降气化痰推拿法对慢性哮喘幼鼠气道重塑及TGF-β1、Smad3表达的影响

Effect of Qi-depressing and phlegm-eliminating massage on airway remodeling and expression of transforming growth factorβ1 and recombinant SMAD family member 3 in young rats with chronic asthma

目的探讨降气化痰推拿法对慢性哮喘幼鼠气道重塑的治疗作用及潜在机制。方法将50只SPF级雌性BALB/c小鼠随机分为对照组(A组)、模型组(B组)、推拿组(C组)、地塞米松组(D组)、推拿+地塞米松组(E组),每组10只。其中除A组外,其余小鼠均制备哮喘模型。第16天开始,C、D、E组进行相应推拿、地塞米松、推拿+地塞米松治疗,B组注射等量生理盐水;第22天开始,各组连续激发7 d。比较各组的肺功能、血清白介素4(interleukin 4,IL-4)水平、肺组织病理改变以及肺组织转化生长因子β1(TGF-β1)、Smad3 mRNA和蛋白表达水平。结果肺能功方面,与A组相比,各组给药后的气道狭窄指数明显升高、肺顺应性下降,且C、D、E组的气道狭窄指数低于B组,肺顺应性高于B组。在高浓度乙酰胆碱(6.250 g·L^(-1),12.500 g·L^(-1),25.000 g·L^(-1))给药时,E组的气道狭窄指数明显低于C、D组,且C组低于D组,C组肺顺应性低于D组(P<0.05);炎性水平方面,与A组相比,各组IL-4水平、炎症细胞浸润程度均升高,且C、D、E组均低于B组,E组低于C、D组,差异具有统计学意义(P<0.05);HE染色结果显示,A组肺组织无明显炎性改变,B组炎性细胞浸润严重,C组、D组、E组轻微炎症浸润,且E组浸润程度低于C、D组;与A组相比,各组气道壁厚度和气道平滑肌厚度升高,且、D、E组均低于B组,E组低于C、D组;与A组相比,各组TGF-1β、Smad3 mRNA和蛋白水平均升高,且C、D、E组均低于B组,E组低于C、D组,差异具有统计学意义(P<0.05)。结论降气化痰推拿法能够有效改善慢性哮喘幼鼠的气道重塑,降低炎性反应水平,减轻炎性浸润,其机制可能与下调TGF-β1/Smad3通路表达相关。

Objective To explore the therapeutic effect and potential mechanism of Qi-depressing and phlegm-eliminating massage on airway remodeling in young rats with chronic asthma.Methods 50 SPF grade female BALB/c mice were randomly divided into the control group(group A),the model group(group B),the massage group(group C),the dexamethasone group(group D)and the massage+dexamethasone group(group E),with 10 mice in each group.Except group A,the other mice were all prepared with asthma models.From the 16th day,group C,group D and group E were treated with corresponding massage,dexamethasone,and massage+dexamethasone,while group B was injected with the equal amount of normal saline.From the 22nd day,each group was stimulated continuously for 7 days.Pulmonary function,serum interleukin-4(IL-4)level,pulmonary tissue pathological changes,and transforming growth factorβ1(TGF-β1)level,Smad3 mRNA level and protein expression levels in lung tissues in each group were compared.Results In terms of pulmonary function,compared with group A,the airway stenosis index was significantly higher and lung compliance was lower in each group after administration.The airway stenosis index of group C,group D and group E was lower than that of group B,and lung compliance was higher than that of group B.When high concentrations of acetylcholine(6.250 g·L^(-1),12.500 g·L^(-1),25.000 g·L^(-1))were administered,the airway stenosis index of group E was significantly lower than that of group C and group D,and group D was lower than group C,and the lung compliance in group C was lower than that in group D(P<0.05).In terms of inflammatory levels,compared with group A,the level of IL-4 and the degree of infiltration of inflammatory cells in each group increased,and group C,group D and group E were lower than group Group B,while Group E was lower than Groups C and group D,and there was statistically significant difference(P<0.05).HE staining results showed that there were no obvious inflammatory changes in the lung tissues of group A,group B had severe inflammatory cell infiltration,and group C and group D and group E had mild inflammatory infiltration,and the degree of infiltration in group E was lower than that in group C and group D.Compared with group A,the thickness of the airway wall and the thickness of the airway smooth muscle in each group increased,and group D and group E were lower than group B,and group E was lower than group C and group D.Compared with group A,TGF-1βlevel and Smad3 mRNA level and protein expression levels increased in each groups,and group C,group D and group E were lower than group B,and group E was lower than group C and group D,and there was statistically significant difference(P<0.05).Conclusion Qi-depressing and phlegm-eliminating massage can effectively improve airway remodeling,reduce inflammatory response level,and alleviate inflammatory infiltration in young rats with chronic asthma.And its mechanism may be related to the down-regulation of TGF-β1/Smad3 pathway expression.