家兔TIMP1基因的克隆表达及其对毛乳头细胞增殖的影响

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中图分类号：S829.1 文献标志码：A 文章编号：0366-6964（2025）10-4988-10

Abstract： The aim of this paper was to clone and analyse TIMPl，a member of the tissue inhibitor of matrix metalloproteinases（TIMPs） family. The regulatory role of TIMPl on hair follicle growth-related genes in hair follcle papilla cells （DPCs） was explored by overexpression and knockdown of TIMP1. In this study，we cloned the coding sequence（CDS） of TIMP1 and analysed its bioinformatics function. After that，we constructed the overexpression vector pcDNA3.1-TIMPl and designed and synthesized siRNA. We overexpressed and knocked down TIMP1 in DPCs to probe the expression of genes related to hair folicle growth and development， and detected the proliferation level of DPCs by EdU and CCK-8. The results showed that the CDS region of the rabbit TIMP1 gene was 624 bp in length，encoding a total of 207 amino acids. Bioinformatics analysis indicated the presence of a signal peptide in the TIMPl protein，which did not contain a transmembrane structural domain and was homologous in different mammals. Overexpression of TIMP1 in DPCs was able to highly significantly up-regulate the mRNA expression levels of BMP2，SFRP2，and TGFβ1 genes （ P<0.01 ），and highly significantly down-regulate the mRNA expression level of WNT2 gene （ P<0.01 ），and knockdown of TIMP1 was able to significantly down-regulate the expression of mRNA of BMP2， SFRP2 genes （ ⋅P<0. 05 ），and highly significantly down-regulated the mRNA expression of TGFβ1 gene （ P<0.01 ）.In addition，EdU and CCK-8 results showed that overexpression of TIMP1 could inhibit the proliferation of DPCs，and knockdown of TIMP1 could promote the proliferation of DPCs.In this study，we successfully cloned the CDS sequence of rabbit TIMP1 gene and preliminarily predicted its bioinformatics function，analysed the regulatory effect of TIMP1 on hair follicle growth-related genes. We verified its effect on inhibiting DPCs proliferation，and provided a reference for elucidating the theoretical research of rabbit hair folicle growth and development.

Keywords：TIMPl；hair follicle；domestic rabbit；dermal papilla cells ∗ Corresponding author： WU Xinsheng，E-mail： xswu@yzu. edu.cn

毛囊（hairfollicle，HF）是一种位于真皮层且具有复杂结构的微型器官，能够保护皮肤组织，并参与体温的调节[1，2]。（剩余15003字）