CAJ | 학술논문

目的探讨基质金属蛋白酶9( MMP-9)对皮肤成纤维细胞增殖、凋亡及合成Ⅰ、Ⅲ型前胶原的影响.方法通过0.1、1.0μg/n1脂多糖(LPS)刺激大鼠皮肤成纤维细胞18、48 h建立MMP-9高表达模型；通过RNA干扰(RNAi)技术抑制模型MMP-9的表达.分别观察MMP-9高表达模型和RNAi技术抑制MMP-9表达后成纤维细胞增殖、凋亡和合成Ⅰ、Ⅲ型前胶原的改变.活细胞计数试剂盒(CCK)8检测细胞增殖；流式细胞技术检测细胞凋亡；实时定量逆转录PCR(qRT-PCR)检测Ⅰ、Ⅲ型前胶原基因表达.结果 1.0 μg/ml LPS干预成纤维细胞48 h,MMP-9表达明显高于对照组,mRNA表达量为对照组的14.25倍,蛋白表达量为对照组的2.31倍(均P＜0.05).MMP-9高表达模型在抑制MMP-9高表达后MMP-9 mRNA表达量为模型对照组的1/8,蛋白表达量为模型对照组的1/3(均P＜0.05).MMP-9表达增高伴有皮肤成纤维细胞增殖减慢[(1.08±0.08)比(1.18±0.09),P＜0.05],抑制MMP-9高表达后细胞增殖明显改善[(1.78±0.17)比 (1.53±0.15),P＜0.01].MMP-9高表达后成纤维细胞凋亡比例与对照组比较差异无统计学意义(P＞0.05),抑制MMP-9高表达后48 h细胞凋亡比例(3.53％±0.22％)低于模型对照组(4.47％±0.46％,P=0.033).MMP-9高表达及抑制高表达后对成纤维细胞Ⅰ型胶原合成均无明显影响(均P＞0.05)；MMP-9表达增高伴有Ⅲ型前胶原合成减少(P＜0.01),但抑制MMP-9高表达后未见Ⅲ型前胶原mRNA表达增多(P＞0.05).结论 MMP-9可影响皮肤成纤维细胞增殖等生物学行为.
목적 탐토기질금속단백매9( MMP-9)대피부성섬유세포증식、조망급합성Ⅰ、Ⅲ형전효원적영향.방법통과0.1、1.0μg/n1지다당(LPS)자격대서피부성섬유세포18、48 h건립MMP-9고표체모형；통과RNA간우(RNAi)기술억제모형MMP-9적표체.분별관찰MMP-9고표체모형화RNAi기술억제MMP-9표체후성섬유세포증식、조망화합성Ⅰ、Ⅲ형전효원적개변.활세포계수시제합(CCK)8검측세포증식；류식세포기술검측세포조망；실시정량역전록PCR(qRT-PCR)검측Ⅰ、Ⅲ형전효원기인표체.결과 1.0 μg/ml LPS간예성섬유세포48 h,MMP-9표체명현고우대조조,mRNA표체량위대조조적14.25배,단백표체량위대조조적2.31배(균P＜0.05).MMP-9고표체모형재억제MMP-9고표체후MMP-9 mRNA표체량위모형대조조적1/8,단백표체량위모형대조조적1/3(균P＜0.05).MMP-9표체증고반유피부성섬유세포증식감만[(1.08±0.08)비(1.18±0.09),P＜0.05],억제MMP-9고표체후세포증식명현개선[(1.78±0.17)비 (1.53±0.15),P＜0.01].MMP-9고표체후성섬유세포조망비례여대조조비교차이무통계학의의(P＞0.05),억제MMP-9고표체후48 h세포조망비례(3.53％±0.22％)저우모형대조조(4.47％±0.46％,P=0.033).MMP-9고표체급억제고표체후대성섬유세포Ⅰ형효원합성균무명현영향(균P＞0.05)；MMP-9표체증고반유Ⅲ형전효원합성감소(P＜0.01),단억제MMP-9고표체후미견Ⅲ형전효원mRNA표체증다(P＞0.05).결론 MMP-9가영향피부성섬유세포증식등생물학행위.
Objective To explore the effects of matrix metallopeptidase 9 (MMP-9) on the proliferation, apoptosis, type Ⅰ and Ⅲ procollagen synthesis of rat dermal fibroblasts. Methods Lipopolysaccharide (LPS) at 0.1 and 1.0,μg/ml was used to stimulate fibroblasts to up-regulate the expression of MMP-9 for 18 and 48 h. And then RNA interference was employed to inhibit the high expression of MMP-9.Cell proliferation was tested by CCK-8,cell apoptosis by flow cytometry and type Ⅰ and Ⅲ procollagen expressions by quantitative reverse transcriptase PCR (qRT-PCR). Results The MMP-9 expression of fibroblasts increased after the stimulation of LPS.And the 1.0 μg/ml LPS stimulation of 48 hours was 14.25 times of mRNA expression and 2.31 times of protein expression versus that of the normal group (both P ＜ 0.05 ).The RNA interference obviously inhibited the high expression of MMP-9.The mRNA expression was 1/8 and protein expression 1/3 ( both P ＜ 0.05) as compared with the control group.Cell proliferation decreased with the rising expression of MMP-9 to some extent[( 1.08 ±0.08) vs ( 1.18 ±0.09),P ＜0.05]and improved after the inhibition of high expression of MMP-9[(1.78 ±0.17) vs (1.53 ±0.15),P ＜0.01].There was no change of apoptosis accompanied with high expression of MMP-9 (P ＞ 0.05 ).Apoptosis decreased after the inhibition of high expression of MMP-9 for 48 hours ( 3.53％ ± 0.22％ vs 4.47％ ±0.46％,P ＜0.05).The synthesis of type Ⅰ proeollagen was the same no matter up-regulation or down-regulation of MMP-9 expression (P ＞ 0.05).As the expression of MMP-9 increased,the synthesis of type Ⅲ procollagen decreased (P ＜0.01 ),but not increased by 1.02 folds after the inhibited expression of MMP-9 (P ＞ 0.05).Conclusion MMP-9 can affect the biological behaviors of rat dermal fibroblasts.