择时艾灸对类风湿性关节炎大鼠下丘脑-垂体-肾上腺轴昼夜节律的影响

目的:以中医针灸理论为指导,结合"子午流注"思想,运用分子生物学技术及生物节律分析方法,探讨择时艾灸对类风湿性关节炎(RA)大鼠下丘脑-垂体-肾上腺轴(HPAA)昼夜节律的影响机制。方法:SD大鼠随机分为空白组、模型组、艾灸组、假摘除肾上腺组(假摘组)、摘除肾上腺组(摘除组)、摘除肾上腺艾灸组(摘灸组),各组分4个时间点,分别为0点组、6点组、12点组、18点组,每个时间点6只大鼠。采用右后足足垫部皮内注入弗氏完全佐剂建立RA模型。造模后对摘除组、摘灸组大鼠进行摘除双侧肾上腺预处理。艾灸组、摘灸组大鼠接受麦粒灸"肾俞""足三里"治疗,每穴5壮,每日1次,6d为一疗程,共治疗3个疗程。各组大鼠在造模前、造模后、治疗后分别进行足容积测定。酶联免疫法检测血浆皮质酮(CORT)、促肾上腺皮质激素(ACTH)、促肾上腺皮质激素释放因子(CRF)的含量。Halbeg余弦法进行昼夜节律分析。结果:与空白组比较,模型组大鼠右后足肿胀度明显升高(P0.05),HPAA各指标保持昼夜节律性(P0.001,P0.05),峰相位有前移趋势,振幅有增高趋势,CORT中值明显降低(P0.05),其他HPAA指标中值明显增高(P0.05);与模型组比较,艾灸组大鼠右后足肿胀度明显降低(P0.05),HPAA各指标存在昼夜节律性(P0.001,P0.05),峰相位有后移趋势,振幅有降低趋势,CORT中值明显升高(P0.05),其他HPAA指标中值明显降低(P0.05);与艾灸组比较,摘灸组大鼠右后足肿胀度明显升高(P0.05),CORT和CRF昼夜节律消失,峰相位有前移趋势,振幅有降低趋势,CORT中值明显降低(P0.05),其他HPAA指标中值明显升高(P0.05)。结论:RA大鼠HPAA各指标具有明显的昼夜节律特征;艾灸可能通过调整RA大鼠HPAA各指标节律,达到控制炎性反应的作用。

Effects of Chronological Moxibustion on Circadian Rhythm Activities of Hypothalamus-Pituitary-Axis in Rheumatoid Arthritis Rats

Objective To observe the effect of chronological moxibustion on plasma corticosterone(CORT),corticotrophin releasing factor(CRF)and adrenocorticotropic hormone(ACTH)levels in rheumatoid arthritis(RA)rats,so as to explore its mechanisms underlying improvement of RA.Methods A total of 144 SD rats were randomly divided into normal control,model,moxibustion,sham-adrenalectomy(ADX,sham-ADX),ADX and ADX+moxibustion groups which were further separately divided into 4subgroups(4time-points:12pm,6am,12 am,6pm,n=6/subgroup).The RA model was established by subcutaneous injection of Freund's complete adjuvant(FCA,0.1mL)into the right footpad(for rats of the latter 5groups).In rats of the ADX and ADX+moxibustion groups,bilateral adrenal glands were removed under anesthesia.Moxibustion was applied to unilateral"Shenshu"(BL 23)and"Zusanli"(ST 36)for 10 min,once daily for 6days in a week,and continuously for 3weeks.The paw swelling volume was measured and plasma CORT,ACTH and CRF contents were assayed by ELISA and the circadian rhythm was analyzed by cosine curve fitting(cosinor)method.Results The paw swelling volume was significantly increased in the model group than in the normal control group(P<0.05),and decreased remarkably in the moxibustion group(P<0.05).The paw volume was markedly increased in the ADX+moxibustion group in comparison with the moxibustion group(P<0.05).In intact rats,plasma CRF and ACTH contents were significantly increased(P<0.05),and plasma CORT was obviously decreased in the model group(P<0.05).After moxibustion,the increased plasma CRF and ACTH contents and the decreased plasma CORT level were considerably reversed(P<0.05).In ADX rats,plasma CRF,ACTH and CORT contents of ADX and ADX+moxibustion groups had no significant changes compared with the sham-ADX group(P>0.05).Results of the cosine curve fitting analysis showed that the peak phases of plasma CORT contents were-355.78°at about 23:43in the normal control group,-309.05°at about 20:36in the model group,-326.50°at about 21:46in the moxibustion group,-291.65°at about 19:27in the ADX group and-300.87°at about 20:31in the ADX+moxibustion group.The peak phases of plasma ACTH contents were-324.08°at about 21:37in the normal control group,-295.39°at about 19:41in the model group,-310.81°at about 20:43in the moxibustion group,-146.51°at about 9:46in the ADX group and-267.64°at about 17:50in the ADX+moxibustion group.The peak phases of plasma CRF contents were-257.47°at about 17:10in the normal control group,-184.74°at about 12:19in the model group,-263.00°at about 17:32in the moxibustion group,-202.46°at about 13:30in the ADX group and-232.84°at about 15:31in the ADX+moxibustion group.It suggests that in the intact RA rats,moxibustion intervention may inhibit modeling-induced decrease of the circadian rhythm of plasma CORT,and hyperactivity of ACTH and CRF circadian rhythm.ConclusionMoxibustion intervention can relieve paw swelling in RA rats and modulate the circadian rhythm activities of plasma CORT,ACTH and CRF levels(HPA axis activities)in intact animals.