激活Toll样受体3对皮肤成纤维细胞释放MMP和胶原蛋白的作用机制研究

It plays an important role in wound healing to activate nature immunity by stimulation with the ligand of Toll like receptor. There are many kinds of cells involving in the process of wound healing. It is also required that the proliferation,transformation and differentiation of fibroblast,collegen synthesis and matrix metalloproteinases play its role in wound healing. Previous study showed that there existed different kinds of Toll like recetors and the diffenent expression and activity of matrix metalloproteinases in fibroblast in skin injury.The underlying rationale of poly I:C therapy is that an smearing of poly I:C at the site of injury may promote tissue repair via cytokine or matrix metalloproteinases release from fibroblasts. The molecular mechanisms of poly I:C in the skin wound healing process are not well understood at present, and would benefit from clarification. .The goal of this proposal is to understand the mechanissms of poly I:C affecting the expression and activity of matrix metalloproteinases in fibroblast. .In this proposal, firstly we will investigate the status and dynamic of Toll like receptors in primary cultured human fibroblast, in the isolated mice fibroblast from pre or post injury, and from different time points after injury. The receptor will be detected by immunofluorecence, western blot and RT-PCR and q-PCR. Secondly, we will investigate the effect of the ligand of Toll like receptor on the proliferation, migration of fibroblast, the synthesis of collegen in fibroblast and the expression and activity of matrix metalloproteinases in fibroblast. Lastly, we will reveal the invovled of signal transduction pathway and transcription factors in regulation of the transcription of matrix metalloproteinases. .Our study will increase our knowledge of wound healing and help us to find a new strategy to enhance wound healing.

免疫激活在伤口愈合过程中起重要作用。我们分别在Toll样受体3（TLR3）基因敲除小鼠伤口模型和人皮肤激光伤口模型中用TLR3的配体poly I:C激活免疫，发现对伤口愈合起促进作用。前期的实验结果提示激活TLR3可增强鼠单核细胞分泌趋化因子，可促进成纤维细胞分泌金属蛋白酶9和2。我们推测通过TLR3激活免疫作用可调节皮肤成纤维细胞分泌MMPs和促进胶原合成，导致成纤维细胞生长和细胞群的迁徙。从而加速伤口愈合。而这一作用的分子调控机制目前仍然不清。本研究拟在永生化皮肤角朊细胞和原代培养的成纤维细胞上，在TLR3敲除小鼠模型和TICAM 小鼠（敲除了polyI:C处理对INF和TGF反应基因）模型上，分别在整体、细胞和分子水平层次上鉴定和探讨对参与的免疫活性分子，细胞信号转导通路和转录因子对相应MMPs和胶原合成影响机制。对免疫影响伤口愈合提供新的理论认识并有助于找寻新的靶点。

免疫激活在伤口愈合中起重要作用。我们分别在TOLL样受体3基因敲出鼠模型和人皮肤伤口模型中用TLR3配体PolyI:C激活免疫，发现对伤口愈合起促进作用。前期实验结果提示激活TLR3可增强鼠单核细胞分泌趋化因子，可促进成纤维细胞分泌金属蛋白酶9和2。我们推测通过TLR3激活免疫作用可调节皮肤呈纤维细胞分泌MMPs和促进胶原合成，导致呈纤维细胞生长和细胞迁徙。从而加速伤口愈合。而这一作用的分子调控机制目前仍然不清，本研究拟在元代培养的呈纤维细胞上，在TLR3敲除小鼠模型上，分别在整体、细胞和分子水平层次上鉴定和探讨对参与的免疫活性分子，细胞信号转到通路和转录因子对相应MMPs和胶原影响机制。对免疫影响伤口愈合提供新的理论认识并有助于找寻新的靶点。