Initial Research On The Effect And Mechanism Of Tivozanib On Pulsed Dye Laser Induced Angiogenesis

Port wine stains(PWS)are a capillaries malformations,which appear more likely to occur on the head or face,seriously affecting the mental and spiritual health of patients.PWS can be used as a single disease or as a clinical manifestation of multiple syndromes.This paper presented a unique case of type IIb Phakomatosis pigmentovascularis(PPV)in association with Klippel-Trenaunay syndrome(KTS),scleral melanosis,and rare extensive PWS lesion,and PWS-related syndromes were also reviewed as a clue.PWS skin lesions are difficult to cure,especially those manifested large and progressively developed in the related syndromes.Pulsed dye laser(PDL)is the main method for the treatment of PWS,while clinical studies have shown that some patients still have low clearance rates and are resistant to treatment.Research has shown the reason for the poor effect of PDL treatment is PDL-induced angiogenesis after laser therapy.Vascular endothelial growth factor(VEGF)plays an important role in PDL-induced angiogenesis.It can activate the tyrosine kinase activity of vascular endothelial growth factor receptor(VEGFR),trigger a full range of responses in endothelial cells,and then participate in the regulation of angiogenesis.Tivozanib is a VEGFR tyrosine kinase activity inhibitor,which can block the angiogenesis effect of VEGF and reduce vascular permeability and has approved for the treatment of advanced and metastatic renal cell carcinoma on Clinical works.Recently research about Tivozanib concentrate mainly on various tumor diseases.This paper aims to discuss the effect and mechanism of Tivozanib on PDL-induced angiogenesis,and provide a new feasible plan for clinical treatment of PWS.In this study,different PDL energy densities were used to irradiate the abdominal skin of rats.According to the general and pathological changes of the irradiated area,8J/cm~2 was set for successive tests.Under this energy density,the skin surface had less scab and the blood vessel destruction effect was relatively strong.Divide the rat skin into 4 areas,irradiate 3 of them uniformly with an energy density of 8J/cm~2,and apply different concentrations of Tivozanib and its matrix components to the laser irradiation areas.Grouped as follows:(1)vacant group(without medicine,no laser irradiation),(2)control group(drug base,laser irradiation),(3)0.5%Tivozanib group,(4)1%Tivozanib group.Use camera and dermoscopy for gross observation,HE staining,immunohistochemical staining,blood vessel counting to detect vascular angiogenesis.The results showed that the 0.5%Tivozanib group and 1%Tivozanib group showed significant angiogenesis inhibitory effects than the control group on the7,10,and 14 days,and 1%Tivozanib group showed significant angiogenesis inhibitory effects than 0.5%Tivozanib group.The results indicate that Tivozanib successfully inhibited PDL-induced angiogenesis,and the inhibitory effect of 1%Tivozanib was more significant than that of 0.5%Tivozanib.In order to explore the mechanism of Tivozani's inhibition of PDL-induced angiogenesis,the 7th day when the general and pathological results were significantly different was selected as the time point,and the 1%Tivozani group with the strongest vascular inhibitory effect and the control group were selected as organize samples for transcriptome sequencing.The results of transcriptome sequencing revealed a total of588 significantly differentially expressed genes,including 90 up-regulated genes and498 down-regulated genes.GO enrichment analysis showed that 1004 functional items were significantly enriched during the whole action process(q<0.05).Differentially expressed genes showed higher enrichment items in regulation of response to external stimulus?immune system process?cell surface receptor signaling pathway?cell motility?cell migration?cell chemotaxis,etc.KEGG enrichment analysis showed that a total of 20 metabolic pathways were significantly enriched(q<0.05),in which cytokine-cytokine receptor interaction,PI3K-Akt signaling pathway,focal adhesion,chemokine signaling pathway,Ras signaling pathway,and RAP1 signaling pathway were the most important enrichment pathways related to angiogenesis.Finally,Real Time Polymerase Chain Reaction(real-time PCR)verified the genes with high expression differences in the top ranking and closely related to angiogenesis,namely Cxcl1,Cxcl2,Cxcl3,Cxcl6,Ccl3,Csf3,IL1?,i NOS,Mmp9,Mmp13,Plau,Ets1,Spp1,Nr4a1.The results are consistent with the trend of transcriptome sequencing results,which accessed the reliability of this study.This study explored the inhibitory effect of Tivozanib on PDL-induced angiogenesis,and provided a new idea for the treatment of clinical PWS.Transcriptome sequencing studied the mechanism of Tivozanib's inhibition of PDL-induced angiogenesis,providing reliable clues for future indepth research.