阳和平喘颗粒对气道平滑肌细胞增殖的影响

Effect of Yanghepingchuan granules on proliferation of airway smooth muscle cells

目的:研究阳和平喘颗粒在体外对支气管哮喘大鼠气道平滑肌增殖及PI3K/PKB通路相关因子的影响。方法:将阳和平喘颗粒含药血清及PI3K/PKB通路阻断剂分别组合作用于体外培养的支气管哮喘大鼠气道平滑肌细胞,噻唑蓝比色法(MTT)检测大鼠气道平滑肌细胞的增值情况,实时聚合酶链反应(Realtime-PCR)检测阳和平喘颗粒对磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(PKB)、肾上腺素受体(AR)、内皮型一氧化氮合酶(e NOS)、增殖细胞核抗原(PCNA)mRNA表达水平的影响,蛋白印迹法(Western-Blot)检测阳和平喘颗粒对PI3K、PKB、AR、eNOS、PCNA蛋白表达水平的影响,酶联免疫吸附测定(ELISA)阳和平喘颗粒对磷脂酰肌醇二磷酸(PIP2)和磷脂酰肌醇三磷酸(PIP3)表达水平的影响。结果:50%阳和平喘颗粒(18.45g/kg)含药血清对支气管哮喘大鼠气道平滑肌细胞的增殖具有明显的抑制作用,阳和平喘颗粒含药血清及PI3K/PKB通路阻断剂能显著抑制支气管哮喘大鼠细胞内PI3K、PKB、PIP2、PIP3和PCNA的表达,促进eNOS和AR的生成。结论:阳和平喘颗粒具有在体外通过PI3K/PKB通路治疗支气管哮喘大鼠气道重塑的作用。

Objective： To study the effect of Yanghepingchuan Granule on airway smooth muscle proliferation and PI3 K/PKB pathway-related factors in bronchial asthma rats. Methods： The serum containing the optimal concentration of Yanghepingchuan granules and PI3 K/PKB pathway blockers were used to treat airway smooth muscle cells in bronchial asthma rats,the proliferation of rat airway smooth muscle cells was detected by MTT assay,the effect of Yanghepingchuan granules on expressions of phosphatidylinositol 3 kinase（ PI3 K）,protein kinase B（ PKB）,adrenergic receptor（ AR）,endothelial nitric oxide synthase（ e NOS） and proliferating cell nuclear antigen（ PCNA） was detected by Real time polymerase chain reaction（ Realtime-PCR）,expressions of phosphatidylinositol diphosphate（ PIP2） and phosphatidylinositol triphosphate（ PIP3） were detected by enzyme-linked immunosorbent assay（ ELISA）. Results： The serum containing Yanghepingchuan granule had obvious inhibitory effect on the proliferation of airway smooth muscle cells in bronchial asthma rats,expressions of PI3 K,PKB,PIP2,PIP3 and PCNA in bronchial asthma rats were significantly inhibited by the serum containing Yanghepingchuan granule and PI3 K/PKB pathway blockers,it promoted the generation of e NOS and AR. Conclusion： Yanghepingchuan granule has the effect on airway remodeling in bronchial asthma rats in vitro through the PI3 K/PKB signal pathway.