P物质对支气管哮喘大鼠气道重塑及自噬-凋亡的影响  

Effects of Substance P on Airway Remodeling and Autophagy Apoptosis in Asthmatic Rats

作　　者：王心晓[1] 欧宗兴[1] 陈忠仁 王蕾 冯书 WANG Xin-xiao;OU Zong-xing;CHEN Zhong-ren;WANG Lei;FENG Shu(Department of Respiratory Medicine and Critical Care,Central South University Xiangya School of Medicine Affiliated Haikou Hospital,Haikou 570208,China)

机构地区：[1]中南大学湘雅医学院附属海口医院呼吸与危重症医学科,海南海口570208

出　　处：《药物生物技术》2021年第4期369-375,共7页Pharmaceutical Biotechnology

基　　金：海南省自然科学基金青年基金项目;海南省自然科学基金项目(No.819QN385)。

摘　　要：观察P物质(SP)对支气管哮喘大鼠气道重塑的影响,并探究其对细胞自噬-凋亡的调控机制。60只健康雌性SD大鼠随机分为6组(n=10):对照组(NC)、模型组(Model组)、SP组、SP拮抗组(SPi组)、自噬诱导组(Rapa组)和自噬抑制组(3-MA组),除NC组外其余5组以卵清蛋白雾化吸入法建立支气管哮喘模型,NC组用生理盐水代替卵清蛋白进行雾化吸入。NC组和Model组尾静脉注射生理盐水,SP组给予外源性SP,SPi组给予SP拮抗剂Aprepitant,Rapa组给予雷帕霉素,3-MA组给予3-MA,剂量均为0.1 mL/kg。观察各组乙酰胆碱激发后气道阻力,比较各组白细胞总数、嗜酸性粒细胞、支气管病理学改变和气道重塑(S/Pi、W/Pi、N/Pi)情况、ELISA法检测血清IgE水平,RT-PCR、Western blot检测支气管组织中Caspase-3、Beclin-1、Bcl-2和P62等表达。与NC组比较,Model组气道阻力显著升高(P<0.05),与Model组比较,SP组气道阻力升高,Spi组、Rapa组和3-MA组气道阻力降低;且随着雾化剂浓度增加,各组大鼠气道阻力升高(均P<0.05)。与Model组比较,SP组白细胞总数、嗜酸性粒细胞比例和IgE水平显著升高,Spi组、Rapa组和3-MA组白细胞总数、嗜酸性粒细胞比例和IgE水平显著降低(P<0.05)。与NC组比较,Model组大量炎性细胞浸润;与Model组比较,SP组改善不明显;Spi组、Rapa组和3-MA组细胞浸润,气道平滑肌增生,气道壁厚度、支气管狭窄程度显著改善。与NC组比较,Model组S/Pi、W/Pi和N/Pi显著升高;与Model组比较,SP组S/Pi、W/Pi和N/Pi显著升高,Spi组、Rapa作用和3-MA组S/Pi、W/Pi和N/Pi显著降低(P<0.05)。与NC组比较,Model组Caspase-3和Beclin-1 mRNA显著升高,Bcl-2和P62 mRNA显著降低;与Model比较,SP组和Rapa组Caspase-3和Beclin-1 mRNA显著升高,Bcl-2和P62 mRNA显著降低;Spi组和3-MA组Caspase-3和Beclin-1 mRNA显著降低,Bcl-2和P62 mRNA显著升高(P<0.05)。与NC组比较,Model组Caspase-3和Beclin-1蛋白显著升高,Bcl-2和P62蛋白显著降低;与Model�To observe the effect of substance P(SP)on airway remodeling in asthmatic rats and explore the regulatory mechanism on autophagy apoptosis,60 healthy female SD rats were randomly divided into 6 groups(n=10):control group(NC group),model group(Model group),SP group,SP antagonist group(Spi group),autophagy induction group(Rapa group)and autophagy inhibition group(3-MA group).Except the NC group,the other 5 groups were established bronchial asthma models by ovalbumin atomization inhalation,and the NC group was used normal saline instead of ovalbumin for atomization inhalation.NC group and Model group were injected with normal saline,SP group was given exogenous SP,Spi group was given SP antagonist apreptant,Rapa group was given rapamycin,3-MA group was given 3-MA,the dose were all 0.1 mL/kg.The airway resistance after acetylcholine challenge were observed,the total leukocyte count eosinophils,bronchial pathological changes and airway remodeling(S/Pi,W/Piand N/Pi)were compared.The level of serum IgE was detected by ELISA,the expression of Caspase-3,beclin-1,Bcl-2 and P62 in bronchial tissue were detected by RT-PCR and Western blot.Compared with NC group,the airway resistance in model group was increased significantly(P<0.05).Compared with Model group,the airway resistance in SP group was increased,while the airway resistance in Spi group,Rapa group and 3-MA group were decreased;and with the increase of aerosol concentration,the airway resistance were increased(P<0.05).Compared with Model group,the total number of leukocytes,eosinophil ratio and IgE level in SP group were significantly increased,while the Spi group、Rapa group and 3-MA group were significantly decreased(P<0.05).Compared with NC group,the Model group had have a large number of inflammatory cells infiltration,compared with Model group,SP groups showed no significant improvement or even worse than those in model group,and Spi group、Rapa group and 3-MA group,cell infiltration,airway smooth muscle hyperplasia,airway wall thickness and degree of bronchia

关 键 词：P物质 支气管哮喘 气道重塑 细胞自噬 细胞凋亡 

分 类 号：R22[医药卫生—中医基础理论] R285.5[医药卫生—中医学]