CAJ | 학술논문

目的探讨基质金属蛋白酶9(MMP-9)在慢性酒精中毒性肌损伤(chronic alcoholic muscle injury,CAMI)发病过程中的作用.方法 60只雄性SD大鼠随机分为对照组(A组,15只,生理盐水灌胃)和实验组(B组,45只,灌酒),建立CAMI模型.将A组再随机分为A1～3组(分别于生理盐水灌胃第2周、第6周和第12周末处死,每组5只);B组再随机分为B1～3组(分别于灌酒第2周、第6周和第12周末处死,每组15只).光镜下观察大鼠骨骼肌病理学改变;逆转录-聚合酶链反应检测大鼠骨骼肌MMP-9的mRNA表达的动态改变;Western blot检测大鼠骨骼肌MMP-9蛋白的表达变化;明胶酶谱法检测大鼠骨骼肌MMP-9的酶活性变化.结果组织化学结果显示,B组大鼠跖肌Ⅱ型肌纤维萎缩,间质炎性细胞浸润和纤维增生,其中I型胶原增生明显.A1组与B1组大鼠骨骼肌MMP-9 mRNA的表达甚少,Western blot和酶谱法检测仅显示MMP-9前体型.B2组大鼠跖肌MMP-9 mRNA表达(0.0579±0.0046)较A2组(0.0198±0.0023)增高(t=-9.151,P<0.01),B3组大鼠跖肌和腓肠肌MMP-9 mRNA的表达(0.1417±0.0116、0.0599±0.0057)均较A3组(0.0249±0.0049、0.0237±0.0021)增高(t=-18.095,P<0.01;t=-5.933,P<0.05).同时,Western blot和酶谱法也检测出B3组大鼠腓肠肌和跖肌活性型MMP-9的表达量及酶活性进一步增强,与B2组相比差异有统计学意义.结论 MMP-9的表达和活性增高可能参与了骨骼肌间质纤维增生,促进了CAMI.
목적 탐토기질금속단백매9(MMP-9)재만성주정중독성기손상(chronic alcoholic muscle injury,CAMI)발병과정중적작용.방법 60지웅성SD대서수궤분위대조조(A조,15지,생리염수관위)화실험조(B조,45지,관주),건립CAMI모형.장A조재수궤분위A1～3조(분별우생리염수관위제2주、제6주화제12주말처사,매조5지);B조재수궤분위B1～3조(분별우관주제2주、제6주화제12주말처사,매조15지).광경하관찰대서골격기병이학개변;역전록-취합매련반응검측대서골격기MMP-9적mRNA표체적동태개변;Western blot검측대서골격기MMP-9단백적표체변화;명효매보법검측대서골격기MMP-9적매활성변화.결과 조직화학결과현시,B조대서척기Ⅱ형기섬유위축,간질염성세포침윤화섬유증생,기중I형효원증생명현.A1조여B1조대서골격기MMP-9 mRNA적표체심소,Western blot화매보법검측부현시MMP-9전체형.B2조대서척기MMP-9 mRNA표체(0.0579±0.0046)교A2조(0.0198±0.0023)증고(t=-9.151,P<0.01),B3조대서척기화비장기MMP-9 mRNA적표체(0.1417±0.0116、0.0599±0.0057)균교A3조(0.0249±0.0049、0.0237±0.0021)증고(t=-18.095,P<0.01;t=-5.933,P<0.05).동시,Western blot화매보법야검측출B3조대서비장기화척기활성형MMP-9적표체량급매활성진일보증강,여B2조상비차이유통계학의의.결론 MMP-9적표체화활성증고가능삼여료골격기간질섬유증생,촉진료CAMI.
Objective To investigate the effects of matrix metslloproteinase 9(MMP-9)on chronic alcohol muscle injury(CAMI).Methods Sixty male Sprague-Dawley(SD)rats were randomly divided into control group(A group,15 rats)and experimental group(B group,45 rats).All rats were administered intragagtfically by gavage.Group A was treated with physiological saline.Group B was treated with alcohoL Initial dose was 4 g·kg~(-1)·d~(-1).Then dose wag increased by 2 g·kg~(-1)·d~(-1)weekly until 10 g·kg~(-1)·d~(-1).Group A were randomly divided into 3 subset groups(A1-3),which would be sacrificed at 2,6 and 12 weeks after feeding physiological saline.Each group had 5 rats.Group B were randomly divided into 3 subset groups(B1-3),which would be sacrificed at 2,6 and 12 weeks after feeding alcohol.Each group had 15 rats.Then the histopathological changes in rat skeletal muscles were observed by HE and NADH-TR staining.The expression of MMP-9 mRNA was detected by RT-PCR MMP-9 protein was detected by Western blot The enzymatic activity of MMP-9 weag detected by gelatin zymography.Results The observation of histechemistry showed that there were obviously atrophic type-Ⅱmuscle fibers of plantaris in the B group rats,infiltration of inflammatory cells and accrementition of connective tissue between muscles.in which type-I collagen was obviously proliferative.The expression of MMP-9 mRNA in group A1 and group B1 was much lower.The precursor of MMP-9 in group AI and group B1 wag detected by Western blot and gelatin zymography.The mRNA expression of MMP-9 of plantaris in the group B2(0.0579±0.0046)was significantly elevated than that in the group A2(0.0198±0.0023,t=-9.151,P<0.01).The mRNA expression of MMP-9 of plantaris and gastrocnemius in the group B3(0.1417±0.0116,0.0599±0.0057)was significantly higher than that in the group A3(0.0249±0.0049,t=-18.095,P<0.01;0.0237±0.0021,t=-5.933,P<0.05).Meanwhile,the expression and enzymatic activity of MMP-9 of nJantaris and gastroenemius in the group B2 Were detected by Western blot and gelatin zymography.The expression and enzymatic activity of M MP-9 of plantaris and gastrocnemius in the group B3 were significantly inereased compared with the group B2.Conclusions The inereased expression and the increased enzymatic activity of MMP-9 may be involved in interstitial fibrosis of CAMI and aggravate the development of CAMI.