Objective: To investigate the effect of local autologous dermal fibroblasts transplantation on hypertrophic scar formation and wound healing quality in early scar formation. To explore the feasibility of fibroblasts for prevention and treatment of hypertrophic scar. Methods: Dermal fibroblasts were isolated from the dorsal skin tissue of New Zealand white rabbits by mechanical method combined with enzyme digestion. Passage 3 cells were induced to differentiate into osteoblasts and adipocytes. The complete epithelialization time and hypertrophic scar formation after full-thickness skin defect were confirmed by pre-experiment study. In the experiment, 6 rabbits were used, left ear as experimental group and right ear as control group. In the experimental group, the passage 4 dermal fibroblasts labeled with 5-bromodeoxyuridine (5-BrdU) were injected subcutaneously around the wound and hypertrophic scar on 20 d (day 2 after epithelialization) and 30 d (most obvious scar hyperplasia) after surgery. As a control group, physiological saline was injected following the same protocol. On 37 days after surgery, the hypertrophic scar tissue were harvested and assessed by gross view and histological examination. The transplanted cells were detected by immunofluorescence staining, transforming growth factor beta 1 (TGF-β1) and decorin(DCN) mRNA expression were assayed by real-time fluorescence polymerase chain reaction (RT-PCR), and the protein expression of TGF-β1、DCN、Collagen type Ⅰ and type Ⅲ were tested by enzyme linked immunosorbent assay(ELISA). Results: Compared with the control group, the scar in the experimental group was flatter and softer, the color was slightly lighter, and the volume was reduced. The histological results showed that compared with the control group, the number of fibroblasts and inflammatory cells in the superficial dermis was reduced, the proliferation of connective tissue and collagen deposition were reduced, and the basal cells and collagen fibers were arranged in order in the experimental group. The results of RT-PCR showed that TGF-β1 mRNA expression level in the hypertrophic scar tissue reduced significantly and DCN increased significantly in the experimental group, compared with the control group (P<0.01). ELISA results showed that TGF-β1, type Ⅰ and Ⅲ collagen contents in hypertrophic scar tissue were significantly lower than that in the control group, while the DCN expression was significantly increased (P<0.01). Conclusions The rabbit dorsal dermal fibroblasts significantly inhibited the hypertrophic scar in rabbit ears. The mechanism may be related to the up-regulation of DCN expression and down-regulation of TGF-β1 expression, reduction of type Ⅰ and Ⅲ collagen synthesis and extracellular matrix deposition, in the hypertrophic scar tissue.

AIM:To predict the mechanism of Sanguis draconis flavones(SDF)in the prevention and treat-ment of myocardial ischemia reperfusion injury(MIRI)based on network pharmacology and molecular docking methods.METHODS:The main chemical constituents of SDF were collected through literature search,and the targets of key con-stituents were screened by using the SwissTargetPrediction and TargetNet databases.Disease targets were also screened based on GeneCards,OMIM,TTD and PharmGkb databases,then targets were intersected with Cytoscape to construct the"drug-key constituent-target"network diagram,and the core target was obtained through visualization and analysis by Cytoscape software.Gene Ontology(GO)functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were analyzed by the Metascape platform.By utilizing AutoDock Vina software and Pymol,molecular docking between core compounds and core targets was carried out.Further,animal experiments were performed to explore the pharmacodynamic mechanism of SDF.RESULTS:The active constituents of SDF included loureirin B and loureirin A,which were mapped to 391 targets.A total of 3 096 MIRI disease targets were obtained from the database,af-ter intersection,172 intersection targets were obtained,and 56 core targets were acquired through analysis.The core relat-ed pathways included the cancer pathway and cell death signaling pathway.The results of molecular docking verified the strong binding activity between key constituents and key targets.Animal experiments demonstrated that SDF effectively prevented and treated MIRI,significantly inhibited the arachidonic acid 15-lipoxygenase(ALOX15)mRNA and protein expression,and reduced the myocardial infarction size after MIRI.CONCLUSION:SDF may play a positive role in the treatment of MIRI,which may be related to the regulation of the ALOX15 factor.