The Effect Of Usnic Acid On Tumor Angiogenesis And BFGFR/HSPG Mediated Signaling Pathway

Objective: This project through the usnic acid on tumor vessels and bFGFR1/HSPG to explore the influence of usnic acid on tumor angiogenesis for usnic anti tumor and clinical application,research and develop new drug of usnic acid inhibition of tumor angiogenesis provide experimental basis.Method: Usnic acid was selected as test substance,with the mouse H22 tumor model as the subjects,by HE staining of tumor tissue sections and immunohistochemistry detect pathological index MVD(microvessel density),ELISA method was used to detect the expression of VEGF and bFGF in the serum of mice.The expression of PCNA(proliferating cell nuclear antigen) was detected by blotting Western WB assay,RT-PCR detect bFGFR1,HSPG expression,to study the effect of test substance on tumor growth and angiogenesis in vivo experiment,and to explore its mechanism.By the method of CCK-8 to study the effects of different concentrations of usnic acid on the proliferation of HUVEC,preliminary determine the best usnic acid concentration; RT-PCR technology detect the expression of bFGFR1,HPSG mRNA of HUVEC and WB detect PCNA,bcl-2,Bax expression.Result: In vivo experiment,compared with the model group,the mice H22 tumor volume and of usnic acid group quality growth was significantly slower(P<0.05),HE staining showed that the pathological damage of usnic acid group the tumor tissues were significantly lower,and immunohistochemical results of usnic acid group showed that tumor microvessel density(relative positive region 160249±1256)was significantly reduced(P<0.05),usnic acid can significantly reduce serum VEGF and bFGF levels in mice(P<0.01),WB showed that the expression of PCNA protein of usnic acid group was significantly lower,RT-PCR showed that usnic acid group of bFGFR1,HSPG,gene expression was significantly lower.In vitro experiment,CCK-8 experiment showed that usnic acid can significantly inhibit the proliferation of HUVEC cells,usnic acid can promote HUVEC cell apoptosis,and usnic acid can significantly reduce the levels of bFGF and HUVEC based VEGF cell supernatant(P<0.01).WB showed that usnic acid group of PCNA,bcl-2,Bax protein expression wassignificantly lower,RT-PCR showed that usnic acid group of bFGFR1,HSPG gene expression was significantly lower.Conclusion: usnic acid can inhibit the H22 Tumor Growth and angiogenesis and inhibited the expression of VEGF and bFGF,inhibiting PCNA,Bax,bcl-2 protein expression,inhibiting bFGFR1,HSPG gene expression,inhibiting the proliferation of HUVEC in vitro suppressed tumor vessel formation.