“邵氏五针法”配合双侧“足三里”对慢性哮喘模型大鼠肺组织中SP、VIP蛋白表达影响的研究

Effect of'Shao′s Five-Needle Therapy'Combined with Needling ST36 on Protein Expressions of SP and VIP in Lung Tissues of Rats with Chronic Asthma

目的:观察针灸“肺俞”“大椎”“风门”“足三里”对慢性哮喘模型大鼠行为体征、肺组织病理形态以及肺组织中P物质(SP)和血管活性肠肽(VIP)蛋白表达的影响,探讨其治疗慢性哮喘的作用机制。方法:将40只SPF级SD大鼠随机分为空白组、模型组、针刺组和艾灸组。除空白组外,其余各组采用雾化吸入卵蛋白致敏及激发方法制备哮喘模型,空白组给予生理盐水。造模成功后,针刺组、艾灸组大鼠均取“肺俞”(双)、“大椎”“风门”(双)、“足三里”(双)分别进行针刺和艾灸治疗,治疗后进行激发试验;空白组和模型组给予相同力度抓握刺激,不做其他治疗。隔日治疗1次,连续治疗6周。观察各组大鼠行为体征,末次治疗后摘取大鼠肺组织,苏木精-伊红(HE)染色观察大鼠肺组织病理形态,免疫组化染色观察各组大鼠肺组织中SP、VIP蛋白的表达。结果:空白组大鼠精神状态良好;模型组大鼠出现呛咳、喷嚏、呼吸急促,抓耳挠腮,烦躁不安等情况;与模型组相比,针刺组、艾灸组大鼠呼吸较平稳,反应灵敏。空白组大鼠肺组织结构完整、清晰,肺间质内未见明显炎性细胞浸润;模型组大鼠肺组织局部肺泡结构紊乱,支气管管腔内及其周围大量炎性细胞浸润,管腔狭窄;针刺组、艾灸组大鼠肺泡壁基本完整,支气管平滑肌轻度增厚,支气管管腔内及其周围少量炎性细胞浸润。模型组大鼠肺组织中SP蛋白表达明显高于空白组(P<0.01),VIP蛋白表达明显低于空白组(P<0.01);针刺组、艾灸组大鼠肺组织中SP蛋白表达明显低于模型组(P<0.01),VIP蛋白表达明显高于模型组(P<0.01)。结论:针灸“肺俞”“大椎”“风门”“足三里”可以通过下调肺组织中SP蛋白表达、上调肺组织中VIP蛋白表达改善肺组织炎症,这可能是“邵氏五针法”治疗哮喘的途径之一。

Objective:To observe the effect of needling Feishu(BL13),Dazhui(DU14),Fengmen(BL12)and Zusanli(ST36)on the expressions of substance P(SP)and vasoactive intestinal peptide(VIP)in lung tissues of rats with chronic asthma,thus to explore its action mechanism in the treatment of chronic asthma.Methods:40 SPF SD rats were randomly divided into the blank group,the model group,the acupuncture group and the moxibustion group.The asthma model was prepared by ovalbumin sensitization and stimulation.The acupuncture group was treated with needling BL13,DU14,BL12 and ST36,which were given moxibustion in the moxibustion group;both groups were followed by ovalbumin sensitization after the treatment.The blank group and the model group were stimulated by grasping without any other treatment.The treatment was once every other day for six weeks.The behavioral signs of rats in each group were observed.After the last treatment,the lung tissues of rats were collected.The pathological morphology of lung tissues was observed by HE staining,and the expressions of SP and VIP in lung tissues were observed by immunohistochemistry.Results:The rats in the blank group were in good state.The rats in the model group had symptoms of cough,sneeze,shortness of breath,scratching ears and cheeks,as well as irritability.Compared to those in the model group,the rats in the acupuncture group and in the moxibustion group had more stable breathing and fast reaction.In the blank group,the lung tissue structure was complete and clear,and no obvious inflammatory cell infiltration was found in the lung interstitium.In the model group,the local alveolar structure was in disorder,a large number of inflammatory cells infiltrated in and around the bronchial lumen,and the lumen was narrow.In the acupuncture group and in the moxibustion group,the alveolar walls were basically intact,the bronchial smooth muscles were slightly thickened,and a small amount of inflammatory cells infiltrated in and around the bronchial lumen.The expression of SP protein in lung tissues was significantly higher(P<0.01),and the expression of VIP protein was significantly lower in the model group than those in the blank group(P<0.01).The expression of SP protein in lung tissues was significantly lower(P<0.01),and the expression of VIP protein was significantly higher in the acupuncture group and in the moxibustion group than those in the model group(P<0.01).Conclusion:Needling BL13,DU14,BL12 and ST36 can improve inflammation of lung tissues by down-regulating the protein expression of SP and up-regulating the protein expression of VIP in lung tissues,which may be one of the mechanisms of'Shao's five-needle therapy'in the treatment of asthma.