The effect of Poria cocos polysaccharide on wound healing in diabetic rats analyzed based on RNA-sequencing

PENG Zhenxue; ZHOU Zhisen; LIU Canyu; ZHANG Xuanfen

doi:10.3969/j.issn.1673-7040.2024.07.012

Abstract：Objective To analyze the effect of poria cocos polysaccharide(PCP)on wound healing in diabetic rats based on RNA-seq.Methods From September to October 2023,24 SPF SD male rats were randomly divided into diabetic control(DC)group,normal control(NC)group and PCP group(8 rats in each group)in Department of Plastic Surgery,The Second Hospital of Lanzhou Univer-sitv.Previous experiments showed that 1％PCP aqueous solution could promote wound healing in diabetic rats.Wounds in PCP group were treated with 1％PCP aqueous solution on wet compress after operation,while wounds in normal control group and diabetic group were treated with normal saline wet compress.On the 7th day after operation,5 rats were randomly selected from each group to calculate the wound healing rate,and the wound tissue was cut and stained with HE to observe the pathological changes of the wound tissue in each group.RNA-seq was performed to screen out the significant differential genes in NC,DC group,and DC,PCP group,kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis,gene ontology(Ontology)analysis were performed(GO)enrichment analysis and Gene set enrichment analysis(GSEA)were performed.The remaining 3 rats in each group were photographed and recorded on the 14th day after op-eration,the wound healing rate was calculated,HE and Masson staining was performed.Results On the 7th and 14th day after injury,the wound of rats in each group was gradually reduced.The wound healing of NC group was the best,DC group was the worst,and PCP group was between DC group and NC group.HE staining showed that there were a large number of inflammatory cells infiltration in DC group and a small number of inflammatory cells infiltration in NC group at the same time point.The inflammatory cells in PCP group were between NC group and DC group.KEGG and GO enrichment analysis showed that the significantly different genes between DC group and NC group were mainly enriched in cAMP,cGMP-PKG and other signaling pathways.It is involved in biological processes such as keratinization,ep-ithelial development,and inflammatory response.The significantly different genes between PCP group and DC group were mainly enriched in PI3K-Akt,Calcium and other signaling pathways.It is involved in the regulation of angiogenesis,epithelial development,and regulation of endothelial cell proliferation and other biological processes.GSEA analysis showed that:The minor differential genes in DC and NC groups were enriched in interleukin-17(IL-17),nuclear factor nuclear factor kappa-B(NF-κB),Calcium and tumor necrosis factor(TNF)signaling pathways.DC and PCP groups were enriched in IL-17 and cGMP-PKG signaling pathways.Conclusion High glucose may in-terfere with wound healing in rats through cGMP-PKG signaling pathway;PCP may promote wound healing in diabetic rats by affecting PI3K-Akt and Calcium signaling pathways.