木姜子和忍冬藤配伍醇提物影响哮喘小鼠ASMC增殖与气道重塑的效果

Effect of ethanol extracts of Litsea and Lonicera japonica Thunb on ASMC proliferation and airway remodeling in mice with asthma

目的观察木姜子和忍冬藤配伍醇提物减轻哮喘小鼠气道平滑肌细胞增殖的作用及探讨其可能的起效机制。方法将健康清洁级昆明种小鼠随机分为空白组,模型组,木姜子和忍冬藤配伍醇提物低、中、高剂量组5组,每组10只。模型组,木姜子和忍冬藤配伍醇提物低、中、高剂量组采用卵清蛋白腹腔注射致敏+雾化吸入激发法建立哮喘小鼠模型。木姜子和忍冬藤配伍低、中、高剂量组每天给予木姜子和忍冬藤的醇提物(剂量分别为5 mg/kg、10 mg/kg、20 mg/kg)灌胃,其余各组给予等量的生理盐水,均是每天1次,连续干预7 d。干预结束后,ELISA法检测血清转化生长因子-β1(TGF-β1)、白细胞介素-4(IL-4)、白细胞介素-13(IL-13)、血管内皮生长因子(VEGF)、血小板源生长因子(PDGF)水平,HE染色观察肺组织形态学变化,Western blot法检测细胞周期蛋白1(Cyclin D1)、细胞外信号调节激酶(ERK1/2)蛋白表达水平。结果与空白组比较,模型组TGF-β1、IL-4、IL-13、VEGF、PDGF水平及Cyclin D1、ERK1/2蛋白表达水平均升高,差异均有统计学意义(P<0.05或P<0.01);与模型组比较,低、中、高各剂量组TGF-β1、IL-4、IL-13、VEGF、PDGF水平及Cyclin D1、ERK1/2蛋白表达水平均降低,差异均有统计学意义(P<0.05或P<0.01);肺组织形态学变化显示各剂量组能够改善肺组织细胞炎性浸润病理变化。结论木姜子和忍冬藤配伍醇提物可降低TGF-β1、IL-4、IL-13、VEGF、PDGF水平及Cyclin D1、ERK1/2蛋白表达水平,其起效机制与抑制气道平滑肌细胞的增殖有关。

Objective To observe the effect of ethanol extracts of Litsea and Lonicera japonica Thunb on proliferation of airway smooth muscle cells(ASMC)in mice with asthma and to discuss the possible effective mechanism.Methods Healthy and clean Kunming mice were randomly divided into a blank group,a model group,the low dose,middle dose,high dose groups of ethanol extracts of Litsea and Lonicera japonica Thunb,with 10 mice in each group.The asthmatic mouse models were established by intraperitoneal injection of ovalbumin and by stimulation of inhalation.The mouse in the low-dose,medium-dose and high-dose groups were intragastrically administrated with ethanol extracts of Litsea and Lonicera japonica Thunb(5 mg/kg,10 mg/kg,20 mg/kg,respectively),while the other groups were given the same amount of normal saline once a day for consecutive 7 days.After the intervention,ELISA was adopted to detect the levels of transforming growth factor-β1(TGF-β1),interleukin-4(IL-4),interleukin-13(IL-13),vascular endothelial growth factor(VEGF),platelet-derived growth factor(PDGF)in serum.HE staining was used to observe the morphological changes of lung tissue.The protein expression levels of Cyclin D1 and extracellular signal-regulated kinase(ERK1/2)were detected by Western blot.Results Compared with the blank group,the model group had increased levels of TGF-β1,IL-4,IL-13,VEGF,PDGF as well as the protein expressions of Cyclin D1 and ERK1/2,and the differences were statistically significant(P<0.05 or P<0.01).Compared with the model group,the low,medium and high dose groups had decreased levels of TGF-β1,IL-4,IL-13,VEGF,PDGF and the protein expressions of Cyclin D1 and ERK1/2,and the differences were statistically significant(P<0.05 or P<0.01).The morphological changes of lung tissue showed that each dose group could improve the pathological changes of inflammatory infiltration of lung tissue cells.Conclusion Ethanol extracts of Litsea and Lonicera japonica Thunb can decrease the levels of TGF-β1,IL-4,IL-13,VEGF,PDGF and the protein expressions of Cyclin D1 and ERK1/2,and the mechanism is related to the inhibition of proliferation of airway smooth muscle cell.