【摘要】：背景:真皮成纖維細(xì)胞是最主要的皮膚創(chuàng)傷修復(fù)細(xì)胞,能產(chǎn)生膠原及金屬蛋白酶等。明膠酶(包括MMP-2和MMP-9)能降解IV型膠原及細(xì)胞外基質(zhì),在皮膚傷口愈合過程中發(fā)揮重要作用。我們之前的研究已經(jīng)證實(shí)了凝血酶可激活真皮成纖維細(xì)胞釋放MMP-9。而真皮成纖維細(xì)胞在凝血酶與炎癥因子IL-1β、TNF-α單獨(dú)或聯(lián)合激發(fā)下釋放MMP-2和MMP-9的情況,目前仍知之甚少。 目的:研究凝血酶和炎癥因子IL-1β、TNF-α激發(fā)人真皮成纖維細(xì)胞釋放MMP-2和MMP-9的情況。 方法:體外培養(yǎng)成人原代真皮成纖維細(xì)胞,單獨(dú)或聯(lián)合加入凝血酶5u/ml,IL-1β5ng/ml、10ng/ml和TNF-α5ng/ml、10ng/ml,經(jīng)過2h、6h、24h激發(fā)后用明膠酶譜法檢測真皮成纖維細(xì)胞釋放MMP-2和MMP-9的活性。 結(jié)果:(1)在不同的時(shí)間點(diǎn)(2h,6h,24h),凝血酶5u/ml均可增強(qiáng)真皮成纖維細(xì)胞釋放MMP-9的活性,對MMP-2活性影響不明顯。IL-1β5ng/ml、10ng/ml和TNF-α5ng/ml、10ng/ml在6h和24h的時(shí)間位點(diǎn)可增強(qiáng)真皮成纖維細(xì)胞釋放MMP-9的活性,對MMP-2活性影響不明顯。隨著刺激時(shí)間的延長,凝血酶、IL-1β、TNF-α激發(fā)成纖維細(xì)胞釋放MMP-9的活性水平逐漸升高,呈時(shí)間依賴性。(2)在不同的時(shí)間點(diǎn)(2h,6h,24h),凝血酶5u/ml與IL-1β10ng/ml聯(lián)合作用,激發(fā)成纖維細(xì)胞釋放MMP-9的活性水平高于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05);在2h的時(shí)間點(diǎn),凝血酶5u/ml與TNF-α10ng/ml聯(lián)合作用,成纖維細(xì)胞釋放MMP-9的活性水平低于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05);在6h和24h的時(shí)間點(diǎn),凝血酶5u/ml與TNF-α10ng/ml聯(lián)合作用,成纖維細(xì)胞釋放MMP-9的活性水平高于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。(3)在不同的時(shí)間點(diǎn)(2h,6h,24h),凝血酶5ng/ml與IL-1β10ng/ml、TNF-α10ng/ml三者聯(lián)合作用,成纖維細(xì)胞釋放MMP-9活性水平低于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),MMP-2活性水平無明顯變化。IL-1β10ng/ml與TNF-α10ng/ml聯(lián)合作用,成纖維細(xì)胞釋放MMP-9水平無明顯變化,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論:(1)凝血酶和炎癥因子(IL-1β、TNF-α)能激發(fā)成人真皮成纖維細(xì)胞釋放MMP-9增強(qiáng),對MMP-2作用不明顯;隨著刺激時(shí)間的延長,凝血酶、IL-1β、TNF-α激活成纖維細(xì)胞釋放MMP-9的水平逐漸升高,呈時(shí)間依賴性。(2)凝血酶單獨(dú)與IL-1β或TNF-α作用,能激發(fā)成纖維細(xì)胞釋放MMP-9增強(qiáng),但凝血酶與IL-1β、TNF-α三者聯(lián)合作用使成纖維細(xì)胞釋放MMP-9降低。(3)IL-1β與TNF-α聯(lián)合作用,對成纖維細(xì)胞釋放MMP-9無變化。
[Abstract]:Background: dermal fibroblasts are the most important wound repair cells, producing collagen and metalloproteinases. Gelatinases (including MMP-2 and MMP-9) can degrade collagen type IV and extracellular matrix and play an important role in skin wound healing. Our previous studies have shown that thrombin activates dermal fibroblast release of MMP-9. However, little is known about the release of MMP-2 and MMP-9 by dermal fibroblasts stimulated by thrombin alone or in combination with inflammatory cytokines IL-1 尾 and TNF- 偽. Aim: to study the effects of thrombin and inflammatory factor IL-1 尾, TNF- 偽 on the release of MMP-2 and MMP-9 from human dermal fibroblasts. Methods: adult primary dermal fibroblasts were cultured alone or in combination with thrombin 5u / ml IL-1 尾 5ng / ml and TNF- 偽 5ng / ml 10ng / ml. The activity of MMP-2 and MMP-9 in dermal fibroblasts was detected by gelatinase assay after 24 h stimulation. Results: (1) at different time points (2 h, 6 h and 24 h), thrombin 5u/ml could enhance the activity of MMP-9 released by dermal fibroblasts, but had no significant effect on MMP-2 activity. At the time point of 6 h and 24 h, 10ng/ml could enhance the activity of MMP-9 release from dermal fibroblasts, but had no obvious effect on MMP-2 activity. With the prolongation of stimulation time, the activity of thrombin, IL-1 尾 and TNF- 偽 stimulated by fibroblasts to release MMP-9 increased in a time dependent manner. (2) at different time points (2 h, 6 h, 24 h), the activity of thrombin, IL-1 尾 and TNF- 偽 increased in a time-dependent manner. Combined effect of thrombin 5u/ml and IL-1 尾 10ng/ml stimulated the activity of MMP-9 release from fibroblasts, which was significantly higher than that in the control group (P0.05). At the time point of 2h, the activity of MMP-9 released by fibroblasts was lower than that of the control group (P0.05) when thrombin 5u/ml combined with TNF- 偽 10ng/ml. At the time points of 6 h and 24 h, the activity of MMP-9 released by fibroblasts in combination of thrombin 5u/ml and TNF- 偽 10ng/ml was significantly higher than that in the control group (P0.05). (3) at different time points (2 h ~ 6 h). 24 h), combined with thrombin 5ng/ml and IL-1 尾 10 ng / ml TNF- 偽 10ng/ml, the activity of MMP-9 released by fibroblasts was significantly lower than that in control group (P0.05). There was no significant change in the activity of MMP-2. The level of MMP-9 released by fibroblasts did not change in combination of IL-1 尾 10ng/ml and TNF- 偽 10ng/ml (P0.05). Conclusion: (1) Thrombin and IL-1 尾 (TNF- 偽) can stimulate the increase of MMP-9 release from adult dermal fibroblasts, but the effect on MMP-2 is not obvious. With the prolongation of stimulation time, the level of MMP-9 released by fibroblasts activated by thrombin, IL-1 尾 and TNF- 偽 gradually increased in a time dependent manner. (2) thrombin acted alone with IL-1 尾 or TNF- 偽. The fibroblast release of MMP-9 was enhanced, but the combination of thrombin and IL-1 尾, TNF- 偽 decreased the release of MMP-9. (3) the combination of IL-1 尾 and TNF- 偽. There was no change in the release of MMP-9 from fibroblasts.
【學(xué)位授予單位】：汕頭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R751

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