糖尿病足部溃疡患者白介素-1β对成纤维细胞增殖及基质金属蛋白酶-2和基质金属蛋白酶-9的影响

目的检测糖尿病足部溃疡创面白介素-1β（IL-1β）的含量，探讨IL-1β对溃疡局部成纤维细胞（Fb）增殖功能及其基质金属蛋白酶-2（MMP-2），基质金属蛋白酶-9（MMP-9）活性和其蛋白表达的影响。方法取糖尿病足溃疡边缘成纤维细胞培养、传代至4-6代，在24、48、72h时间段分别加培养液及不同浓度IL—1β，观察成纤维细胞生长。并用酶谱法检测IL-1β对真皮成纤维细胞MMP-2、MMP-9活性的影响，用蛋白印迹（Westernblott）检测其MMP-2、MMP-9蛋白表达量。结果糖尿病急性创面组IL-1β含量[（8．84±7．06）ng／m1]低于糖尿病足部难治溃疡组[（89．16±38．14）ng／m1]，差别有统计学意义（P〈0．001）；IL-1β0．5～5ng／ml对Fb增殖有明显促进作用，随着IL-1p浓度增加，对F1）的促增殖作用减弱，IL-1β50ng／ml对Fb增殖无影响；IL-1β500ng／ml对Fb增殖有明显抑制效应（P〈0．01）；IL-1β50ng／ml不仅能显著增加基质金属蛋白酶（MMP-2，MMP-9）的活性，而且也使MMP-2和MMP-9蛋白的表达显著增加。结论糖尿病足难治溃疡组IL-1β显著增高，进而抑制了成纤维细胞的增殖，同时通过增加MMP-2、MMP-9活性及其蛋白表达，使细胞外基质等分解增加。

The Interaction between lnterin-1β and Fibroblast Proliferation, MMP-2, MMP-9 in Patients with Diabetic Foot Ulcers

Objective Abstract： Objective To inquire into the reason that chronic diabetic foot ulcer （CDF） was difficult to be cured by clarifying their interaction between interin-1β （IL-1β） and fibroblast proliferation, the activity and protein expression of matrix metalloproteinase-2 （MMP-2）, matrix metalloproteinase-9 （MMP-9）in fibroblasts from CDF. Method Fibroblast collected from the edge area of the diabetic foot ulcer was cultured for 4-6 generations, and added in with different concentration of IL-1β at the 24 hr, 48 hr and 72 hr, then the effect of IL-1β on the activity and expression of MMP-2 and MMP-9 in fibroblasts were observed by methods of zymography and Western blott. Results There was a significant difference of IL-1β between CDF and acute diabetic lesions （ADL）; IL-1β in 0.5 ng/ml-5 ng/ml could promote fibroblast proliferation obviously, but weakened gradually with increasing of the IL-1β, but no affection on fibroblast proliferation when IL-1β was in 50 ng/ml, and inhabited fibroblast proliferation over 50 ng/ml of IL-1β; IL-1β in 50 ng/ml increased the activity and protein expression of MMP-2 and MMP-9 in fibroblasts from CDF. Conclusion The reason that CDF was difficult to be cured were related to significant increase of IL-1β. The increase of IL-1β not only inhibits the fibroblast proliferation, but also promotes the activity and protein expression of MMP-2 ＆ MMP-9, and it results in the increase of extraceUular matrix degradation.