中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2012年
10期
1106-1110
,共5页
薛声能%雷娟%杨川%林刁珠%严励
薛聲能%雷娟%楊川%林刁珠%嚴勵
설성능%뢰연%양천%림조주%엄려
基质金属蛋白酶9%成纤维细胞%生物学行为%增殖%细胞活力%分泌%迁移%糖尿病足
基質金屬蛋白酶9%成纖維細胞%生物學行為%增殖%細胞活力%分泌%遷移%糖尿病足
기질금속단백매9%성섬유세포%생물학행위%증식%세포활력%분비%천이%당뇨병족
Matrix metalloproteinase 9%Fibroblast%Biological behaviors%Proliferation%Cell viability%Secretion%Migration%Diabetic foot
目的 观察基质金属蛋白酶9(matrixmetalloproteinase 9,MMP 9)高表达对大鼠皮肤成纤维细胞生物学行为的影响,以阐述其在糖尿病足伤口愈合中的可能机制.方法 本研究在中山大学孙逸仙纪念医院内分泌专科实验室完成.大鼠皮肤成纤维细胞与高糖(22 mmol/L)+高同型半胱氨酸(100 μmol/L)联合孵育建立体外MMP 9高表达细胞模型,另设正糖(5.5 mmol/L)孵育对照组.Realtime PCR、ELISA和明胶酶谱法检测MMP 9 mRNA、蛋白表达量及活性,流式细胞仪检测细胞增殖功能,CCK-8检测细胞活力,ELISA法检测细胞胶原(羟脯氨酸)分泌能力、划痕实验评价细胞横向迁移能力、Transwell法评价细胞纵向迁移能力.计量数据以均数±标准差(x-±s)表示,两组间均数的比较采用成组t检验,以P ＜0.05为差异具有统计学意义.结果 与对照组比较,MMP 9高表达组大鼠皮肤成纤维细胞的MMP 9 mRNA、蛋白表达量及活性,分别升高6.05倍、4.12倍和1.58倍(P＜0.01)；与此同时MMP 9高表达组大鼠皮肤成纤维细胞S期的比例、增殖指数、细胞活力、胶原(羟脯氨酸)分泌量、6h横向迁移率和纵向迁移细胞数,分别下降29.8％、18.1％、23.3％、68.7％、45.0％和21.4％ (P ＜0.01).结论 MMP 9高表达的皮肤成纤维细胞增殖减慢、活力下降、迁移和胶原分泌能力降低,提示MMP 9可能抑制成纤维细胞的生物学行为.
目的 觀察基質金屬蛋白酶9(matrixmetalloproteinase 9,MMP 9)高錶達對大鼠皮膚成纖維細胞生物學行為的影響,以闡述其在糖尿病足傷口愈閤中的可能機製.方法 本研究在中山大學孫逸仙紀唸醫院內分泌專科實驗室完成.大鼠皮膚成纖維細胞與高糖(22 mmol/L)+高同型半胱氨痠(100 μmol/L)聯閤孵育建立體外MMP 9高錶達細胞模型,另設正糖(5.5 mmol/L)孵育對照組.Realtime PCR、ELISA和明膠酶譜法檢測MMP 9 mRNA、蛋白錶達量及活性,流式細胞儀檢測細胞增殖功能,CCK-8檢測細胞活力,ELISA法檢測細胞膠原(羥脯氨痠)分泌能力、劃痕實驗評價細胞橫嚮遷移能力、Transwell法評價細胞縱嚮遷移能力.計量數據以均數±標準差(x-±s)錶示,兩組間均數的比較採用成組t檢驗,以P ＜0.05為差異具有統計學意義.結果 與對照組比較,MMP 9高錶達組大鼠皮膚成纖維細胞的MMP 9 mRNA、蛋白錶達量及活性,分彆升高6.05倍、4.12倍和1.58倍(P＜0.01)；與此同時MMP 9高錶達組大鼠皮膚成纖維細胞S期的比例、增殖指數、細胞活力、膠原(羥脯氨痠)分泌量、6h橫嚮遷移率和縱嚮遷移細胞數,分彆下降29.8％、18.1％、23.3％、68.7％、45.0％和21.4％ (P ＜0.01).結論 MMP 9高錶達的皮膚成纖維細胞增殖減慢、活力下降、遷移和膠原分泌能力降低,提示MMP 9可能抑製成纖維細胞的生物學行為.
목적 관찰기질금속단백매9(matrixmetalloproteinase 9,MMP 9)고표체대대서피부성섬유세포생물학행위적영향,이천술기재당뇨병족상구유합중적가능궤제.방법 본연구재중산대학손일선기념의원내분비전과실험실완성.대서피부성섬유세포여고당(22 mmol/L)+고동형반광안산(100 μmol/L)연합부육건입체외MMP 9고표체세포모형,령설정당(5.5 mmol/L)부육대조조.Realtime PCR、ELISA화명효매보법검측MMP 9 mRNA、단백표체량급활성,류식세포의검측세포증식공능,CCK-8검측세포활력,ELISA법검측세포효원(간포안산)분비능력、화흔실험평개세포횡향천이능력、Transwell법평개세포종향천이능력.계량수거이균수±표준차(x-±s)표시,량조간균수적비교채용성조t검험,이P ＜0.05위차이구유통계학의의.결과 여대조조비교,MMP 9고표체조대서피부성섬유세포적MMP 9 mRNA、단백표체량급활성,분별승고6.05배、4.12배화1.58배(P＜0.01)；여차동시MMP 9고표체조대서피부성섬유세포S기적비례、증식지수、세포활력、효원(간포안산)분비량、6h횡향천이솔화종향천이세포수,분별하강29.8％、18.1％、23.3％、68.7％、45.0％화21.4％ (P ＜0.01).결론 MMP 9고표체적피부성섬유세포증식감만、활력하강、천이화효원분비능력강저,제시MMP 9가능억제성섬유세포적생물학행위.
Objective To study the effects of high matrix metalloproteinase 9 (MMP 9 ) on the biological behaviors of fibroblasts,in order to clarify the possible mechanisms in the wound healing of diabetic foot.Methods Establish the cell model of skin fibroblast with high expression of MMP 9 by high glucose (22.0 mmol/L) and high homocysteine (100 μmol/L) co-culture.Control group was incubated with normal glucose (5.5 mmol/L).Realtime PCR,ELISA and gelatin zymography were used to detect the MMP 9 mRNA,protein expression and activity of MMP 9.Flow cytometry,CCK-8,ELISA assay,scratch test and transwell were used to detect cell proliferation,viability,collagen (hydroxyproline) secretion,horizontal migration and vertical migration of cells.Results Data were expressed as (x- ± s).Differences were considered significant if their P value was ＜ 0.05.Results The expression of MMP 9 mRNA,protein levels and the activity of MMP 9 in high MMP 9 group were 6.05 folds,4.12 folds and 1.58 folds higher than those in control group (P ＜ 0.01,respectively).The proportion of S phase cells,proliferation index,cell viability,collagen (hydroxyproline) secretion,6 h horizontal migration rate and the number of vertical migration cells in high MMP9 group decreased by 29.8 ％,18.1％,23.3 ％,68.7 ％,45.0 ％ and 21.4 ％ than those in control group (P ＜ 0.01,respectively ). Conclusions Fibroblast with high expression of MMP 9 have decreased proliferation,activity,collagen secretion and reduced migration,which suggests that MMP 9 may inhibit the biological behaviors of fibroblasts.
