TIMP-1在哮喘大鼠气道重塑中的作用及抗RANTES抗体的干预影响
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摘要
背景:支气管哮喘(简称哮喘)是一种呼吸系统常见的慢性疾病,是以气道高反应性、气道慢性炎症及气道重塑为特征的变态反应性疾病。随着社会的发展,其发病率不断上升,特别是儿童哮喘更加明显,给社会及家庭带来长期的负担和压力。因此,哮喘的发病机制和治疗一直是研究的热点,人们对哮喘的认识已经开始在分子-基因水平上探究病因、发病机制及相应的诊断和治疗方法。但是,哮喘的发生是多因素、多基因、多环节相互作用的结果,许多问题有待解决,进一步研究哮喘的发病机制及寻求新的治疗方法成为当前医学界普遍关注和迫切解决的问题。
目的:以哮喘大鼠气道模型为基础,通过研究金属蛋白酶组织抑制剂-1(TIMP-1)在气道重塑中的表达情况,并用计算机图像分析软件对气道重塑程度情况进行量化,进而探讨哮喘大鼠模型中TIMP-1与气道重塑的关系;同时应用抗正常T细胞表达和分泌的活性调节蛋白抗体(抗RANTES抗体)对哮喘大鼠进行干预,通过肺组织中TIMP-1表达的变化以及量化的气道重塑情况改变,来探讨抗RANTES抗体对气道重塑的影响,从而为哮喘的临床治疗提供动物实验依据。
方法:将36只健康雄性wistar大鼠随机分为三组:哮喘模型组、抗RANTES抗体干预组、正常对照组,每组12只。哮喘模型组和抗RANTES抗体干预组于第1、7、14天腹腔注射10%抗原液1ml(含卵白蛋白100mg、氢氧化铝100mg)进行致敏,正常对照组腹腔注射等量生理盐水致敏。从第21天起将哮喘模型组、抗RANTES抗体干预组大鼠分别置于密闭玻璃罩内,雾化吸入1%的卵白蛋白激发,每次雾化吸入30分钟,隔天1次,共6周,正常对照组用等量的生理盐水进行激发。抗RANTES抗体干预组自雾化激发后1周起即从第28天起每次激发前半小时给予尾静脉注射抗RANTES抗体1ug,哮喘模型组和正常对照组给予尾静脉注射等量生理盐水,隔天1次,共5周。观察三组实验大鼠的一般状况,包括活动、呼吸、神态、毛发等。末次激发结束后24小时用1%戊巴比妥那0.2ml/只腹腔注射麻醉各组大鼠,打开胸腔,取出左肺组织。作HE染色病理图片,光镜观察三组实验大鼠的支气管肺组织的病理改变、用计算机图像分析软件分别测定三组实验大鼠支气管基底膜周长(Pbm)、气道平滑肌面积(WAm)、气道内壁面积(WAi),并用Pbm标准化,分别以WAm/Pbm及WAi/Pbm表示气道平滑肌面积、气道内壁面积,量化气道重塑程度的改变;采用免疫组化方法测定三组实验大鼠肺组织TIMP-1的相对积分光密度的改变,观察TIMP-1的表达情况。
结果:(1)三组大鼠的一般状况:①哮喘模型组:雾化吸入卵白蛋白后大部分大鼠出现烦躁不安,搔抓颜面,鼻扇,毛发失去光泽,打喷嚏,口唇发绀,继而大鼠安静不动,弓背,张口呼吸,呼吸加深加快,前肢缩抬,四肢颤抖,口鼻流出粘液,精神不振和反应迟钝,停止雾化后上述表现逐渐缓解。②抗RANTES抗体干预组:第1周每次雾化吸入卵白蛋白后,大部分表现与哮喘模型组相似,而在应用抗RANTES抗体后,虽有烦躁、喷嚏、四肢抓挠,但无明显的呼吸困难、口唇发绀表现。③正常对照组:致敏及任意一次激发后均正常饮食、饮水,精神活泼、反应灵敏、呼吸平稳、毛色光洁,没有呼吸急促、烦躁不安或口周发绀的表现。
(2)三组大鼠气道平滑肌的比较:哮喘模型组大鼠气道平滑肌面积WAm/Pbm[(5.78±1.36)μm2/μm]明显大于正常对照组大鼠的气道平滑肌面积WAm/Pbm[(3.35±1.01)μm2/μm] (P<0.05);抗RANTES抗体干预组大鼠气道平滑肌面积WAm/Pbm[(4.12±1.21)μm2/μm]较哮喘模型组大鼠的气道平滑肌面积WAm/Pbm显著降低(P<0.05);抗RANTES抗体干预组大鼠的气道平滑肌面积WAm/Pbm与正常对照组大鼠气道平滑肌面积WAm/Pbm相比无差别(P>0.05)。
(3)三组大鼠气道内壁面积的比较:哮喘模型组大鼠气道内壁面积WAi/Pbm[(15.10±3.99)μm2/μm]明显大于正常对照组大鼠的气道气道内壁面积WAi/Pbm[(9.25±2.29)μm2/μm] (P<0.05);抗RANTES抗体干预组大鼠气道内壁面积WAi/Pbm[(10.01±2.69)μm2/μm]较哮喘模型组大鼠的气道内壁面积WAi/Pbm显著降低;抗RANTES抗体干预组大鼠的气道内壁面积WAm/Pbm与正常对照组大鼠的气道内壁面积WAi/Pbm相比无差别(P>0.05)。
(4)三组实验大鼠肺组织TIMP-1表达的变化:哮喘模型组大鼠肺组织的TIMP-1表达[积分光密度(1.9580±0.0927)]较正常对照组大鼠肺组织的TIMP-1表达[积分光密度(0.7623±0.1000)]增强(P<0.05);哮喘模型组大鼠肺组织的TIMP-1表达较抗RANTES抗体干预组大鼠肺组织的TIMP-1表达[积分光密度(0.9770±0.0233)]增强(P<0.05);抗RANTES抗体干预组大鼠肺组织的TIMP-1表达与正常对照组大鼠肺组织的TIMP-1表达相比无差别(P>0.05)。
结论:(1)哮喘大鼠模型成功;(2)哮喘模型组的大鼠肺组织中TIMP-1的表达高,可能在气道重塑中起到了重要的作用;(3)哮喘大鼠在应用抗RANTES抗体后哮喘症状缓解,气道重塑程度减轻,通过体外干预研究证实抗RANTES抗体可抑制气道重塑,它可能延缓气道重塑的发生。
Background:Asthma is an allergic disease,one common disease in chronic illness of respiratory system.Its airway is characteristed by hyperreactivity,chronic inflammation and remodeling.With the development of society,the incidence of asthma upgrades,especially in the children,which brings extended burden and pressure to society and family.The pathogenesis and treatment of asthma are investigative hotspots constantly.Furthermore, realization of asthma in the molecule-gene level has already begun.However, many inflammatory cells and cytokines are correlated with asthma.No current treatment can reduce or prevent asthma in a long period.Further study on pathogenesy of new interference target of asthma has been generally investigated in medical science.
