 and IL-4,increase the number of Th1 cells and decrease the number of Th2 cells in mice spleen,as well as improve the ratio of Th1/Th2 in the microenvironment of xenograft tumors,making it drift from Th2 to Th1 to achieve the effect of anti-tumor.