Overexpression Of Sox18 Was Used To Explore The Mechanism Of Transformation Of Mouse Dermal Fibroblasts Towards Papillary Cells Of True Fur

Sox18 is an important transcription factor,and Sox18 deficient individuals suffer from hypotrichosis-lymphoedema-capillary dilation syndrome(HLT),and also plays a regulatory role in the HH and Wnt signalling pathways.Sox18 and Sox2 can act together on dermal papilla cells,but the mechanism of Sox18 action in dermal papilla cells still needs to be further investigated.In this thesis,we propose to investigate the effect of altering Sox18 gene expression on the transformation of dermal fibroblasts into epidermal hair papilla-like cells,with the following major studies.The main research contents are as follows:1)The Sox18 gene was amplified to construct a Sox18 gene overexpression reporter gene vector,and the overexpressed Sox18 vector was introduced into c57 newborn mouse dermal fibroblasts by lentiviral transfection.The expression levels of dermal papilla cell-specific marker gene and specific marker protein were detected in dermal fibroblasts by qRT-PCR,cell immunostaining and alkaline phosphatase staining.2)Design Sox18 interference sequence,synthesize Oligo,construct Sox18 knockdown vector,introduce Sox18 knockdown vector into dermal papilla cells,determine the knockdown effect by qRT-PCR,cell immunostaining,detect the expression levels of dermal papilla cell related specific marker genes and specific marker proteins by qRT-PCR,immunofluorescence staining,alkaline phosphatase staining,etc.qRT-PCR,immunofluorescence staining,alkaline phosphatase staining and other methods were used to detect the expression levels of dermal papilla cell-related marker genes and proteins.3)Primary dermal fibroblasts,dermal fibroblasts overexpressing Sox18 and null-loaded dermal fibroblasts were combined with epidermal cells and injected into the subcutaneous and intradermal dermis of nude mice respectively to establish a subcutaneous and intradermal animal model of nude mice.This study verified that overexpression of Sox18 gene in c57 mouse dermal fibroblasts resulted in a significant increase in the expression of signature genes and proteins associated with dermal papillae.In the constructed animal model of hair follicle regeneration,the skin tissue of nude mice overexpressing Sox18 was sectioned and showed obvious thickening of blood vessels and thickening of skin tissue.The results showed that the overexpression of Sox18 gene promoted the transformation of dermal fibroblasts into hair papilla-like cells and to a certain extent promoted hair follicle regeneration.