Objective:To observe the expression of TIMP-1(tissue inhibitor of metalloproteinase-1) in the lung tissue on the airway remodeling in the rats of the asthmatic model.Measure the airway smooth muscle area and the inner wall area standardized by the basement membrane perimeter with software of computer image analysis in the asthmatic rats model.Quantify the level of the airway remodeling with the smooth muscle area and the inner wall area of the airway.Study the effects of anti-RANTES (regulated on activation normal T cell expressed and secreted) antibody on the airway remodeling,and the new way of prevention of asthma was offered.
Method:The rats were randomly divided into three groups:asthmatic model group,anti-RANTES antibody intervention group and normal control group,each group had 12 rats.The rats of asthmatic model group and anti- RANTES antibody intervention group were immunized by intraperitoneal injection of 10% ovalbumin on the first,the seventh and the fourteenth day of the study.The rats of normal control group were given equivalent physiological saline to allergize.From the third weeks,the rats of asthmatic model group and anti-RANTES antibody intervention group were stimulated by aerosol inhalation of 1% ovalbumin (OVA),once of two days,thirty minutes every time,which were repeated six weeks.The rats of normal control group received stimulation with inhalation of physiological saline.From the fourth weeks,the rats of anti-RANTES antibody intervention group were intervened by caudal vein injection of anti-RANTES antibody,one microgramme every time before each stimulation, which were repeated five weeks.The rats of asthmatic model group and the rats of normal control group were caudal vein injection of equivalent physiological saline.To observe the behavior change of the three groups,including the action,the respiration,the appearance and the hairs of rats.The rats were anesthetized with l% pentobarbital when the end of the last stimulation.Pathological slides were prepared from left lung and stained with hematoxylin-eosin.The airway smooth muscle area (WAM),and the inner wall area (WAi) of the airway were measured and standardized by the basement membrane perimeter (Pbm). Expression of tissue inhibitor of metalloproteinase-1 were detected by immunohistochemistry.
Results:When the rats were stimulated in the asthmatic model group, they showed restlessness,scratching honor,sneezing,nose fanning,mouth breathing deepenly and quickenly,oral lip cyanochroia,and all the show above- mentioned releasing gradually after the inhalation.The HE stained section of the asthmatic model group showed mucosal edema,air duct thickening,plica to grow in number.Compared with the asthmatic model group,there were mild symptom and inflammation reaction,and decrease in air duct thicken in the anti-RANTES antibody intervention group.The rats of the normal control group showed normal diet and drinking,animation,sensitiveness,breathing stable.The airway smooth muscle area and the inner wall area of the asthmatic model group [WAm/Pbm(5.78±1.36)μm2/μm,WAi/Pbm(15.10±3.99)μm2/μm] was greater than the normal control group[WAm/Pbm(3.35±1.01)μm2/μm, WAi/Pbm(9.25±2.29)μm2/μm](all P<0.05).The airway smooth muscle area and the inner wall area of the anti-RANTES antibody intervention group [WAm/Pbm(4.12±1.21)μm2/μm,WAi/Pbm(10.01±2.69)μm2/μm]was greater than asthmatic model group (all P<0.05),but no statistical significance compared to the normal control group (all P>0.05).The relative integral optical density value of the tissue inhibitor of metalloproteinase-1 in the lung tissues was increased significantly in the asthmatic model group [1.9580±0.0927] than the normal control group [0.7623±0.1000](P<0.05).The relative integral optical density value of the tissue inhibitor of metalloproteinase-1 in the lung tissues in the anti-RANTES antibody intervention group [0.9770±0.0233] was decreased significantly than the asthmatic model group (P<0.05),but no statistical significance compared to the normal control group (P>0.05).
Conclusion:The asthmatic rats model were succeeded.The tissue inhibitor of metalloproteinase-1 had relationship with asthma airway remodeling.The anti-RANTES monoclonal antibody leads to decrease of the expression of the tissue inhibitor of metalloproteinase -1 in the lung tissue of asthma rats,lessen asthma attack,inhibit the airway inflammation,delay development of airway reconstitution.The anti-RANTES antibody takes an important role in delaying development of asthma airway remodeling.
