中华儿科杂志
中華兒科雜誌
중화인과잡지
Chinese Journal of Pediatrics
2015年
6期
453-458
,共6页
钱娟%李本尚%殷敏智%沈萍%孙锟
錢娟%李本尚%慇敏智%瀋萍%孫錕
전연%리본상%은민지%침평%손곤
儿童%室间隔缺损%尿激酶型纤溶酶原激活剂%纤溶酶原激活物抑制物1
兒童%室間隔缺損%尿激酶型纖溶酶原激活劑%纖溶酶原激活物抑製物1
인동%실간격결손%뇨격매형섬용매원격활제%섬용매원격활물억제물1
Child%Heart septal defects,ventricular%Urokinase-type plasminogen activator%Plasminogen activator inhibitor 1
目的 分析尿激酶型纤溶酶原激活物(uPA)及纤溶酶原激活物抑制物1(PAI-1)在正常三尖瓣组织和假性室隔瘤(VSA)中的表达,初步探索VSA形成的规律.方法 2008年1月至2010年6月在上海仁济医院和上海儿童医学中心手术分别获得正常人心脏三尖瓣隔瓣7例和膜周型室间隔缺损伴VSA组织33例.免疫病理方法检测uPA和PAI-1的分布和表达,Western blot检测uPA和PAI-1的蛋白合成;荧光定量PCR方法检测uPA和PAI-1 mRNA的表达.组间比较采用t检验.结果 VSA组织具有完整的内皮结构,在内皮下层出现多层相间的肉芽样组织(致密层和疏松层),uPA主要分布在正常三尖瓣的纤维层和VSA组织致密层,部分血管平滑肌层中亦有表达,PAI-1广泛表达在正常三尖瓣纤维层和海绵层,VSA致密层和稀疏层.9例uPA高表达组uPA表达率与正常三尖瓣组比较,差异有统计学意义[(74.6±11.8)％比(49.5±7.4)％;t=3.87,P=0.003];6例uPA低表达组uPA表达率与正常三尖瓣组比较,差异有统计学意义[(10.3±3.1)％比(49.5±7.4)％;t=11.78,P=0.000].PAI-1表达率在uPA低表达组和正常三尖瓣组间差异有统计学意义[(55.2±1.7)％比(50.8±3.8)％;t=2.55,P=0.034],在uPA高表达组和正常三尖瓣组差异无统计学意义[(47.8±3.6)％比(50.8±3.8)％;t=1.366,P=0.199].Western blot结果显示,uPA高表达组(n=5),uPA表达灰度值和uPA/PAI-1比值分别为94±16,4.26±2.04,与正常三尖瓣组比较,两者差异均有统计学意义(t=2.669、3.670,P均＜0.05);uPA低表达组(n=6)uPA/PAI-1比值与正常三尖瓣比较,两者差异有统计学意义(t =6.309,P＜0.01).2例VSA(uPA高表达组)组织PAI-1相对拷贝数和uPA/PAI-1相对拷贝数比值分别为7.43±0.72,0.45±0.04,与正常瓣膜比较,差异均有明显统计学意义(t=6.499、13.269,P均＜0.01);另5例VSA(uPA低表达组)组织uPA/PAI-1相对拷贝数比值为4.38±1.41,与正常瓣膜比较,差异有统计学意义(t=3.494,P＜0.05).结论 uPA及抑制物系统在VSA形成过程中起重要作用,参与瘤体的形成和纤维增殖过程.
目的 分析尿激酶型纖溶酶原激活物(uPA)及纖溶酶原激活物抑製物1(PAI-1)在正常三尖瓣組織和假性室隔瘤(VSA)中的錶達,初步探索VSA形成的規律.方法 2008年1月至2010年6月在上海仁濟醫院和上海兒童醫學中心手術分彆穫得正常人心髒三尖瓣隔瓣7例和膜週型室間隔缺損伴VSA組織33例.免疫病理方法檢測uPA和PAI-1的分佈和錶達,Western blot檢測uPA和PAI-1的蛋白閤成;熒光定量PCR方法檢測uPA和PAI-1 mRNA的錶達.組間比較採用t檢驗.結果 VSA組織具有完整的內皮結構,在內皮下層齣現多層相間的肉芽樣組織(緻密層和疏鬆層),uPA主要分佈在正常三尖瓣的纖維層和VSA組織緻密層,部分血管平滑肌層中亦有錶達,PAI-1廣汎錶達在正常三尖瓣纖維層和海綿層,VSA緻密層和稀疏層.9例uPA高錶達組uPA錶達率與正常三尖瓣組比較,差異有統計學意義[(74.6±11.8)％比(49.5±7.4)％;t=3.87,P=0.003];6例uPA低錶達組uPA錶達率與正常三尖瓣組比較,差異有統計學意義[(10.3±3.1)％比(49.5±7.4)％;t=11.78,P=0.000].PAI-1錶達率在uPA低錶達組和正常三尖瓣組間差異有統計學意義[(55.2±1.7)％比(50.8±3.8)％;t=2.55,P=0.034],在uPA高錶達組和正常三尖瓣組差異無統計學意義[(47.8±3.6)％比(50.8±3.8)％;t=1.366,P=0.199].Western blot結果顯示,uPA高錶達組(n=5),uPA錶達灰度值和uPA/PAI-1比值分彆為94±16,4.26±2.04,與正常三尖瓣組比較,兩者差異均有統計學意義(t=2.669、3.670,P均＜0.05);uPA低錶達組(n=6)uPA/PAI-1比值與正常三尖瓣比較,兩者差異有統計學意義(t =6.309,P＜0.01).2例VSA(uPA高錶達組)組織PAI-1相對拷貝數和uPA/PAI-1相對拷貝數比值分彆為7.43±0.72,0.45±0.04,與正常瓣膜比較,差異均有明顯統計學意義(t=6.499、13.269,P均＜0.01);另5例VSA(uPA低錶達組)組織uPA/PAI-1相對拷貝數比值為4.38±1.41,與正常瓣膜比較,差異有統計學意義(t=3.494,P＜0.05).結論 uPA及抑製物繫統在VSA形成過程中起重要作用,參與瘤體的形成和纖維增殖過程.
