摘要
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.
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.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第8期564-567,共4页
National Medical Journal of China
基金
基金项目：国家自然科学基金（81070660）
广东省自然科学重点资助项目（10251008901000010）
