逆灸对佐剂性关节炎大鼠下丘脑中枢性应激激素变化的影响

Preventive moxibustion in the changes of central stress hormone in hypothalami in rats with adjuvant arthritis

目的:观察逆灸对随后佐剂性关节炎大鼠原发足组织病理学变化及中枢性应激激素变化的影响。方法:实验在北京中医药大学针灸学院针灸机理实验室进行。以随机数字表法将48只Wistar大鼠分为6组,每组8只。①正常组:不造模,不预灸。②模型早期组:大鼠右后足掌皮下注射福氏完全佐剂0.1mL造成佐剂性关节炎模型,造模第3天取材。③逆灸早期组:于造模前进行艾灸,大椎穴处施灸,每次1壮,隔日灸1次,共灸8次,并于灸结束后造模,造模第3天取材。④模型继发组:造模,造模第16天取材。⑤逆灸继发组:于造模前进行艾灸,并于灸结束后造模,造模第16天取材。⑥逆灸正常组:正常大鼠进行艾灸,灸结束后取材。采用放射免疫法观察各组大鼠下丘脑促肾上腺皮质激素释放激素及β内啡肽含量,苏木精-伊红染色观察原发足组织病理学变化。结果:48只大鼠均进入结果分析。①苏木精-伊红染色结果:模型各组关节滑膜间有大量炎性细胞浸润,滑膜水肿;逆灸各组较模型组减轻。②下丘脑促肾上腺皮质激素释放激素水平:模型早期水平与模型继发组均较正常组高犤(541.65±74.95),(479.70±59.08),(187.14±42.24)ng/g,P<0.01犦;逆灸模型早期与继发组均较同期模型组低犤(365.04±69.87),(367.14±35.93)ng/g,P<0.05犦。③β内啡肽水平:模型早期组水平高于正常组,模型继发组与正常组水平相似犤(24.16±2.16),(15.86±1.37),(13.74±1.24)μg/g,P<0.01犦,逆灸早期组低于同期模型组,但逆灸继发组高于早期组犤(19.28±1.91),(21.75±1.68)μg/g,P<0.05犦。结论:逆灸、佐剂和逆灸后再接受佐剂3种情况对大鼠下丘脑中促肾上腺皮质激素释放激素、β内啡肽应激激素产生不同的影响。逆灸和佐剂刺激均可使大鼠下丘脑中促肾上腺皮质激素释放激素和β内啡肽的含量产生增高的趋势,但佐剂刺激比逆灸强烈得多,使大鼠下丘脑促肾上腺皮质激素释放激素在早期和继发期均异常升高,使大鼠下丘脑β内啡肽早期异常升高,但继发期开始下降到正常水平。逆灸、佐剂作为不同的应激原均可引起机体的应激反应,只是程度和延续时间存在差异。若将两种性质不同的应激互相前后叠落,即逆灸后再接受佐剂刺激,则呈现局部和全身的早期和继发期的病理应激反应的减轻综合效果,同时使早期,异常升高的促肾上腺皮质激素释放激素和β内啡肽开始下调;但到继发期时促肾上腺皮质激素释放激素仍然维持缓降的趋势,β内啡肽却又开始上升,显然两种激素在佐剂性关节炎发展的不同时段以及逆灸对其的影响呈现出并不完全相同的变化趋势。

AIM:To observe the effect of preventive moxibustion on the pathology and stress hormone in the primary foot tissue of rats with adjuvant arthritis,and investigate its protective mechanism.METHODS:The experiment was carried out in the laboratory of acupuncture mechanism,School of Acupuncture,Beijing University of Traditional Chinese Medicine from August to December 2004.Forty eight Wistar rats were divided into 6 groups with the method of randomly number table:normal group,model early group,preventive moxibustion early group,model secondary group,preventive moxibustion secondary group and preventive moxibustion normal group with 8 rats in each group.Rats in the normal group were not made into models and received no preventive moxibustion;Rats in the model early group were made into adjuvant arthritis models by using subcutaneous injection of Zhao's complete adjuvant(0.1 mL) at palm of right hindfoot,and the samples were prepared on the 3rd day;Rats in the preventive moxibustion early group were made into adjuvant arthritis models after muxibustion at Tachui acupoint once every other day,one moxa cone every time for 8 times,and the samples were prepared on the 3rd day;Rats in the model secondary group were made into adjuvant arthritis models,and the samples were prepared on the 16th day;Rats in the preventive moxibustion secondary group were made into adjuvant arthritis models after muxibustion,and the samples were prepared on the 16th day;Rats in the preventive moxibustion normal group were treated with moxibustion,and then the samples were prepared after that.The contents of hypothalamic corticotropin releasing hormone and beta endorphin of rats in each group were observed with radioimmunoassay,and the pathological changes of primary foot tissue were observed with haematine eosin staining.RESULTS:All the 48 rats were involved in the analysis of results.① The results of haematine eosin staining:Plenty of inflammatory infiltration between synovium of joint and synovial edema could be observed in each model group,and they were lighter in each preventive moxibustion groups than in the model group.② Level of hypothalamic corticotropin releasing hormone:It was higher in the model early group and model secondary group than in the normal group[(541.65± 74.95),(479.70± 59.08),(187.14± 42.24) ng/g,P< 0.01];It was lower in the preventive moxibustion early and secondary groups than in the model group at the same period[(365.04± 69.87), (367.14± 35.93) ng/g,P< 0.05].③ Level of beta endorphin:It was higher in the model early group than in the normal group,which was similar to that in the model secondary group[(24.16 ± 2.16),(15.86± 1.37),(13.74± 1.24) μ g/g,P< 0.01];It was lower in the preventive moxibustion early group than in the model group at the same period,but it was higher in the preventive moxibustion secondary group than in the preventive moxibustion early group[(19.28± 1.91),(21.75± 1.68) μ g/g,P< 0.05].CONCLUSION:Preventive moxibustion,adjuvant,and adjuvant after preventive moxibustion have different influences on hypothalamic corticotropin releasing hormone and beta endorphin of rats.The stimulations of both preventive moxibustion and adjuvant can increase the contents of hypothalamic corticotropin releasing hormone and beta endorphin of rats,but the stimulation of adjuvant is stronger,it leads to the abnormal increase of the level of hypothalamic corticotropin releasing hormone at early and secondary stage,and also the abnormal increase of hypothalamic beta endorphin at early stage,but it decreases to the normal level at secondary stage.As different stressors,both preventive moxibustion and adjuvant can result in the stress reaction of organ,but there are differences in the degree and lasting duration.If stressors of two different natures are overlapped to each other,that is,adjuvant is given after preventive moxibustion,the comprehensive effect of the relief of pathological stress reaction of local and whole body at early and secondary stages will occur,and down regulate the abnormally increased hypothalam