CAJ | 학술논문

目的观察CD40、CD40L和基质金属蛋白酶9(MMP9)在人类颈动脉粥样硬化斑块中的表达情况,探讨CD40、CD40L在斑块稳定性中的作用及与MMP9表达的关系.方法将因颈动脉粥样硬化狭窄(>70%)行颈动脉内膜切除术的37例手术标本分为有临床脑卒中事件组(A组,n=20)和无临床脑卒中事件组(B组,n=17),另设尸检正常颈动脉(C组,n=11)作为对照组,采用实时荧光定量聚合酶链反应法检测各组斑块中CD40、CD40L和MMP9 mRNA水平,Western blot检测各组CD40、CD40L和MMP9蛋白表达水平,免疫组织化学检测各组CD40、CD40L和MMP9表达及分布,并对CD40、CD40L与MMP9的转录及表达水平作相关性分析.结果 A、B、C 3组的CD40mRNA的相对表达量分别为:2.41±0.43、1.03±0.38和0.31±0.12;MMP9 mRNA的相对表达量分别为:6.88±1.57、1.90±0.44和0.39±0.12.A组CD40、MMP9的mRNA水平高于B组(FCD40=105.183,FMMP9=160.395,P=0.000),B组高于C组(FCD40=37.307,FMMP9=124.093,P=0.000);CD40与MMP9的mRNA水平有正的直线相关关系(r=0.929,P=0.000),但各组CD40L的mRNA水平差异无统计学意义(FA-B=0.033,P=0.857;FB-C=1.564,P=0.767;FA-C=0.219,P=0.644).A组CD40、CD40L和MMP9的蛋白表达高于B组(FCD40=104.100,P=0.000;FCD40L=129.932,P=0.000;FMMP9=13.565,P=0.021),B组高于C组(FCD40=115.848,P=0.000;FCD40L=30.482,P=0.005;FMMP9=35.557,P=0.004).免疫组织化学示CD40、CD40L和MMP9的阳性表达面积A组大于B组(FCD40=39.451,FCD40L=57.606,FMMP9=30.711,P=0.000),B组大于C组(FCD40=95.066,FCD40L=59.081,FMMP9=145.758,P=0.000);CD40、CD40L与MMP9的阳性表达面积有正相关关系(rCD40L-CD40L=0.963,rCD40-MMP9=0.959,rCD40L-MMP9=0.929,P=0.000),并且CD40、CD40L和MMP9均在斑块肩部表达明显增多.结论 CD40-CD40L在动脉粥样硬化的形成和斑块稳定性中起重要作用;其可能通过上调MMP9导致斑块不稳定;CD40L可能是通过转录后修饰影响CD40L的表达,从而产生生物学效应.
목적 관찰CD40、CD40L화기질금속단백매9(MMP9)재인류경동맥죽양경화반괴중적표체정황,탐토CD40、CD40L재반괴은정성중적작용급여MMP9표체적관계.방법 장인경동맥죽양경화협착(>70%)행경동맥내막절제술적37례수술표본분위유림상뇌졸중사건조(A조,n=20)화무림상뇌졸중사건조(B조,n=17),령설시검정상경동맥(C조,n=11)작위대조조,채용실시형광정량취합매련반응법검측각조반괴중CD40、CD40L화MMP9 mRNA수평,Western blot검측각조CD40、CD40L화MMP9단백표체수평,면역조직화학검측각조CD40、CD40L화MMP9표체급분포,병대CD40、CD40L여MMP9적전록급표체수평작상관성분석.결과 A、B、C 3조적CD40mRNA적상대표체량분별위:2.41±0.43、1.03±0.38화0.31±0.12;MMP9 mRNA적상대표체량분별위:6.88±1.57、1.90±0.44화0.39±0.12.A조CD40、MMP9적mRNA수평고우B조(FCD40=105.183,FMMP9=160.395,P=0.000),B조고우C조(FCD40=37.307,FMMP9=124.093,P=0.000);CD40여MMP9적mRNA수평유정적직선상관관계(r=0.929,P=0.000),단각조CD40L적mRNA수평차이무통계학의의(FA-B=0.033,P=0.857;FB-C=1.564,P=0.767;FA-C=0.219,P=0.644).A조CD40、CD40L화MMP9적단백표체고우B조(FCD40=104.100,P=0.000;FCD40L=129.932,P=0.000;FMMP9=13.565,P=0.021),B조고우C조(FCD40=115.848,P=0.000;FCD40L=30.482,P=0.005;FMMP9=35.557,P=0.004).면역조직화학시CD40、CD40L화MMP9적양성표체면적A조대우B조(FCD40=39.451,FCD40L=57.606,FMMP9=30.711,P=0.000),B조대우C조(FCD40=95.066,FCD40L=59.081,FMMP9=145.758,P=0.000);CD40、CD40L여MMP9적양성표체면적유정상관관계(rCD40L-CD40L=0.963,rCD40-MMP9=0.959,rCD40L-MMP9=0.929,P=0.000),병차CD40、CD40L화MMP9균재반괴견부표체명현증다.결론 CD40-CD40L재동맥죽양경화적형성화반괴은정성중기중요작용;기가능통과상조MMP9도치반괴불은정;CD40L가능시통과전록후수식영향CD40L적표체,종이산생생물학효응.
