天灸“肺俞”穴对健康大鼠Th17/Treg免疫平衡及炎症反应的影响

Effects of Natural Moxibustion at"Feishu(BL 13)"on Th17/Treg Immune Balance and Inflammatory Response in Healthy Rats

目的探讨天灸“肺俞”穴对健康大鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响。方法将48只大鼠随机分为假天灸组10只,天灸组38只。假天灸组大鼠将空白的穴位贴贴敷于双侧“肺俞”穴,天灸组大鼠将填有复方斑蝥膏的穴位贴贴敷于双侧“肺俞”穴,贴敷时间为8 h。8 h后天灸组大鼠成功发泡30只,再随机分为天灸1天组、天灸3天组、天灸7天组,每组10只。实验期间记录大鼠一般情况;天灸不同时间组在相应时间取材,采用HE染色观察大鼠贴敷区域皮肤的病理改变;采用流式细胞术检测外周血Th17、Treg细胞表达,计算Th17/Treg值;采用ELISA法检测血清白细胞介素17A(IL-17A)、白细胞介素6(IL-6)、白细胞介素10(IL-10)含量。结果与假天灸组比较,天灸1天组大鼠贴敷区域可见水泡,大鼠经常搔抓天灸区域皮肤,皮肤出现角质层松散,棘层、颗粒层增厚,基底层细胞缺失,炎性细胞浸润,外周血Treg及血清IL-17A、IL-6水平升高(P<0.01);天灸3天组天灸区域结痂并可见部分脱落,皮肤病理改变最明显,外周血Th17、Treg表达及血清IL-17A、IL-6、IL-10水平升高(P<0.01);天灸7天组天灸区域皮肤损伤及病理改变基本恢复。天灸1、3、7天组Th17/Treg值均较假天灸组降低(P<0.05或P<0.01)。与天灸1天组比较,天灸3天组外周血Th17、Treg表达及血清IL-17A升高;天灸7天组外周血Treg表达及血清IL-17A、IL-6降低,Th17/Treg值升高(P<0.05或P<0.01)。与天灸3天组比较,天灸7天组外周血Th17、Treg表达及血清IL-17A、IL-6、IL-10降低,Th17/Treg值升高(P<0.05或P<0.01)。结论天灸“肺俞”穴能使健康大鼠Th17/Treg平衡向Treg偏移,使其逐渐趋向免疫平衡,并调节血清中相关的炎症因子防止炎症的发生发展。

Objective To observe the effect of natural moxibustion at"Feishu(BL 13)"on immune balance of T helper cell 17(Th17)/regulatory T cell(Treg)in healthy rats.Methods Forty-eight rats were randomly divided into 10 rats in the sham moxibustion group and 38 rats in natural moxibustion group.The rats in the sham moxibustion group applied blank acupoint stickers to bilateral"Feishu(BL 13)",and the rats in natural moxibustion group applied acupoint stickers filled with Compound Banmao Ointment(复方斑蝥膏)to bilateral"Feishu(BL 13)"for a period of 8 h.Thirty rats in natural moxibustion group were successfully blistered after 8 h,and then were randomly divided into 1-day,3-day and 7-day groups with 10 rats in each group.The general condition of rats was recorded during the experiment;different time groups of natural moxibustion were sampled at the corresponding time,and HE staining was used to observe the pathological changes of the skin in the area of application;flow cytometry was used to detect the subpopulations of Th17 and Treg in peripheral blood,and the value of Th17/Treg was calculated;and ELISA was used to detect the serum interleukin 17A(IL-17A)and interleukin 6(IL-6),interleukin 10(IL-10)levels.Results Compared with sham moxibustion group,blisters can be seen in the application area of rats in 1-day natural moxibustion group,and the rats often scratched the skin of the moxibustion area,which showed loose stratum corneum,thickening of the stratum spinosum and stratum granulosum,absence of cells in the basal layer,inflammatory cell infiltration,and elevation of Treg in peripheral blood,and serum IL-17A,IL-6;in 3-day natural moxibustion group,the moxibustion area of the rats was scabbed and partially detached,with the most obvious dermatopathological changes,and elevated peripheral blood Th17 and Treg,and serum IL-17A,IL-6,IL-10;in 7-day natural moxibustion group,skin damage and pathological changes in the area of moxibustion were basically restored;Th17/Treg values were reduced in 1-,3-and 7-day moxibustion groups after blistering(P<0.05 or P<0.01).Compared with the 1-day moxibustion group,peripheral blood Th17,Treg,and serum IL-17A elevated in the 3-day moxibustion group;peripheral blood Treg,serum IL-17A,and IL-6 decreased,and Th17/Treg values elevated in the 7-day moxibustion group(P<0.05 or P<0.01).Compared with the 3-day moxibustion group,the 7-day moxibustion group had lower peripheral blood Th17 and Treg,serum IL-17A,IL-6,and IL-10,and higher Th17/Treg values(P<0.05 or P<0.01).Conclusion Natural moxibustion at"Feishu(BL 13)"can shift the Th17/Treg balance towards Treg of healthy rats,which will gradually lead to immune homeostasis,and regulate the relevant inflammatory factors in the serum to prevent inflammation from occurring and developing.