Preliminary Study On The Effect Of Flavonoids On The Activity Of BFGF

In the process of growth and metastasis,tumor cells can promote angiogenesis by secreting various angiogenic factors,and then promote tumor metastasis.Basic fibroblast growth factor(bFGF)is an effective angiogenic factor.At present,the development of tumor drugs targeting bFGF has become a hot spot.Tetrastigma hemsleyanum Diels et Gilg is a unique and precious Chinese medicinal materials in China.A large number of studies have shown that T.hemsleyanum has anti-tumor activity.Therefore,using bFGF as target molecule to screen affinity factors in T.hemsleyanum that can interact with bFGF,and the effect of affinity factors on bFGF activity is the focus of this paper.In this study,bFGF affinity chromatography was used to screen and identify the specific binding components of bFGF in T.hemsleyanum.Affinity chromatography,UV-vis spectrophotometry,circular dichroism and non-denatured polyacrylamide gel electrophoresis were used to determine the affinity between FGFC and bFGF affinity factors in T.hemsleyanum.At the same time,mouse embryonic fibroblasts Balb/c 3T3 and neuroblastoma cell SH-SY5 Y were used to construct the models.MTT method and synaptic growth were used to evaluate the effect of affinity factors on the biological activity of FGFC.In order to improve the bioavailability and efficacy of catechin,acylation modification was used to study the synthesis of catechin derivatives.The results were as follows:1.The screening results of affinity factors of bFGF in Trifolium repens showed that they are catechins and procyanidin B1.2.Affinity chromatography showed that catechins and procyanidin B1 bound to FGFC,and no eluting substances were observed in PBS and glycine solutions.The results of UV-vis spectrophotometry showed that after catechin binding to protein,the absorption peak of peptide bond at204 nm shifted red.After binding with procyanidin B1,the absorption peak of peptide bond at 204 nm became stronger and red shift.Circular dichroism showed that both substances enhanced the positive and negative peaks of protein CD spectra.The results of non-denatured polyacrylamide gel electrophoresis showed that catechins and procyanidin B1 inhibited the migration of FGFC in a dose-dependent manner.All the above results showed that catechin and procyanidin B1 could bind to FGFC and changed the conformation of the protein.3.The results of Balb/c 3T3 cell model test showed that catechin of 1mg/mL significantly inhibited the proliferation of Balb/c 3T3 cells,and 500 ?g/mL and 1mg/ml catechins and procyanidinB1 significantly inhibited the proliferation of Balb/c 3T3 cells at 25 ng/mL FGFC.The inhibitory effect of catechin on FGFC was the strongest when the concentration was 1mg/mL,and the activity of procyanidin B1 was the strongest when the concentration was 500 ?g/mL.4.The results of SH-SY5 Y cell model test showed that catechin and procyanidin B1 had cytotoxicity above 50 ?g/mL.The concentration of catechin and procyanidin B1 significantly inhibited the activity of FGFC from 1 ?g/mL to 50 ?g/mL in a dose-dependent manner.5.The synthetic experiment of catechin derivatives shows that the preliminary synthesis of catechin derivatives proves the feasibility of acylation modification method.