生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2014年
1期
102-104
,共3页
章淑平%骆建民%王俐
章淑平%駱建民%王俐
장숙평%락건민%왕리
明胶酶谱%基质金属蛋白酶%聚丙烯酰胺凝胶
明膠酶譜%基質金屬蛋白酶%聚丙烯酰胺凝膠
명효매보%기질금속단백매%취병희선알응효
gelatin zymography%matrix metalloproteinase%polyacrylamide gel
目的：优化明胶酶谱实验方法，并检测不同浓度凝血酶刺激人皮肤成纤维细胞分泌的明胶酶B（MMP-9）活性的变化。方法：用含明胶的聚丙烯酰胺凝胶电泳分开上清中各蛋白条带，经2.5% Triton X-100洗脱、明胶酶缓冲液孵育、考马斯亮蓝染色液染色及脱色液脱色后，经光密度扫描记录，应用ImageJ软件进行密度计量分析。结果：凝血酶刺激人皮肤成纤维细胞分泌MMP-9具有浓度依赖性。结论：利用优化的明胶酶谱实验方法可做出高质量的明胶酶谱图片，对检测明胶酶活性有重要意义。
目的：優化明膠酶譜實驗方法，併檢測不同濃度凝血酶刺激人皮膚成纖維細胞分泌的明膠酶B（MMP-9）活性的變化。方法：用含明膠的聚丙烯酰胺凝膠電泳分開上清中各蛋白條帶，經2.5% Triton X-100洗脫、明膠酶緩遲液孵育、攷馬斯亮藍染色液染色及脫色液脫色後，經光密度掃描記錄，應用ImageJ軟件進行密度計量分析。結果：凝血酶刺激人皮膚成纖維細胞分泌MMP-9具有濃度依賴性。結論：利用優化的明膠酶譜實驗方法可做齣高質量的明膠酶譜圖片，對檢測明膠酶活性有重要意義。
목적：우화명효매보실험방법，병검측불동농도응혈매자격인피부성섬유세포분비적명효매B（MMP-9）활성적변화。방법：용함명효적취병희선알응효전영분개상청중각단백조대，경2.5% Triton X-100세탈、명효매완충액부육、고마사량람염색액염색급탈색액탈색후，경광밀도소묘기록，응용ImageJ연건진행밀도계량분석。결과：응혈매자격인피부성섬유세포분비MMP-9구유농도의뢰성。결론：이용우화적명효매보실험방법가주출고질량적명효매보도편，대검측명효매활성유중요의의。
Objective: To determine the MMP-9 activity in human primary dermal fibroblast(HDF) stimulated by thrombin with the modified gelatin zymography. Methods: The culture medium of HDF was collected stimulat-ed by different dosage of thrombin. The medium was separated by native polyacrylamide gel containing gelatin and incubated with substrate buffer. The MMP-9 digests its substrate of gelatin and shows clear in gel by staining with Coomassie blue. The clear bands were measured and quatified with ScionImage program software. Results:Thrombin provoking the expression of MMP-9 from primary cultured HDF is in a concentration-dependent man-ner. Conclusion: The results indicated that the improved method of gelatin zymography can acquire the high quali-ty images, which is effective to detect metal protease activity.