目的:以哮喘大鼠气道模型为基础,通过研究金属蛋白酶组织抑制剂-1(TIMP-1)在气道重塑中的表达情况,并用计算机图像分析软件对气道重塑程度情况进行量化,进而探讨哮喘大鼠模型中TIMP-1与气道重塑的关系;同时应用抗正常T细胞表达和分泌的活性调节蛋白抗体(抗RANTES抗体)对哮喘大鼠进行干预,通过肺组织中TIMP-1表达的变化以及量化的气道重塑情况改变,来探讨抗RANTES抗体对气道重塑的影响,从而为哮喘的临床治疗提供动物实验依据。
方法:将36只健康雄性wistar大鼠随机分为三组:哮喘模型组、抗RANTES抗体干预组、正常对照组,每组12只。哮喘模型组和抗RANTES抗体干预组于第1、7、14天腹腔注射10%抗原液1ml(含卵白蛋白100mg、氢氧化铝100mg)进行致敏,正常对照组腹腔注射等量生理盐水致敏。从第21天起将哮喘模型组、抗RANTES抗体干预组大鼠分别置于密闭玻璃罩内,雾化吸入1%的卵白蛋白激发,每次雾化吸入30分钟,隔天1次,共6周,正常对照组用等量的生理盐水进行激发。抗RANTES抗体干预组自雾化激发后1周起即从第28天起每次激发前半小时给予尾静脉注射抗RANTES抗体1ug,哮喘模型组和正常对照组给予尾静脉注射等量生理盐水,隔天1次,共5周。观察三组实验大鼠的一般状况,包括活动、呼吸、神态、毛发等。末次激发结束后24小时用1%戊巴比妥那0.2ml/只腹腔注射麻醉各组大鼠,打开胸腔,取出左肺组织。作HE染色病理图片,光镜观察三组实验大鼠的支气管肺组织的病理改变、用计算机图像分析软件分别测定三组实验大鼠支气管基底膜周长(Pbm)、气道平滑肌面积(WAm)、气道内壁面积(WAi),并用Pbm标准化,分别以WAm/Pbm及WAi/Pbm表示气道平滑肌面积、气道内壁面积,量化气道重塑程度的改变;采用免疫组化方法测定三组实验大鼠肺组织TIMP-1的相对积分光密度的改变,观察TIMP-1的表达情况。
结果:(1)三组大鼠的一般状况:①哮喘模型组:雾化吸入卵白蛋白后大部分大鼠出现烦躁不安,搔抓颜面,鼻扇,毛发失去光泽,打喷嚏,口唇发绀,继而大鼠安静不动,弓背,张口呼吸,呼吸加深加快,前肢缩抬,四肢颤抖,口鼻流出粘液,精神不振和反应迟钝,停止雾化后上述表现逐渐缓解。②抗RANTES抗体干预组:第1周每次雾化吸入卵白蛋白后,大部分表现与哮喘模型组相似,而在应用抗RANTES抗体后,虽有烦躁、喷嚏、四肢抓挠,但无明显的呼吸困难、口唇发绀表现。③正常对照组:致敏及任意一次激发后均正常饮食、饮水,精神活泼、反应灵敏、呼吸平稳、毛色光洁,没有呼吸急促、烦躁不安或口周发绀的表现。
(2)三组大鼠气道平滑肌的比较:哮喘模型组大鼠气道平滑肌面积WAm/Pbm[(5.78±1.36)μm2/μm]明显大于正常对照组大鼠的气道平滑肌面积WAm/Pbm[(3.35±1.01)μm2/μm] (P<0.05);抗RANTES抗体干预组大鼠气道平滑肌面积WAm/Pbm[(4.12±1.21)μm2/μm]较哮喘模型组大鼠的气道平滑肌面积WAm/Pbm显著降低(P<0.05);抗RANTES抗体干预组大鼠的气道平滑肌面积WAm/Pbm与正常对照组大鼠气道平滑肌面积WAm/Pbm相比无差别(P>0.05)。
(3)三组大鼠气道内壁面积的比较:哮喘模型组大鼠气道内壁面积WAi/Pbm[(15.10±3.99)μm2/μm]明显大于正常对照组大鼠的气道气道内壁面积WAi/Pbm[(9.25±2.29)μm2/μm] (P<0.05);抗RANTES抗体干预组大鼠气道内壁面积WAi/Pbm[(10.01±2.69)μm2/μm]较哮喘模型组大鼠的气道内壁面积WAi/Pbm显著降低;抗RANTES抗体干预组大鼠的气道内壁面积WAm/Pbm与正常对照组大鼠的气道内壁面积WAi/Pbm相比无差别(P>0.05)。
(4)三组实验大鼠肺组织TIMP-1表达的变化:哮喘模型组大鼠肺组织的TIMP-1表达[积分光密度(1.9580±0.0927)]较正常对照组大鼠肺组织的TIMP-1表达[积分光密度(0.7623±0.1000)]增强(P<0.05);哮喘模型组大鼠肺组织的TIMP-1表达较抗RANTES抗体干预组大鼠肺组织的TIMP-1表达[积分光密度(0.9770±0.0233)]增强(P<0.05);抗RANTES抗体干预组大鼠肺组织的TIMP-1表达与正常对照组大鼠肺组织的TIMP-1表达相比无差别(P>0.05)。
结论:(1)哮喘大鼠模型成功;(2)哮喘模型组的大鼠肺组织中TIMP-1的表达高,可能在气道重塑中起到了重要的作用;(3)哮喘大鼠在应用抗RANTES抗体后哮喘症状缓解,气道重塑程度减轻,通过体外干预研究证实抗RANTES抗体可抑制气道重塑,它可能延缓气道重塑的发生。
Background:Asthma is an allergic disease,one common disease in chronic illness of respiratory system.Its airway is characteristed by hyperreactivity,chronic inflammation and remodeling.With the development of society,the incidence of asthma upgrades,especially in the children,which brings extended burden and pressure to society and family.The pathogenesis and treatment of asthma are investigative hotspots constantly.Furthermore, realization of asthma in the molecule-gene level has already begun.However, many inflammatory cells and cytokines are correlated with asthma.No current treatment can reduce or prevent asthma in a long period.Further study on pathogenesy of new interference target of asthma has been generally investigated in medical science.
Objective:To observe the expression of TIMP-1(tissue inhibitor of metalloproteinase-1) in the lung tissue on the airway remodeling in the rats of the asthmatic model.Measure the airway smooth muscle area and the inner wall area standardized by the basement membrane perimeter with software of computer image analysis in the asthmatic rats model.Quantify the level of the airway remodeling with the smooth muscle area and the inner wall area of the airway.Study the effects of anti-RANTES (regulated on activation normal T cell expressed and secreted) antibody on the airway remodeling,and the new way of prevention of asthma was offered.
