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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。
OBJECTIVE: 1) To explore the pathogenesis of pulmonary phlegm obstruction from the perspective of active transport of alveolar epithelial water. 2) Observe the changes of AQP activity, the expression of related genes and proteins and the TCM compound prescription of Bufei Huatan and phlegm before and after treatment by observing the changes of AQP. Methods: Male SD rats were randomly divided into normal group, model group and TCM group. Model group and treatment group modeling 40 days, the treatment group after 26 days in the model, the drug gavage for 2 weeks. The histopathology of rat lungs was analyzed by histochemical staining. The expression of AQP1 and AQP5 genes in rat lungs was detected by RT-PCR. The expression of AQP1 and AQP5 in lungs was detected by western blot. Results: 1) Compared with the normal group, there was a significant inflammatory reaction in the model group (P <0.01), and the local inflammatory response in the treatment group was alleviated (P <0.05). 2) mRNA results showed that AQP1 expression in normal group, the model group and the treatment group no expression. The expression of AQP5 in model group was significantly higher than that in normal group (P <0.01). The expression of AQP5 in model group was significantly lower than that in model group (P <0.01), but not significantly different from that in normal group. 3) protein level, AQP1 in the model group and the treatment group compared with the normal group significant difference (P <0.05), the expression decreased. Compared with the normal group, the AQP5 model group was significantly higher (P <0.01); the treatment group was significantly lower than the model group (P <0.05); the normal group was lower than the treatment group, the difference was significant (P <0.05). Conclusion: 1) The changes of AQP1 and 5 gene and protein expression are one of the pathological mechanisms of lung phlegm blocking syndrome. 2) Bufei Huatan Chinese medicine compound can regulate the expression of AQP 5 gene and protein in lung tissue of rats with lung-phlegm-dampness syndrome. Promote the lungs and phlegm Chinese medicine compound treatment of lung phlegm block its mechanism of action and regulation AQP5.