| Background:Oral lichen planus(OLP)is one of the most common chronic inflammatory diseases in the oral mucosa.If OLP persists and undergoes repeated erosions,there is a 0.1%-4%probability of malignant transformation,so it is classified as a precancerous state by WHO.In OLP,the expression of vascular endothelial growth factor,intercellular adhesion factor and vascular cell adhesion molecule are significantly up-regulated,and inhibiting angiogenesis can effectively promote OLP healing,so abnormal angiogenesis plays an important role in the pathogenesis of OLP.It has proven that there are a lot of OLP-associated myofibroblasts(OLP/MFs)expressing α-smooth muscle actin in the OLP matrix,and the proliferation and migration ability of OLP/MFs are enhanced,and they can also generate angiogenesis-related growth factors,promote the migration and differentiation of vascular endothelial cells,so we proposed that OLP/MFs played a key role in OLP angiogenesis.However,the specific mechanism for OLP/MFs to regulate angiogenesis is still unclear.Galectin-3(Gal-3)can be expressed in fibroblasts,bind to vascular endothelial cells,promote angiogenesis,and induce abnormal blood vessels in a variety of inflammatory diseases and tumors.However,there is no report on the related research of Gal-3 and OLP angiogenesis.Objective:Study the expression of Gal-3 in OLP tissues and OLP/MFs,and verify whether Gal-3 from OLP/MFs promotes the migration and tube formation of HUVECs.Methods:The expression of Gal-3 in normal oral mucosa tissues and OLP tissues were detected by immunohistochemistry.CD34 was detected to analyze Microvessel density(MVD),and the correlation between the two indicators was analyzed.Normal fibroblasts(NFs)and OLP/MFs were isolated and cultured.Transcriptome sequencing was used to detect whether the Gal-3 gene was abnormally expressed in OLP/MFs and to analyze the interaction between Gal-3 and angiogenesis-related proteins.The expression of Gal-3 in the cells was detected by RT-PCR,Western blot,and ELISA.Finally,the Gal-3 overexpression lentivirus vector was constructed and transfected into OLP/MFs,and co-cultured with HUVECs,and performed Transwell migration experiment and tube formation experiment to verify the influence of Gal-3 on the biological characteristics of HUVECsResults:(1)Immunohistochemical revealed that the expression of Gal-3 in OLP tissues was significantly higher than that in normal oral mucosa tissues.The expression of Gal-3 and MVD both changed with the clinical classification,showing a positive correlation trend.Correlation analysis showed that MVD increased with the increase of Gal-3,and the correlation index r value is 0.8537.(2)OLP/MFs and NFs were successfully isolated and cultured.The transcriptome sequencing results showed that the expression of Gal-3 in OLP/MFs was abnormal and down-regulated compared with NFs.Screening through STRING database proved that Gal-3 was highly correlated with OLP angiogenesis,as its important regulatory molecule.RT-PCR verified that the Gal-3 mRNA expression in OLP/MFs was lower than that in NFs.WB results showed that the expression of Gal-3 in OLP/MFs was higher than that in NFs.ELISA detected a trace amount of Gal-3 in the supernatant of OLP/MFs,but it was also higher than that in NFs group.(3)The Gal-3 overexpression lentiviral vector was constructed,and successfully transfected into OLP/MFs,and co-cultured with HUVECs.Transwell migration experiment showed that in the Gal-3 overexpression group,more HUVECs migrated than the blank control group.The tube formation experiment showed that Gal-3 overexpression group constituted more tubes than the blank control group.The number of junctions,branch numbers and branch lengths of the HUVECs in the Gal-3 overexpression group were statistically higher than the blank control group.Conclusion:The up-regulation of Gal-3 in OLP was positively correlated with MVD,and Gal-3 from OLP/MFs promoted the migration and tube formation of HUVECs. |
PDF Full Text Request