Objective To investigate the expression of CD40,CD40L and MMP9 in carotid atherosclerotic plaques and evaluate their roles in carotid atherosclerotic plaque stability.Methods Carotid atherosclerotic plaques were isolated in carotid eversion endarterectomy (GEE) in 37 patients with high-grade stenosis (>70%) including 20 stroke (A group) and 17 non-stroke patients (B group).The control group included samples of normal carotid artery from 11 normal individuals,The RNA expression levels of CD40,CD40 L and MMP9 in all A,B and control groups were quantitatively detected by real-time quantification polymerase chain reaction (PCR) and the protein expression levels were detected by Western blotting analysis.The expression and distribution of CD40,CD40L and MMP9 in carotid atherosclerotic plaques were detected by immunohistochemistry staining.Then correlations between CD40-CD40L and MMP9 were statistically analyzed.Results The relative CD40 mRNA level in high-grade stenosis of A group,B group and normal control were 2.41±0.43,1.03±0.38 and 0.31±0.12,respectively,and MMP9 mRNA 6.88±1.57,1.90±0.44 and 0.39±0.12,respectively.The levels of CD40 and MMP9 mRNA in A group were significantly higher than those in B group (P=0.000),the levels of CD40 and MMP9 in B group were significantly higher than those in controls (P=0.000).There was a linear correlation between CD40 and MMP9 mRNA (r=0.929,P=0.000).However,there were no significantly difference in mRNA levels of CD40L between carotid atherosclerosis and controls.The protein expression levels of CD40,CD40L and MMP9 in A group were significantly higher than those in B group (FCD40=104.100,P=0.000;FCD40L=129.932,P=0.000;FMMP9=13.565,P=0.021) and B group higher than normal controls (FCD40=115.848,P = 0.000;FCD40L= 30.482,P=0.005;FMMP9=35.557,P=0.004).The areas of positive staining of CD40,CD40L and MMP9 in immunochemistry study in A group were significantly higher than those in B group and B group was significantly higher than controls.There were linear correlations between positive staining areas Of CD40 and CD40L,CD40 and MMP9,CD40L and MMP9 (r=0.963,0.959,0.929,P=0.000).Expressions of CD40,CD40L and MMP9 were significantly higher in the shoulder areas of the atherosclerotic plaques than in other areas.Conclusions The CD40-CD40L has an important role in the formation of carotid atherosclerosis and plaque instability,probably by up-regulating MMP9.The expression of CD40L may be regulated by post-transcriptional modification to exert biological effects.