Method:The rats were randomly divided into three groups:asthmatic model group,anti-RANTES antibody intervention group and normal control group,each group had 12 rats.The rats of asthmatic model group and anti- RANTES antibody intervention group were immunized by intraperitoneal injection of 10% ovalbumin on the first,the seventh and the fourteenth day of the study.The rats of normal control group were given equivalent physiological saline to allergize.From the third weeks,the rats of asthmatic model group and anti-RANTES antibody intervention group were stimulated by aerosol inhalation of 1% ovalbumin (OVA),once of two days,thirty minutes every time,which were repeated six weeks.The rats of normal control group received stimulation with inhalation of physiological saline.From the fourth weeks,the rats of anti-RANTES antibody intervention group were intervened by caudal vein injection of anti-RANTES antibody,one microgramme every time before each stimulation, which were repeated five weeks.The rats of asthmatic model group and the rats of normal control group were caudal vein injection of equivalent physiological saline.To observe the behavior change of the three groups,including the action,the respiration,the appearance and the hairs of rats.The rats were anesthetized with l% pentobarbital when the end of the last stimulation.Pathological slides were prepared from left lung and stained with hematoxylin-eosin.The airway smooth muscle area (WAM),and the inner wall area (WAi) of the airway were measured and standardized by the basement membrane perimeter (Pbm). Expression of tissue inhibitor of metalloproteinase-1 were detected by immunohistochemistry.
Results:When the rats were stimulated in the asthmatic model group, they showed restlessness,scratching honor,sneezing,nose fanning,mouth breathing deepenly and quickenly,oral lip cyanochroia,and all the show above- mentioned releasing gradually after the inhalation.The HE stained section of the asthmatic model group showed mucosal edema,air duct thickening,plica to grow in number.Compared with the asthmatic model group,there were mild symptom and inflammation reaction,and decrease in air duct thicken in the anti-RANTES antibody intervention group.The rats of the normal control group showed normal diet and drinking,animation,sensitiveness,breathing stable.The airway smooth muscle area and the inner wall area of the asthmatic model group [WAm/Pbm(5.78±1.36)μm2/μm,WAi/Pbm(15.10±3.99)μm2/μm] was greater than the normal control group[WAm/Pbm(3.35±1.01)μm2/μm, WAi/Pbm(9.25±2.29)μm2/μm](all P<0.05).The airway smooth muscle area and the inner wall area of the anti-RANTES antibody intervention group [WAm/Pbm(4.12±1.21)μm2/μm,WAi/Pbm(10.01±2.69)μm2/μm]was greater than asthmatic model group (all P<0.05),but no statistical significance compared to the normal control group (all P>0.05).The relative integral optical density value of the tissue inhibitor of metalloproteinase-1 in the lung tissues was increased significantly in the asthmatic model group [1.9580±0.0927] than the normal control group [0.7623±0.1000](P<0.05).The relative integral optical density value of the tissue inhibitor of metalloproteinase-1 in the lung tissues in the anti-RANTES antibody intervention group [0.9770±0.0233] was decreased significantly than the asthmatic model group (P<0.05),but no statistical significance compared to the normal control group (P>0.05).
Conclusion:The asthmatic rats model were succeeded.The tissue inhibitor of metalloproteinase-1 had relationship with asthma airway remodeling.The anti-RANTES monoclonal antibody leads to decrease of the expression of the tissue inhibitor of metalloproteinase -1 in the lung tissue of asthma rats,lessen asthma attack,inhibit the airway inflammation,delay development of airway reconstitution.The anti-RANTES antibody takes an important role in delaying development of asthma airway remodeling.
引文
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3章晓初,姚婉贞,何其华等.哮喘豚鼠气道重塑与气道高反应性的图像分析.中华结核和呼吸杂志,2001,24(2):87
4中华医学会儿科分会呼吸组,中华医学会《中华儿科杂志》编辑委员会,儿童支气管哮喘防治常规(试行).中华儿科杂志,2004,42(2):100- 106
5 Jeffery PK.Remodeling in asthma and chronic obstructive lung disease.Am J Respir Crit Care Med,2001,164(10):S28-S38
6 Takafuji S,Ishida A,Miyakuni Y,et al.Matrix metalloproteinase-9 release from human leukocytes.J Investig Allergol Clin Immunol,2003,13 (1):50-55
7 Matsumoto H,Niimi A,Take mura M,et al.Relationship of airway wall thickening to an imbalance between matrix metalloproteinase-9 and its inhibitor in asthma.Thorax,2005,60(4):277-281
8陈丽兰,庞玉生,王玲.基质金属蛋白酶-9及其抑制剂TIMP-1与支气管哮喘.中国医师杂志,2007,9(8):1150-1151
9 Wilson JW,Li X,Pain MC.The lack of distensibility of asthmatic airways. Am Rev Respir Dis,1993,148(3):806-809
10季蓉,何权瀛.对中性粒细胞在哮喘发病机制中的新认识.国外医学呼吸系统分册,2005,25(4):342-346
