CAJ | 학술논문

目的研究肝细胞生长因子(HGF)对血管紧张素Ⅱ(AngⅡ)诱导下新生大鼠心肌成纤维细胞胶原合成降解平衡的影响.方法差速贴壁法分离新生SD大鼠心肌成纤维细胞(CF),并用细胞免疫组织化学进行鉴定.实验分为正常对照组(C组)、Ang II 10-6mol/L组(A组)、HGF 10μg,L+AngⅡ10-6 mol/L组(H1组)、HGF 100μg/L+AngⅡ 10-6 mol/L组(H2组).作用48 h后采用羟脯氨酸法测胶原蛋白含量,RT-PCR法测Ⅰ型胶原(Col Ⅰ)、Ⅲ型胶原(Col Ⅲ)、基质金属蛋白酶-1(MMP-1)、基质金属蛋ca酶抑制因子-1(TIMP-1)mRNA的表达;Westernblotting 4col Ⅰ蛋白水平的表达.用Ⅰ型胶原酶活性检测试剂盒测MMP-1的活性.结果作用48 h后,A组、H1组胶原蛋白含量较C组增加[(39.08±2.71)mg/L比(37.45±4.22)mg/L比(23.73±1.62)ms/L,均P＜0.05],H2组胶原蛋白含量(26.03±3.04)mg/L则低于A组、Hl组(P＜0.05),与C组差异无统计学意义.在mRNA水平上ColⅠ、col Ⅲ、TIMP-1的表达A组＞H1组＞H2组＞C组(均P＜0.05),而MMP-1 mRNA表达A组＜H1组＜H2组＜C组(均P＜0.05).Col Ⅰ蛋白水平A组＞H1组＞H2组＞C组(89.90±14.29比68.21±11.43比36.08±8.8比30.14±7.36,均P＜0.05).A组、H1组MMP-1 mRNA表达量及活性水平较C组明显降低(均P＜0.05),H2组则基本恢复到C组水平.结论 HGF主要通过激活胶原降解途径,重调MMP-1.TIMP-1平衡,逆转心肌纤维化.
목적 연구간세포생장인자(HGF)대혈관긴장소Ⅱ(AngⅡ)유도하신생대서심기성섬유세포효원합성강해평형적영향.방법 차속첩벽법분리신생SD대서심기성섬유세포(CF),병용세포면역조직화학진행감정.실험분위정상대조조(C조)、Ang II 10-6mol/L조(A조)、HGF 10μg,L+AngⅡ10-6 mol/L조(H1조)、HGF 100μg/L+AngⅡ 10-6 mol/L조(H2조).작용48 h후채용간포안산법측효원단백함량,RT-PCR법측Ⅰ형효원(Col Ⅰ)、Ⅲ형효원(Col Ⅲ)、기질금속단백매-1(MMP-1)、기질금속단ca매억제인자-1(TIMP-1)mRNA적표체;Westernblotting 4col Ⅰ단백수평적표체.용Ⅰ형효원매활성검측시제합측MMP-1적활성.결과 작용48 h후,A조、H1조효원단백함량교C조증가[(39.08±2.71)mg/L비(37.45±4.22)mg/L비(23.73±1.62)ms/L,균P＜0.05],H2조효원단백함량(26.03±3.04)mg/L칙저우A조、Hl조(P＜0.05),여C조차이무통계학의의.재mRNA수평상ColⅠ、col Ⅲ、TIMP-1적표체A조＞H1조＞H2조＞C조(균P＜0.05),이MMP-1 mRNA표체A조＜H1조＜H2조＜C조(균P＜0.05).Col Ⅰ단백수평A조＞H1조＞H2조＞C조(89.90±14.29비68.21±11.43비36.08±8.8비30.14±7.36,균P＜0.05).A조、H1조MMP-1 mRNA표체량급활성수평교C조명현강저(균P＜0.05),H2조칙기본회복도C조수평.결론 HGF주요통과격활효원강해도경,중조MMP-1.TIMP-1평형,역전심기섬유화.
Objective To study the effects of hepatocyte growth factor (HGF) on angiotensin Ⅱ(Ang Ⅱ )-induced synthesis and degradation of cardiac fibroblast collagen in neonatal mice.Methods Cardiac fibroblasts (CFs) of neonatal Sprague-Dawley (SD) rats were isolated by differential adhesion, and were identified by immunocytochemistry.The cells were assigned to normal control group (group C) , 10-6 mol/L Ang Ⅱ group (group A), 10μg/L HGF + 10-6 mol/L Ang Ⅱ group (group H1) and 100μg/L HGF +10-6 mol/L Ang Ⅱ group (group H2).All the groups were treated for 48 h.Hydroxyproline assay was used to determine the collagen protein level.RT-PCR was used to detect the expression of ColⅠ, ColⅢ, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-l) mRNA, and Western blotting was employed to measure Col I protein.In addition, MMP-1 activity was evaluated by collagenase Ⅰ activity assay kit.Results After intervention for 48 h, the collagen protein levels in groups A and H1 were significantly higher compared with group C [ (39.08±2.71) mg/L vs (37.45±4.22) mg/L vs (23.73 ±±1.62) mg/L, all P＜0.05] and the collagen protein level in group H2 [(26.03±3.04) mg/L] was lower than those of groups A and Hl(P＜0.05), but was not statistically different from group C.The expression levels of Col Ⅰ ,Col M and TIMP-1 showed an order of group A＞group HI＞group H2> group C (all P＜0.05), and for MMP-1 mRNA, group A ＜ group H1 ＜ group H2 ＜ group C (all P＜0.05).The levels of Col I protein expression showed an order of group A＞group HI＞group H2＞group C (89.90±4.29 vs 68.21?1.43 vs 36.08?.8 vs 30.14?.36, all P＜0.05).The expression and activity of MMP-1 mRNA in group A and H1 were obviously lower than those in group C (all P＜0.05), while group H2 were comparable to group C in these data.Conclusion HGF re-regulates MMP-1-TIMP-1 balance and ameliorates myocardiac fibrosis mainly by activating collagen degradation.