穴位贴敷对支气管哮喘小鼠皮肤、肺组织TRPV1表达及IFN-γ、IL-4水平的影响

Effects of acupoint application on TRPV1 expression and IFN-γand IL-4 levels on skin and lung tissues of mice with bronchial asthma

目的观察穴位贴敷对支气管哮喘(bronchial asthma,BA)小鼠皮肤、肺组织中瞬时受体电位阳离子通道香草酸亚家族成员1(TRP cation channel subfamily V member 1,TRPV1)表达及血清中IFN-γ、IL-4水平的影响,探讨穴位贴敷治疗哮喘的机制。方法将75只BALB/c小鼠按随机数字表法分为空白组、模型组、穴位贴敷组、假贴敷组、地塞米松组,每组15只。采用屋尘螨滴鼻法制备BA小鼠模型。造模结束后,取小鼠双侧"肺俞""心俞""膈俞"进行穴位贴敷,穴位贴敷组贴敷药物为芥子、延胡索、细辛、甘遂药物组方,假贴敷组给予凡士林贴敷,1次/d,共贴敷14次;地塞米松组灌胃10 g/kg地塞米松,1次/d,连续干预14 d。运用EMKA-WBP肺功能检测系统检测小鼠气道阻力;天狼星红染色法、HE染色法观察小鼠肺组织病理形态改变,免疫组化法检测肺组织、皮肤组织中TRPV1表达,ELASA法检测肺泡灌洗液中IFN-γ、IL-4水平。结果在浓度为12.5、25.0、50.0 mg/ml的氯化乙酰甲胆碱溶液激发后,与模型组比较,穴位贴敷组、地塞米松组小鼠气道高阻力Penh值下降(P<0.05);肺组织HE染色和天狼猩红染色显示气管内炎性浸润和胶原纤维增生情况改善(P<0.05);与模型组比较,穴位贴敷组、地塞米松组肺组织TRPV1[(0.28±0.06)、(0.28±0.05)比(0.31±0.08)]表达降低(P<0.05);肺泡灌洗液中IFN-γ、IL-4水平降低(P<0.05)。结论穴位贴敷可通过诱发皮肤反应治疗哮喘,其作用机制可能与激活肺组织、皮肤组织中TRPV1,降低IFN-γ、IL-4水平有关。

Objective To observe the effects of acupoint application on TRPV1 expression on skin and lung tissue and levels of serum IFN-γ and IL-4 of mice with bronchial asthma,and to explore the mechanism of acupoint application in the treatment of asthma.Methods 75 BALB/C mice were divided into control group,model group,acupoint application group,sham application group and dexamethasone group,15 in each group.The bronchial asthma model of mice was prepared by nasal drop method of Dermatophagoides pteronyssinuus(HDM).After the modeling,the acupoints such as"Feishu","Xinshu"and"Geshu"of each side of the mice were taken for application.The acupoint application group was treated with white mustard seed,Corydalis yanhusuo,asarum and Gansui group,while the sham application group was treated with vaseline,once a day,14 times in total.Emka-wbp pulmonary function detection system was used to detect airway resistance in mice.The expression of TRPV1 on lung tissues and skin tissues were detected by immunohistochemical method.The levels of IFN-γ and IL-4 were detected by ELASA method.Results After the stimulation of methacholine chloride solution at the concentrations of 12.5,25.0 and 50.0 mg/ml,compared with the model group,the Penh value(airway high resistance)of mice in the acupoint application group and dexamethasone group decreased(P<0.05).HE staining and Sirius red staining of lung tissue showed that inflammatory infiltration and collagen fiber proliferation had been improved(P<0.05).Compared with model group,the expression of TRPV1(0.28±0.06,0.28±0.05 vs.0.31±0.08)in lung tissues of acupoint application group and dexamethasone group significantly decreased(P<0.05).The levels of IFN-γ and IL-4 in BALF decreased(P<0.05).Conclusion The mechanism of skin reaction induced by acupoint application in treating asthma may be related to the changes of IFN-γ,IL-4 and other related factors caused by the activation of TRPV1 in lung tissues and skin tissues.