11 Simpson JL,Scott RJ,Boyle MJ,et al.Differential proteolytic enzyme activity in eosinophilia and neutrophilic asthma.Am J Respir Crit Care Med,2005,172(5):559-566
12周华斐,王昕昕,童夏生等.哮喘大鼠中性粒细胞基质金属蛋白酶-9和血清基质金属蛋白酶抑制剂-1的表达及地塞米松对其的影响.中国临床药理学与治疗学,2008,13(8):870-873
13 Saetta M,Di Stefano A,Rosina C,et al.Quantitative structural analysis of peripheral airways and arteries in sudden fatal asthma.Am Rev Respir Dis,1991,143(1):138-143
14 Boulet LP,Turcotte H,Laviolette M,et al.Airway hyperresponsiveness, inflammation,and subepithelial collagen deposition in recently diagnosed versus long-standing mild asthma.Influence of inhaled corticosteroids.Am J R espir Crit Care Med,2000,162(4):1308-1313
15 James AL,ParéPD,Hogg JC,et al.The mechanics of airway narrowing in asthma.Am Rev Respir Dis,1989,139(1):242-246
16 Macklem PT.Theoretial basis of airway instability.Roger S Mitchell Lecture.Chest,1995,107(3Suppl):87-88
17 Kuwano K,Bosken CH,ParéPD,et al.Small airways dimensions in asthma and in chronic obstructive pulmonary disease.Am Rev Respir Dis,1993, 148(5):1220-1225
18解卫平,丁建花,王虹等.新型KATP开放剂iptakalim对豚鼠哮喘模型气道反应性及其结构的影响.中国临床药理学与治疗学,2005,10(2):128-132
19 Djukanovic R,Poche WR,Wilson JW,et al.Mucosal inflammation in asthma.Am Rev Respir Dis,1990,142(2):434-457
20 Kay AB,Corrigan CJ.Asthma,eosinophils and neutrophils.Br Med Bull, 1992,48(1):51-64
21 Wasserman SI.Mast cell-mediated inflammation in asthma.Ann Allergy, 1989,63(6):546-550
22 Corrigan CJ,Kay AB.T cells and eosinophils in the pathogenesis of asthma. Immunol Today,1992,13(12):501-507
23 Justice JP,Borchers MT,Crosby JR,et al.Ablation of eosinophils leads to a reduction of allergen-induced pulmonary pathology.Am J physiol Lung Cell Mol Physiol,2003,248(1):169-178
24 Yins S,Taborda-Baratal L,Meng Q,et al.The kinetics of allergen-induced transcription of messenger RNA for monocyte chemotactic protein-3 and RANTES in the skin of human atopic subjects: relationship to eosinophil,T cell,and macrophage recruitment.J ExP Med,1995,181(6):2153-2159
25许从峰,熊思东,杨英珍等.RANTEs生物功能的多样性.生命的化学, 2001,21(5):353-354
26 Kayaba H,Chihara J.Eosinophils and related chemokines.Rinsho Byori, Japanese Journal of Clinical Pathology,2001,49(4):370-375
27 Teran LM,Noso N,Carroll M,et al.Eosinophil recruitment following allergen challenge is associated with the release of chemokine RANTES into asthmatic airways.J Immunol,1996,157(4):1806-1812
28 Meurer R,Van RiperG,Feeney W,et al.Formation of eosinophilic and monocytic intradermal inflammatory sites in the dog by injection of human RANTES but not human monocyte chemoattractant protein-1, human macrophage inflammtory protein 1 alpha,or human interleukin 8.J Exp Med,1993,178(6):1913-1921
29 Nakamura H,Weiss ST,Israel E,et al.Eotaxin and impaired lung function in asthma.Am J Respir Crit Care Med,1999,160(6):1952-1956
30 Rot A,Krieger M,Brunner T,et al.RANTES and macrophage inflammatory
31 protein-1 alpha induce the migration and activation of normal human eosinophil granulocytes.J Exp Med,1992,176(6):1489-1495
32 Schall TJ,Bacon K,Toy KJ,et al.Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES.Nature, 1990,347(6294):669-671
33 Taub DD,Conlon K,Lioyd AR,et al.Preferential migration of activated CD4+ and CD8+ T cells in response to MIP-1 alpha and MIP-1 beta. Science,1993,260(5106):355-358
34 Gonzalo JA,Lloyd CM,Wen D,et al.The coordinated action of CC chemokines in the lung orchestrates allergic inflammation and airway hyperresponsiveness.J Exp Med,1998,188(1):157-167
35李学明,黄建安.哮喘患者外周血趋化因子RANTES、MIP-l和IL-8变化的临床意义
36孔祥永,贾天明,栾斌等.气道重塑与支气管哮喘的治疗.国外医学呼吸系统分册,2005,12(5):40-42
37 Leigh R,Vethanayagam D,Yoshida M,et al.Effect of montelukast and budesonide montelukast on airway responses and airway inflammation in asthma.Am J Respir Crit care Med,2002,166(9):1212-1217
38 Lamkhioued B,Renzi PM,Abi-Younes S,et al.Increased expression of eotaxin in bronchoalveolar lavage and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation.J Immunol,1997, 159(9):4593-4601
39 Becker S,Reed W,Henderson FW,et al.RSV infection of human airway epithelial cells causes production of the beta-chemokine RANTES.Am J Physiol,1997,272(3):512-520
1 KitayamaJ, Mackay CR,Ponath PD,et al.The C-C chemokine receptor CCR3 participates in stimulation of eosinophil arrest on inflammatory endothelium in shear flow.Clin Invest,1998,101(9):2017-2024
2 Schall TJ,Jongstra J,Dyer BJ,et al.A human T cell-specific molecule is a member of a new gene family.J Immunol,1988,141(3):1018-1025
3刘疗,杨云霞.调控RANTES的信号转导途径的研究进展.西部医学,2009,21(6):1031-1033
4 Ward SG,Westwick J.Chemokines:understanding their role in T-lymphocyte biology.Biochem J,1998,333(3):457-470
5姚玲,龚卫娟.RANTES拮抗剂的研究进展.国际免疫学杂志,2006,29 (3):149-152
6 Matsukura S,Stellato C,Plitt JR,et al.Activation of eotaxin gene transcription by NF-kappa B and STAT6 in human airway epithelial cells. J Immunol,1999,163(12):6876-6883
