肺虚痰阻证模型大鼠肺组织水通道蛋白1 和5 分子表达与补肺化痰方对 其影响*

目的:1)从肺泡上皮水主动转运功能的角度探讨肺虚痰阻证的发生机理。2)通过观察肺虚痰阻证模型的AQP的活性及其相关基因、蛋白的表达和补肺化痰中药复方治疗前、后的对比,观察这一过程中上述指标的变化情况。方法:将雄性SD大鼠随机分为正常组、模型组、中药治疗组。模型组和治疗组造模40天,治疗组在造模26天后,药物灌胃治疗2周。采用组织化学染色法,对大鼠肺进行病理分析;RT-PCR的方法检测大鼠肺组织中AQP1、AQP5基因表达;western blot法检测大鼠肺组织中AQP1、AQP5蛋白水平。结果:1)与正常组相比,模型组局部出现明显炎症反应(P<0.01),治疗组局部炎症反应减轻(P<0.05)。2)mRNA结果显示,AQP1在正常组有表达,在模型组和治疗组未见表达。AQP5模型组与正常组相比,表达量显著增高(P<0.01);治疗组与模型组比较,表达量显著降低(P<0.01),但与正常组无显著差异。3)蛋白水平上,AQP1在模型组和治疗组与正常组相比差异显著(P<0.05),表达下降。AQP5模型组与正常组相比,显著升高(P<0.01);治疗组与模型组比较,显著下调(P<0.05);正常组表达低于治疗组,差异显著(P<0.05)。结论:1)AQP1和5基因及蛋白表达量变化是肺虚痰阻证的病理机制之一。2)补肺化痰中药复方可调节肺虚痰阻证模型大鼠肺组织AQP 5基因及蛋白表达。提示补肺化痰中药复方治疗肺虚痰阻证其作用机制与调节AQP5有关。

Effects of Compound Herbal Formulary on Expression of Aquaporin 1 and Aquaporin 5 in the Model of Rats with Phlegm Obstruction Due to Lung-Deficiency Syndromes

Objective: 1)To investigate the formative mechanism of phlegm obstruction due to lung-deficiency syndromes.2) To investigate the dynamic changes of the above parameters by detecting the effects of the herbal compound that "invigorates the lung and promotes expectorant" on the expression of AQP1 and AQP5 in the phlegm obstruction due to lung-deficiency syndromes model rats.Methods: Male SD Rats were randomly divided into three groups: normal,model,and treatment(with herbal Rats were treated by "invig-orate lung expectorant" herbal compound).The pathologic changes of lung were analyzed by microscopy with H&E staining.The mRNA and protein expressions of AQP1 and AQP5 were assayed by semi-quantitative RT-PCR and western blot respectively.Results: There was no expression of AQP1 mRNAs in the model and treated groups,while the expression of AQP5 mRNAs increased in the model group.Western blot analysis showed similar patterns of AQP5 protein being upregulated(P<0.01) while AQP1 downregulated(P<0.05).The expression of the AQP5 protein in the treated group decreased compared with that in the normal group(P<0.05).Conclusions: 1) One of the pathological mechanisms of the syndrome of "phelgm obstuction due to lung deficiency" is caused by the up-regulation of AQP5 and down-regulation of AQP1 gene and protein expression in lungs of the alveolar epithelium of rats.2) The expectorant herbal compound treatment can regulate the gene and protein expression of AQP5 but can’t regulate the AQP1 when the rats have this desease.These results suggest that the mechanism of this compound acting on this desease is related to the regulation of the expression of AQP1 AQP5.