TLR7对瘢痕疙瘩形成中Smad信号通路的调控作用研究

发布时间：2018-05-08 18:38

本文选题：瘢痕疙瘩 + Toll样受体　；参考：《浙江大学》2016年硕士论文

【摘要】：目的研究Smads与TLR7的相关性,探讨TLR7对癜痕疙瘩形成中Smad信号通路的调控作用,以及对瘢痕疙瘩形成的影响。方法体外培养人成纤维细胞,使用TLR7激动剂Gardiquimod激动人成纤维细胞TLR7受体后,采用Western Blot方法检测Smad通路主要相关蛋白,即Smad4、7蛋白表达变化情况。通过向裸鼠皮下移植瘢痕疙瘩组织构建瘢痕疙瘩裸鼠模型,在动物体内实验水平,使用TLR7激动剂Gardiquimod皮下注射处理后,采用Western Blot方法检测Smad4,7蛋白表达变化情况,Western Blot各组凝胶印迹成像相对灰度值采用单因素方差分析、LSD-t检验、SNK-q检验,P0.05为差异具有统计学意义。对移植物处理后样本进行组织学观察,切片染色主要采用苏木精-伊红染色法(HE)染色和Masson染色。结果与对照组相比,人成纤维细胞体外使用TLR7激动处理后,Smad4、7蛋白表达未见明显改变。与对照组相比,在瘢痕疙瘩裸鼠模型中使用TLR7激动处理后,瘢痕疙瘩内Smad4蛋白表达未见明显改变；但是Smad7蛋白表达明显增加,差异具有统计学意义(P0.05)；组织学观察显示：与对照组相比,瘢痕疙瘩裸鼠模型使用TLR7激动处理抑制了瘢痕疙瘩组织在裸鼠体内的萎缩。结论在体外细胞培养中,激动成纤维细胞TLR7并不直接影响Smad4、7蛋白的表达；在瘢痕疙瘩裸鼠模型中,激动TIR7不影响瘢痕疙瘩Smad4蛋白表达,但可上调瘢痕疙瘩Smad7的表达,同时对瘢痕疙瘩的萎缩具有抑制作用。
[Abstract]:Objective to study the relationship between Smads and TLR7, and to investigate the regulation of Smad signal pathway in the formation of keloid and the effect of TLR7 on keloid formation. Methods Human fibroblasts were cultured in vitro. Human fibroblast TLR7 receptor was stimulated by TLR7 agonist Gardiquimod. The expression of Smad4P7 protein was detected by Western Blot method. Keloid model of nude mice was established by subcutaneous transplantation of keloid into nude mice. The experimental level of keloid was treated by subcutaneous injection of TLR7 agonist Gardiquimod in vivo. The expression of Smad4T7 protein was detected by Western Blot method. There was significant difference in relative grayscale value of gel blotting imaging in each group with single factor analysis of variance (ANOVA) and SNK-q test (P0.05). Histopathological observation was carried out on the grafts. The sections were stained mainly by hematoxylin-eosin staining and Masson staining. Results compared with the control group, the expression of Smad4 F7 protein in human fibroblasts treated with TLR7 in vitro did not change significantly. Compared with the control group, the expression of Smad4 protein in keloid was not significantly changed after TLR7 stimulation, but the expression of Smad7 protein was increased significantly in keloid nude mice. The histological observation showed that compared with the control group, keloid nude mice model treated with TLR7 inhibited the atrophy of keloid tissue in nude mice. Conclusion in vitro cell culture, TLR7 does not directly affect the expression of Smad4F7 protein in keloid nude mice, but agitated TIR7 does not affect the expression of keloid Smad4 protein, but can up-regulate the expression of keloid Smad7 in keloid nude mice. At the same time, it can inhibit the atrophy of keloid.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R622

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