7 Elsner J, Kapp A.Modulation of eosinophil effector functions: the potential role of monoclonal antibodies and chemokine receptor antagonists.Allergy Asthma Proc,2000,21(6):345-349
8 Ali S,Kaur J,Patel KD.Intercellular cell adhesion molecule-1,vascular cell adhesion molecule-1,and regulated on activation normal T cell expressedand secreted are expressed by human breast carcinoma cells and support eosinophil adhesion and activation.Am J Pathol,2000,157(1):313-321
9 Logan MR,Lacy P,Bablitz B,et a1.Expression of eosinophil target SNAREs as potential cognate receptors for vesicle-associated membrane protein-2 in exocytosis.J Allergy Clin Immunol,2002,109(2):299-306
10 Gunter Wolf,Fuad N,Ziyadeh,et al.Angiotensin II stimulates expression of the chemokine RANTES in rat glomerular endothelial cells.Role of the angiotensin type 2 receptor.J Clin Invest,1997,100(5):1047-1058
11 Panzer U,Schneider A,Wilken J,et a1.The chemokine receptor antagonist AOP-RANTES reduces monocyte infiltration in experimental glomerulonephritis.Kidney Int,1999,56(6):2107-2115
12 McGovern V.Taking a world view of asthma.Environ Health Perspect, 2002,110(9):514-515
13 Géninp P,AlgartéM,Roof P,et a1.Regulation of RANTES chemokine gene expression requires cooperativity between NF-kappaB and IFN- regulatory factor transcription factors.J Immunol,2000,164(10):5352-5361
14杨仕贵.趋化因子RANTES及其受体与哮喘.国外医学免疫学分册. 2003,6(3):126-129
15 Bacon KB,Premack BA,Gargner P,et a1.Activation of dual T cell signaling pathways by the chemokine RANTES.J Science,1995,269(5231):1727- 1730
16 Kunkel SL,Ajuebor MN,Hogaboam CM,et al.The chemokine RANTES is a crucial mediator of the progression from acute to chronic colitis in the rat. J Immunol,2001,166(1):552-558
17 Beck LA,Dalke S,Leiferman K,et al.Cutaneous injection of RANTES causes eosinophil recruitment:comparison of nonallergic and allergic human subjects.J Immunol,1997,159(6):2962-2972
18刘传合,陈虹,胡良平等.趋化因子单核苷酸多态性与中国儿童哮喘的相关性研究,中华儿科杂志,2005,43(6):462-463
19 Barczyk A,Sozańska E,Farnik M,et al.Increased levels of RANTES in induced sputum of chronic asthma but not in seasonal grass pollen-inducedrhinitis.Pneumonol Alergol Pol,2006,74(2):191-196
20 Rojas-Ramos E,Avalos AF,Perez-Fernandez L,et al.Role of the chemokines RANTES,monocyte chemotacticproteins-3 and -4,and eotaxins-1 and -2 in childhood asthma.Eur Respir J,2003,22(2):310-316
21 Simson L,Foster PS.Chemokine and cytokine cooperativity:eosinophil migration in the asthmatic response.Immunol Cell Biol,2000,78(4):415- 422
22 Alam R,York J,Boyars M,et al.Increased MCP-1,RANTES,and MIP-1 alpha in bronchoalveolar lavage fluid of allergic asthmatic patients.Am J Respir Crit Care Med,1996,153(4):1398-1404
23 Fryer AA,Spiteri MA,Bianco A,et al.The 403G/A promoter polymorphism in the RANTES gene is associated with atopy and asthma.Genes Immun, 2000,1(8):509-514
24 Conti P,DiGioacchino M.MCP-1 and RANTES are mediators of acute and chronic inflammation.Allergy Asthma Proc,2001,22(3):133-137
25 The Collaborative Study on the Genetics of Asthma (CSGA).A genome- wide search for asthma susceptibility loci in ethnically diverse population. Nat Genet,1997,15(4):389-392
26 Miyamasu M,Sekiya T,Ohta K,et al.Variations in the human CC chemokine eotaxin gene.Genes Immun,2001,2(8):461-463
27文惯宇,赵德育,田曼.RANTES启动子-403G/A基因多态性与呼吸道合胞病毒致毛细支气管炎的相关性研究.南京医科大学学报(自然科学版),2007,27(5):487-490
28 Hyvarinen M,Piippo-Savolainen E,Korhonen K,et al.Teenage asthma after severe infantile bronchiolitis or pneumonia.Acta Paediatr,2005,94(10): 1378-1383
29 Szalai C,Kozma GT,Nagy A,et al.Polymorphism in the gene regulatory region of MCP-1 is associated with asthma susceptibility and severity. Allergy Clin Immunol,2001,108(3):375-381
30 Ying S,Meng Q,Zeibecoglou K,et al.Eosinophil chemotactic chemokines (eotaxin,eotaxin-2,RANTES,monocyte chemoattractant protei-3(MCP-3),and MCP-4),and C-C chemokine receptor 3 expression in bronchial biopsies from atopic and nonatopic (Intrinsic ) asthmatics.J Immunol, 1999,163(11):6321-6329
31 Bello I,Rizo MR,Badel CB,et al.In vitro interferon gamma regulation of CCR-3 mRNA expression in peripheral blood leukocytes from atopic asthmatics.Clin Exp Med,2001,1(2):75-80
32 Penido C,Castro-Faria-Neto HC,Vieira-de-Abreu A,et al.LPS induces eosinophil migration via CCR3 signaling through a mechanism independent of RANTES and Eotaxin.Am J Respir Cell Mol Biol,2001,25 (6):707-716
33 Schuh JM,Blease K,Hogaboam CM.The role of CC chemokine receptor 5(CCR5) and RANTES /CCL5 during chronic fungal asthma in mice. FASEB J,2002,16(2):228-230
34 Bertrand CP.Asthma:where beyond steroids?.Curr Opin Chem Biol,2000,4 (4):407-411
35 Menzies-Gow A,Robinson DS.Eosinophils,eosinophilic cytokines (interleukin-5),and antieosinophilic therapy in asthma .Curr Opin Pulm Med,2002,8(1):33-38
