凝血酶和炎症因子激发人真皮成纤维细胞释放MMP-2和MMP-9的研究
发布时间：2018-10-31 11:52
【摘要】：背景:真皮成纤维细胞是最主要的皮肤创伤修复细胞,能产生胶原及金属蛋白酶等。明胶酶(包括MMP-2和MMP-9)能降解IV型胶原及细胞外基质,在皮肤伤口愈合过程中发挥重要作用。我们之前的研究已经证实了凝血酶可激活真皮成纤维细胞释放MMP-9。而真皮成纤维细胞在凝血酶与炎症因子IL-1β、TNF-α单独或联合激发下释放MMP-2和MMP-9的情况,目前仍知之甚少。 目的:研究凝血酶和炎症因子IL-1β、TNF-α激发人真皮成纤维细胞释放MMP-2和MMP-9的情况。 方法:体外培养成人原代真皮成纤维细胞,单独或联合加入凝血酶5u/ml,IL-1β5ng/ml、10ng/ml和TNF-α5ng/ml、10ng/ml,经过2h、6h、24h激发后用明胶酶谱法检测真皮成纤维细胞释放MMP-2和MMP-9的活性。 结果:(1)在不同的时间点(2h,6h,24h),凝血酶5u/ml均可增强真皮成纤维细胞释放MMP-9的活性,对MMP-2活性影响不明显。IL-1β5ng/ml、10ng/ml和TNF-α5ng/ml、10ng/ml在6h和24h的时间位点可增强真皮成纤维细胞释放MMP-9的活性,对MMP-2活性影响不明显。随着刺激时间的延长,凝血酶、IL-1β、TNF-α激发成纤维细胞释放MMP-9的活性水平逐渐升高,呈时间依赖性。(2)在不同的时间点(2h,6h,24h),凝血酶5u/ml与IL-1β10ng/ml联合作用,激发成纤维细胞释放MMP-9的活性水平高于对照组,差异有统计学意义(P0.05);在2h的时间点,凝血酶5u/ml与TNF-α10ng/ml联合作用,成纤维细胞释放MMP-9的活性水平低于对照组,差异有统计学意义(P0.05);在6h和24h的时间点,凝血酶5u/ml与TNF-α10ng/ml联合作用,成纤维细胞释放MMP-9的活性水平高于对照组,差异有统计学意义(P0.05)。(3)在不同的时间点(2h,6h,24h),凝血酶5ng/ml与IL-1β10ng/ml、TNF-α10ng/ml三者联合作用,成纤维细胞释放MMP-9活性水平低于对照组,差异有统计学意义(P0.05),MMP-2活性水平无明显变化。IL-1β10ng/ml与TNF-α10ng/ml联合作用,成纤维细胞释放MMP-9水平无明显变化,差异无统计学意义(P0.05)。 结论:(1)凝血酶和炎症因子(IL-1β、TNF-α)能激发成人真皮成纤维细胞释放MMP-9增强,对MMP-2作用不明显;随着刺激时间的延长,凝血酶、IL-1β、TNF-α激活成纤维细胞释放MMP-9的水平逐渐升高,呈时间依赖性。(2)凝血酶单独与IL-1β或TNF-α作用,能激发成纤维细胞释放MMP-9增强,但凝血酶与IL-1β、TNF-α三者联合作用使成纤维细胞释放MMP-9降低。(3)IL-1β与TNF-α联合作用,对成纤维细胞释放MMP-9无变化。
[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- 伪. Aim: to study the effects of thrombin and inflammatory factor IL-1 尾, TNF- 伪 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- 伪 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- 伪 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- 伪 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- 伪 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- 伪 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- 伪 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- 伪 10ng/ml (P0.05). Conclusion: (1) Thrombin and IL-1 尾 (TNF- 伪) 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- 伪 gradually increased in a time dependent manner. (2) thrombin acted alone with IL-1 尾 or TNF- 伪. The fibroblast release of MMP-9 was enhanced, but the combination of thrombin and IL-1 尾, TNF- 伪 decreased the release of MMP-9. (3) the combination of IL-1 尾 and TNF- 伪. There was no change in the release of MMP-9 from fibroblasts.
【学位授予单位】：汕头大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R751
本文编号：2302079
[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- 伪. Aim: to study the effects of thrombin and inflammatory factor IL-1 尾, TNF- 伪 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- 伪 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- 伪 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- 伪 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- 伪 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- 伪 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- 伪 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- 伪 10ng/ml (P0.05). Conclusion: (1) Thrombin and IL-1 尾 (TNF- 伪) 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- 伪 gradually increased in a time dependent manner. (2) thrombin acted alone with IL-1 尾 or TNF- 伪. The fibroblast release of MMP-9 was enhanced, but the combination of thrombin and IL-1 尾, TNF- 伪 decreased the release of MMP-9. (3) the combination of IL-1 尾 and TNF- 伪. There was no change in the release of MMP-9 from fibroblasts.
【学位授予单位】：汕头大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R751
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