FAK通路对1型单纯疱疹病毒诱导的角膜上皮细胞MMP-2,9分泌、激活的调控机制研究

Focal adhesion kinase（FAK）is a non-receptor tyrosine kinase that participates in the process of cytobiology through multiple sites and pathways, with a comprehensive research prospect.In past studies, the applicant found that the expression and activity of matrix metalloproteinase (MMP)-2,9 increased significantly in the herpes simplex virus keratitis (HSK) ;Type 1 herpes simplex virus (HSV-1) can contribute directly to the secretion and activation of MMP-2 in primary cultured rabbit epithelial cells; TNF-α stimulates MMP-2 and MMP-9 activities via the activation of FAK/ERK signaling in human corneal epithelial cells.These results suggest that HSV-1 infection of the cornea may cause the secretion and activation of MMPs in the corneal tissue by activating FAK and its downstream members directly .The project intends to confirm the focal adhesion kinase (FAK) pathway's regulatory role to the secretion and activation of MMP-2 induced by HSV-1 through the application of RNA interference technology and gene gun corneal injection technology in cultured human corneal epithelial cell line and mouse HSK model,in order to provide a theoretical basis for seeking new HSK treatment programs.

粘着斑激酶（FAK）作为一种非受体蛋白酪氨酸激酶通过多位点多通路广泛参与细胞生物学过程，具有广泛的研究前景。申请人在过去的研究中发现基质金属蛋白酶(MMP)-2,9在单纯疱疹病毒性角膜炎（HSK）中的表达和活性明显增加；1型单纯疱疹病毒（HSV-1）可直接刺激原代培养的兔上皮细胞的MMP-2,9分泌和激活；肿瘤坏死因子(TNF)-α通过粘着斑激酶/细胞外信号调节激酶(FAK/ERK)通路增加MMP-2,9表达及分泌。上述结果提示HSV-1感染角膜后可能通过直接激活FAK及其下游成员导致角膜组织中MMPs的分泌和激活。本项目拟在体外培养的人角膜上皮细胞系以及小鼠HSK模型中应用RNA干扰技术及基因枪角膜注射技术证实粘着斑激酶（FAK）通路对HSV-1诱导的MMP-2,9分泌和激活的调控作用，为寻求新的HSK治疗方案提供理论依据。

粘着斑激酶是一种非受体蛋白酪氨酸激酶，可通过多位点多通路广泛参与细胞生物学过程。有研究发现HSV-1感染角膜后可能通过直接激活FAK及其下游成员导致角膜组织中MMPs的分泌和激活。本项目拟在体外培养的人角膜上皮细胞系以及小鼠HSK模型中应用RNA干扰技术及基因枪角膜注射技术证实粘着斑激酶通路对HSV-1诱导的MMP-2,9分泌和激活的调控作用。研究期间本项目通过体内及体外实验首次证实FAK/PI3K/Akt信号通路对HSV-1诱导的MMP-2,9分泌和激活具有调控作用，为HSK的发病机制提供了思路。此外，研究期间还发现Genistein可下调FAK/Src/JNK蛋白及其磷酸化蛋白表达水平，为HSK的临床治疗提供了新依据。