36 Tekkanat KK,Maassab H,Miller A,et al.RANTES(CCL5) production during primary respiratory syncytial virus infection exacerbates airway disease.Eur J Immunol,2002,32(11):3276-3284
37孙志成,郑红,李世荣等.Met-RANTES对内皮细胞与T细胞相互作用的影响.中华免疫学杂志,2005,21(11):846-849
2王丽新,吴银根.实验性哮喘大鼠气道重塑的形态学变化.中西医结合学报,2003,1(1):62-65
3章晓初,姚婉贞,何其华等.哮喘豚鼠气道重塑与气道高反应性的图像分析.中华结核和呼吸杂志,2001,24(2):87
4中华医学会儿科分会呼吸组,中华医学会《中华儿科杂志》编辑委员会,儿童支气管哮喘防治常规(试行).中华儿科杂志,2004,42(2):100- 106
5 Jeffery PK.Remodeling in asthma and chronic obstructive lung disease.Am J Respir Crit Care Med,2001,164(10):S28-S38
6 Takafuji S,Ishida A,Miyakuni Y,et al.Matrix metalloproteinase-9 release from human leukocytes.J Investig Allergol Clin Immunol,2003,13 (1):50-55
7 Matsumoto H,Niimi A,Take mura M,et al.Relationship of airway wall thickening to an imbalance between matrix metalloproteinase-9 and its inhibitor in asthma.Thorax,2005,60(4):277-281
8陈丽兰,庞玉生,王玲.基质金属蛋白酶-9及其抑制剂TIMP-1与支气管哮喘.中国医师杂志,2007,9(8):1150-1151
9 Wilson JW,Li X,Pain MC.The lack of distensibility of asthmatic airways. Am Rev Respir Dis,1993,148(3):806-809
10季蓉,何权瀛.对中性粒细胞在哮喘发病机制中的新认识.国外医学呼吸系统分册,2005,25(4):342-346
11 Simpson JL,Scott RJ,Boyle MJ,et al.Differential proteolytic enzyme activity in eosinophilia and neutrophilic asthma.Am J Respir Crit Care Med,2005,172(5):559-566
12周华斐,王昕昕,童夏生等.哮喘大鼠中性粒细胞基质金属蛋白酶-9和血清基质金属蛋白酶抑制剂-1的表达及地塞米松对其的影响.中国临床药理学与治疗学,2008,13(8):870-873
13 Saetta M,Di Stefano A,Rosina C,et al.Quantitative structural analysis of peripheral airways and arteries in sudden fatal asthma.Am Rev Respir Dis,1991,143(1):138-143
14 Boulet LP,Turcotte H,Laviolette M,et al.Airway hyperresponsiveness, inflammation,and subepithelial collagen deposition in recently diagnosed versus long-standing mild asthma.Influence of inhaled corticosteroids.Am J R espir Crit Care Med,2000,162(4):1308-1313
15 James AL,ParéPD,Hogg JC,et al.The mechanics of airway narrowing in asthma.Am Rev Respir Dis,1989,139(1):242-246
16 Macklem PT.Theoretial basis of airway instability.Roger S Mitchell Lecture.Chest,1995,107(3Suppl):87-88
17 Kuwano K,Bosken CH,ParéPD,et al.Small airways dimensions in asthma and in chronic obstructive pulmonary disease.Am Rev Respir Dis,1993, 148(5):1220-1225
18解卫平,丁建花,王虹等.新型KATP开放剂iptakalim对豚鼠哮喘模型气道反应性及其结构的影响.中国临床药理学与治疗学,2005,10(2):128-132
19 Djukanovic R,Poche WR,Wilson JW,et al.Mucosal inflammation in asthma.Am Rev Respir Dis,1990,142(2):434-457
20 Kay AB,Corrigan CJ.Asthma,eosinophils and neutrophils.Br Med Bull, 1992,48(1):51-64
21 Wasserman SI.Mast cell-mediated inflammation in asthma.Ann Allergy, 1989,63(6):546-550
22 Corrigan CJ,Kay AB.T cells and eosinophils in the pathogenesis of asthma. Immunol Today,1992,13(12):501-507
23 Justice JP,Borchers MT,Crosby JR,et al.Ablation of eosinophils leads to a reduction of allergen-induced pulmonary pathology.Am J physiol Lung Cell Mol Physiol,2003,248(1):169-178
24 Yins S,Taborda-Baratal L,Meng Q,et al.The kinetics of allergen-induced transcription of messenger RNA for monocyte chemotactic protein-3 and RANTES in the skin of human atopic subjects: relationship to eosinophil,T cell,and macrophage recruitment.J ExP Med,1995,181(6):2153-2159
25许从峰,熊思东,杨英珍等.RANTEs生物功能的多样性.生命的化学, 2001,21(5):353-354
26 Kayaba H,Chihara J.Eosinophils and related chemokines.Rinsho Byori, Japanese Journal of Clinical Pathology,2001,49(4):370-375
27 Teran LM,Noso N,Carroll M,et al.Eosinophil recruitment following allergen challenge is associated with the release of chemokine RANTES into asthmatic airways.J Immunol,1996,157(4):1806-1812
28 Meurer R,Van RiperG,Feeney W,et al.Formation of eosinophilic and monocytic intradermal inflammatory sites in the dog by injection of human RANTES but not human monocyte chemoattractant protein-1, human macrophage inflammtory protein 1 alpha,or human interleukin 8.J Exp Med,1993,178(6):1913-1921
29 Nakamura H,Weiss ST,Israel E,et al.Eotaxin and impaired lung function in asthma.Am J Respir Crit Care Med,1999,160(6):1952-1956
30 Rot A,Krieger M,Brunner T,et al.RANTES and macrophage inflammatory
31 protein-1 alpha induce the migration and activation of normal human eosinophil granulocytes.J Exp Med,1992,176(6):1489-1495
32 Schall TJ,Bacon K,Toy KJ,et al.Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES.Nature, 1990,347(6294):669-671
33 Taub DD,Conlon K,Lioyd AR,et al.Preferential migration of activated CD4+ and CD8+ T cells in response to MIP-1 alpha and MIP-1 beta. Science,1993,260(5106):355-358
34 Gonzalo JA,Lloyd CM,Wen D,et al.The coordinated action of CC chemokines in the lung orchestrates allergic inflammation and airway hyperresponsiveness.J Exp Med,1998,188(1):157-167
35李学明,黄建安.哮喘患者外周血趋化因子RANTES、MIP-l和IL-8变化的临床意义
36孔祥永,贾天明,栾斌等.气道重塑与支气管哮喘的治疗.国外医学呼吸系统分册,2005,12(5):40-42
37 Leigh R,Vethanayagam D,Yoshida M,et al.Effect of montelukast and budesonide montelukast on airway responses and airway inflammation in asthma.Am J Respir Crit care Med,2002,166(9):1212-1217