목적 분석뇨격매형섬용매원격활물(uPA)급섬용매원격활물억제물1(PAI-1)재정상삼첨판조직화가성실격류(VSA)중적표체,초보탐색VSA형성적규률.방법 2008년1월지2010년6월재상해인제의원화상해인동의학중심수술분별획득정상인심장삼첨판격판7례화막주형실간격결손반VSA조직33례.면역병리방법검측uPA화PAI-1적분포화표체,Western blot검측uPA화PAI-1적단백합성;형광정량PCR방법검측uPA화PAI-1 mRNA적표체.조간비교채용t검험.결과 VSA조직구유완정적내피결구,재내피하층출현다층상간적육아양조직(치밀층화소송층),uPA주요분포재정상삼첨판적섬유층화VSA조직치밀층,부분혈관평활기층중역유표체,PAI-1엄범표체재정상삼첨판섬유층화해면층,VSA치밀층화희소층.9례uPA고표체조uPA표체솔여정상삼첨판조비교,차이유통계학의의[(74.6±11.8)％비(49.5±7.4)％;t=3.87,P=0.003];6례uPA저표체조uPA표체솔여정상삼첨판조비교,차이유통계학의의[(10.3±3.1)％비(49.5±7.4)％;t=11.78,P=0.000].PAI-1표체솔재uPA저표체조화정상삼첨판조간차이유통계학의의[(55.2±1.7)％비(50.8±3.8)％;t=2.55,P=0.034],재uPA고표체조화정상삼첨판조차이무통계학의의[(47.8±3.6)％비(50.8±3.8)％;t=1.366,P=0.199].Western blot결과현시,uPA고표체조(n=5),uPA표체회도치화uPA/PAI-1비치분별위94±16,4.26±2.04,여정상삼첨판조비교,량자차이균유통계학의의(t=2.669、3.670,P균＜0.05);uPA저표체조(n=6)uPA/PAI-1비치여정상삼첨판비교,량자차이유통계학의의(t =6.309,P＜0.01).2례VSA(uPA고표체조)조직PAI-1상대고패수화uPA/PAI-1상대고패수비치분별위7.43±0.72,0.45±0.04,여정상판막비교,차이균유명현통계학의의(t=6.499、13.269,P균＜0.01);령5례VSA(uPA저표체조)조직uPA/PAI-1상대고패수비치위4.38±1.41,여정상판막비교,차이유통계학의의(t=3.494,P＜0.05).결론 uPA급억제물계통재VSA형성과정중기중요작용,삼여류체적형성화섬유증식과정.
Objective The exact mechanisms of defect closure in patients with perimembranous ventricular septal defect (PMVSD) remain unknown.We hypothesized that the expression of urokinase type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) may mediate extracellular matrix (ECM) remodeling in aneurysms.Method Seven normal heart tricuspid septal leaflet and 33 aneurysms were collected in Shanghai Renji Hospital and Shanghai Children's Medical Center from January 2008 to June 2010.Immunohistochemical expression of uPA and PAI-1 in 4 normal heart valvular tissues and 15 aneurysms was detected with immunohistochemical methods.The expression of uPA and PAI-1 mRNA in 3 normal heart valvular tissues and 7 aneurysms was studied by real time fluorescent PCR;the protein expression of uPA and PAI-1 in 4 normal heart valvular tissues and 11 aneurysms was tested with Western blotting.Result The surface of the aneurysms were completely covered by endothelial cells.Two types of granulation tissue,myxoid and fibrous,were associated with the aneurismal formation.uPA were recognized predominantly in valvar interstitial cells (VICs) which located mainly in regions adjacent to the endothelium and smooth muscle cells of blood vessels.PAI-1 was found in both VICs which located mainly in granulation tissue and endothelial cells.Nine aneurysms expressed a higher uPA activity than 4 normal valvular tissues ((74.6 ± 11.8) ％ vs.(49.5 ± 7.4) ％;t =3.87,P =0.003) and six aneurysms expressed a low uPA activity ((10.3±3.1)％ vs.(49.5±7.4)％;t=11.78,P=0.000) andahighPAI-1 activity ((55.2±1.7) ％ vs.(50.8 ± 3.8) ％;t =2.55,P =0.034) using immunohistochemical methods.uPA / PAI-1 ratio of protein expression tested by Western blot was 0.88 ± 0.22 in four normal heart vavular tissues;five aneurysms expressed high uPA activity and low PAI-1 activity and uPA/PAI-1 ratio was 4.26 ± 2.04;while the other 6 cases expressed low uPA activity and high PAI-1 activity and uPA/PAI-1 ratio was 0.30 ± 0.07;the difference among the three groups was statistically significant (P ＜ 0.05).The rate of uPA/PAI-I in relative copy of mRNA expression among normal heart valvular tissue,high uPA expressed aneurysms and low uPA expressed aneurysms are also significantly different (2.14 ± 0.17 vs.0.45 ± 0.04;2.14 ± 0.17vs.4.38 ± 1.41,P ＜ 0.05) respectively.Conclusion The expression of uPA and PAI-1 in VICs suggests that interactions among these molecules contribute to the aneurysm formation and development.This provides a potential mechanism for defect closure in patients with PMVSD.
