USP4介导的TGFβ信号转导通路对人增生性瘢痕成纤维细胞增殖的影响-外科学专业论文.docxVIP

PAGE PAGE IV 摘要 关键词：增生性瘢痕；USP4；TGF-β信号通路；Vialinin A； Ab Abstract PAGE PAGE V ABSTRACT Objective： To investigate USP4 in the important role of hyperplastic scar formation, especially in the regulation of TGF-β/Smad signaling pathway, to ensure the correlation of USP4, TβRI and Smad7 and elucidate the mechanism of hypertrophic scar by inhibiting USP4. It will provide important theoretical basis to cure hyperplastic scar as a drug target for USP4. Methods： Collecting 15 specimens of HS and normal skin tissue in HS patients from our plastic surgery puts in low temperature, which is quickly taken back to the lab (six months after injury). A part of tissue is washed by the saline with 4% neutral formaldehyde fixed and is dehydrated conventionally, embedded in paraffin and immunohistochemical staining; Western blot for detecting the different expression level of USP4 and TβRI in HS tissue and normal skin tissue. Hypertrophic scar fibroblasts (HSFB) and normal skin fibroblasts (NSFB) were cultured in vitro by using tissue combined with modified trypsin digestion with HS tissue and normal skin tissue. The third to the fifth generation of HSFB and NSFB in logarithmic growth phase were selected in the experiment. (1)take the fourth generation of FB, Western blot to detect USP4 and TβRI and Smad7 protein expression level of hyperplastic scar fibroblasts and normal skin fibroblasts; (2) Hyperplastic scar fibroblasts were cultured in cell culture plate for 24 hours. After that, HSFB were intervened by 5μmol/L Vialinin A which was the inhibitor of USP4 for 0 hours, 12 hours, 24 hours, 48 hours, then collecting cells respectively and the expression of USP4 and TβRI and Smad7 protein was detected with Western blot again; (3) HSFB whose density is 1×104 per hole was vaccinated in 96-well plate to culture for 24h, then they were intervened by 5μmol/L Vialinin and The MTT method was to testify the proliferation of HSFB and The cells unintervened were regarded as the control grou