38 Lamkhioued B,Renzi PM,Abi-Younes S,et al.Increased expression of eotaxin in bronchoalveolar lavage and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation.J Immunol,1997, 159(9):4593-4601
39 Becker S,Reed W,Henderson FW,et al.RSV infection of human airway epithelial cells causes production of the beta-chemokine RANTES.Am J Physiol,1997,272(3):512-520
1 KitayamaJ, Mackay CR,Ponath PD,et al.The C-C chemokine receptor CCR3 participates in stimulation of eosinophil arrest on inflammatory endothelium in shear flow.Clin Invest,1998,101(9):2017-2024
2 Schall TJ,Jongstra J,Dyer BJ,et al.A human T cell-specific molecule is a member of a new gene family.J Immunol,1988,141(3):1018-1025
3刘疗,杨云霞.调控RANTES的信号转导途径的研究进展.西部医学,2009,21(6):1031-1033
4 Ward SG,Westwick J.Chemokines:understanding their role in T-lymphocyte biology.Biochem J,1998,333(3):457-470
5姚玲,龚卫娟.RANTES拮抗剂的研究进展.国际免疫学杂志,2006,29 (3):149-152
6 Matsukura S,Stellato C,Plitt JR,et al.Activation of eotaxin gene transcription by NF-kappa B and STAT6 in human airway epithelial cells. J Immunol,1999,163(12):6876-6883
7 Elsner J, Kapp A.Modulation of eosinophil effector functions: the potential role of monoclonal antibodies and chemokine receptor antagonists.Allergy Asthma Proc,2000,21(6):345-349
8 Ali S,Kaur J,Patel KD.Intercellular cell adhesion molecule-1,vascular cell adhesion molecule-1,and regulated on activation normal T cell expressedand secreted are expressed by human breast carcinoma cells and support eosinophil adhesion and activation.Am J Pathol,2000,157(1):313-321
9 Logan MR,Lacy P,Bablitz B,et a1.Expression of eosinophil target SNAREs as potential cognate receptors for vesicle-associated membrane protein-2 in exocytosis.J Allergy Clin Immunol,2002,109(2):299-306
10 Gunter Wolf,Fuad N,Ziyadeh,et al.Angiotensin II stimulates expression of the chemokine RANTES in rat glomerular endothelial cells.Role of the angiotensin type 2 receptor.J Clin Invest,1997,100(5):1047-1058
11 Panzer U,Schneider A,Wilken J,et a1.The chemokine receptor antagonist AOP-RANTES reduces monocyte infiltration in experimental glomerulonephritis.Kidney Int,1999,56(6):2107-2115
12 McGovern V.Taking a world view of asthma.Environ Health Perspect, 2002,110(9):514-515
13 Géninp P,AlgartéM,Roof P,et a1.Regulation of RANTES chemokine gene expression requires cooperativity between NF-kappaB and IFN- regulatory factor transcription factors.J Immunol,2000,164(10):5352-5361
14杨仕贵.趋化因子RANTES及其受体与哮喘.国外医学免疫学分册. 2003,6(3):126-129
15 Bacon KB,Premack BA,Gargner P,et a1.Activation of dual T cell signaling pathways by the chemokine RANTES.J Science,1995,269(5231):1727- 1730
16 Kunkel SL,Ajuebor MN,Hogaboam CM,et al.The chemokine RANTES is a crucial mediator of the progression from acute to chronic colitis in the rat. J Immunol,2001,166(1):552-558
17 Beck LA,Dalke S,Leiferman K,et al.Cutaneous injection of RANTES causes eosinophil recruitment:comparison of nonallergic and allergic human subjects.J Immunol,1997,159(6):2962-2972
18刘传合,陈虹,胡良平等.趋化因子单核苷酸多态性与中国儿童哮喘的相关性研究,中华儿科杂志,2005,43(6):462-463
19 Barczyk A,Sozańska E,Farnik M,et al.Increased levels of RANTES in induced sputum of chronic asthma but not in seasonal grass pollen-inducedrhinitis.Pneumonol Alergol Pol,2006,74(2):191-196
20 Rojas-Ramos E,Avalos AF,Perez-Fernandez L,et al.Role of the chemokines RANTES,monocyte chemotacticproteins-3 and -4,and eotaxins-1 and -2 in childhood asthma.Eur Respir J,2003,22(2):310-316
21 Simson L,Foster PS.Chemokine and cytokine cooperativity:eosinophil migration in the asthmatic response.Immunol Cell Biol,2000,78(4):415- 422
22 Alam R,York J,Boyars M,et al.Increased MCP-1,RANTES,and MIP-1 alpha in bronchoalveolar lavage fluid of allergic asthmatic patients.Am J Respir Crit Care Med,1996,153(4):1398-1404
23 Fryer AA,Spiteri MA,Bianco A,et al.The 403G/A promoter polymorphism in the RANTES gene is associated with atopy and asthma.Genes Immun, 2000,1(8):509-514
24 Conti P,DiGioacchino M.MCP-1 and RANTES are mediators of acute and chronic inflammation.Allergy Asthma Proc,2001,22(3):133-137
25 The Collaborative Study on the Genetics of Asthma (CSGA).A genome- wide search for asthma susceptibility loci in ethnically diverse population. Nat Genet,1997,15(4):389-392
26 Miyamasu M,Sekiya T,Ohta K,et al.Variations in the human CC chemokine eotaxin gene.Genes Immun,2001,2(8):461-463
27文惯宇,赵德育,田曼.RANTES启动子-403G/A基因多态性与呼吸道合胞病毒致毛细支气管炎的相关性研究.南京医科大学学报(自然科学版),2007,27(5):487-490
28 Hyvarinen M,Piippo-Savolainen E,Korhonen K,et al.Teenage asthma after severe infantile bronchiolitis or pneumonia.Acta Paediatr,2005,94(10): 1378-1383
29 Szalai C,Kozma GT,Nagy A,et al.Polymorphism in the gene regulatory region of MCP-1 is associated with asthma susceptibility and severity. Allergy Clin Immunol,2001,108(3):375-381
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