Sema4D在病毒诱发哮喘大鼠气道重塑中的作用及咳喘宁的干预

The Role of Sema4D in Airway Remodeling Induced by Virus in Asthmatic Rats and the Intervention of Kechuanning(咳喘宁)

目的:探讨脑信号蛋白4D(Semaphorins4D,Sema4D)对呼吸道合胞病毒(Respiratory syncytial virus,RSV)诱发哮喘大鼠气道重塑的影响及咳喘宁的改善作用。方法:构建RSV复合卵清白蛋白(Ovalbumin,OVA)模型,实验大鼠随机分为模型组、咳喘宁0.2 g/kg、2.1 g/kg、7 g/kg组、Sema4D抗体0.01 mg/kg组,另设正常对照组,灌胃予以相应药物或生理盐水干预,连续2 w。检测大鼠气道反应性,HE染色观察各组大鼠肺组织病理改变;Masson染色观察各组大鼠肺组织胶原沉积状况;免疫组织化学观察各组大鼠肺组织α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、基质金属蛋白酶-2(matrix metalloprotein-2,MMP-2)的表达;蛋白印迹试验检测肺组织中Sema4D的表达。结果:模型组大鼠气道反应性显著高于正常组大鼠,提示造模成功;与模型组相比,咳喘宁0.2 g/kg、2.1 g/kg、7 g/kg剂量组及Sema4D抗体0.01 mg/kg组胶原沉积明显减少,Sema4D、α-SMA、MMP-2蛋白表达降低,更接近正常组水平,其中以Sema4D抗体0.01 mg/kg组及咳喘宁7 g/kg组改善最为明显。结论:Sema4D可以影响α-SMA、MMP-2的表达及胶原沉积,从而产生气道重塑,而咳喘宁可能通过调节Sema4D改善气道重塑。

Objective:To investigate the effect of Sema4 D on airway remodeling induced by RSV in asthmatic rats and the improvement of Kechuanning.Methods:RSV combined with OVA model was established.The rats were randomly divided into the control group,the model group,0.2 g/kg,2.1 g/kg,7 g/kg Kechuanning groups,and 0.01 mg/kg Sema4 D antibody group,and given corresponding drug intervention or saline water for 2 weeks constantly.The airway responsiveness of rats was measured,HE staining was used to observe pathological changes of lung tissues in rats,the collagen deposition in the lung tissue was observed by Masson staining.Immunohistochemistry method was used to observe the expressions ofα-SMA and MMP-2 in lung tissues.Sema4 D expression in the lung tissue was detected by Western Blot.Results:Compared with the control group,the airway responsiveness in the model group was significantly increased,suggesting that the model was successful.Compared with the model group,the collagen depositions in 0.2 g/kg,2.1 g/kg,7 g/kg Kechuanning groups and 0.01 mg/kg Sema4 D antibody group were significantly decreased,and the expression levels of Sema4 D,α-SMA and MMP-2 were decreased more closely to those in the control group.The improvements in 0.01 mg/kg Sema4 D antibody group and 7 g/kg Kechuanning group were the most obvious.Conclusion:Sema4 D can affect the expressions ofα-SMA,MMP-2 and collagen deposition,thus produce airway remodeling.Kechuanning can improve airway remodeling by regulating Sema4 D.