Study On Pathological Metabolism Of Glycosaminoglycans In Vascular Smooth Muscle Cells By Phlegm Medicine Chenpi

ObjectiveAtherosclerosis(AS)is one of the cardiovascular diseases that endanger human health.The proliferation and transmembrane migration of smooth muscle cells in the media of the vascular wall plays an important role in the development of atherosclerosis.Modern studies have shown that the pathological metabolism of glycosaminoglycans(glycosaminoglycans,GAGs)in smooth muscle cells of the media has an important relationship with the accumulation of many inflammatory factors[1-3].The pathological metabolism of glycosaminoglycans in smooth muscle cells is closely related to traditional Chinese medicine phlegm syndromes,and the clinical effects of Huatan medicine on atherosclerosis therapy are significant[4-6].In this study,oxidized low_density lipoprotein(Ox_LDL)was used to make vascular smooth muscle cell proliferation model,to investigate the effect of phlegm medicine Chenpi on the pathological metabolism of glycosaminoglycans in vascular smooth muscle cells.MethodsThe vascular smooth muscle cell proliferation model was made by ox_LDL;the amount of total GAGs in vascular smooth muscle cells were detected by carbazole sulfate method;the changes of GAGs species in vascular smooth cells were detected by acetate membrane electrophoresis;the amount of total chondroitin sulfate(CS)in vascular smooth muscle cells were detected by high performance liquid chromatography;the fluorescence intensityof the fibrolast growth fatctors-GAGs-fibrolast growth receptors(bFGFs-GAGs-FGFRlc)were detected by flow cytometer;the changes of transcriptome expression in GAGs of vascular smooth muscle cells were detected by high-throughput sequencing.Results1.Effect of Chenpi on the total amount of GAGs in vascular smooth muscle cells.Compared with the blank control group,the total amount of GAGs in the model group increased(P<0.05).Compared with the model group,the total amount of GAGs in the 100?g·ml-1 Chenpi group decreased(P<0.01).2.Effect of Chenpi on the species of GAGs in vascular smooth muscle cells.Compared with the blank control group,the number of CS in the model increased(P<0.05);compared with the model group,the CS production in the 100 ?g·ml-1 Chenpi group decreased(P<0.01).3.Effect of Chenpi on the amount of galactosamine in vascular smooth muscle cells.Compared with the blank control group,the amount of galactosamine was increased in the model group(P<0.05).Compared with the model group,the amount of galactosamine was decreased in the 100 u g·ml-1 Chenpi group(P<0.01).4.Effects of Chenpi on the fluorescence intensity of the ternary complex of bFGF-GAGs-FGFRlc in vascular smooth muscle cells.Compared with the blank control group,the fluorescence signal of the model group was increased(P<0.05).Compared with the model group,the fluorescence signal of the 100?g·ml-1 Chenpi group was weakened(P<0.01).5.Effect of chenpi on transcriptome expression of GAGs in vascular smooth muscle cells.Compared with the blank control group,the fragment per kilobase of transcript per million fragments mapped(FPKM)of Cspg5 was increased in the model group(P<0.05);Compared with the model group,the Cspg5 FPKM value was decreased in the 100 ?g·ml-1 Chenpi group(P<0.01).ConclusionPhlegm medicine Chenpi may play a role in anti-atherosclerosis by decreasing the synthesis of chondroitin sulfate in vascular smooth muscle cells,decreasing the production of bFGF-GAGs-FGFRlc ternary complex,down-regulating the expression of Cspg5,and regulating the metabolism of glycosaminoglycans.