Effects Of Angelica Sinensis Polysaccharide On Hepcidin Expression And Iron Overload

BackgroundAngelica sinensis polysaccharide(ASP)is the main active ingredient of Angelica,in particular,the role of the pharmacological effects of ASP in cancer research has made rapid progress.Studies have shown that iron in various cancer tissues was significantly higher than in normal tissue iron overload promotes tumor cell proliferation,metastasis and promote tumor angiogenesis,to iron therapy may be a new anti-tumor approach.Hepcidin is the key substances regulate body iron homeostasis is mainly synthesized and secreted by the liver cells,which can bind and degrade the cell surface membrane iron transporter protein(ferroportin,FPN),reducing the intracellular iron transport out,causing the cells to increase bioavailable iron.ObjectivesThis study intends to analyze ASP affects on the regulation of Hepcidin and iron metabolism in vivo,in order to provide a scientific basis for ASP clinical treatment of cancer.Methods1.Human normal liver L-O2 cells and hepatoma HepG2 cells treated with different concentrations of ASP,by reverse transcription-PCR(RT-PCR),Western blotting and other methods to observe the regulation of ASP on Hepcidin.2.By making iron overload animal models,Iron content of mice serum was detected by multi-function automatic biochemical analyzer.Iron content of mice liver,spleen and heart tissue were detect through the air-acetylene atomic absorption method and HE staining methods.Results1.Determine the optimal concentration and induction time of ASPRT-PCR results showed that: 100 mg / L ASP induced HepG2 cells(F = 329.938,P = 0.000)and L-O2 cells(F=411.918,P=0.000)after 48 h,Hepcidin mRNA were significantly upregulated;200 mg / L and 400 mg / L ASP induced HepG2 cells and L-O2 Hepcidin mRNA were significantly down-regulated and has a certain dose-dependent manner.Selection of 400 mg / L ASP induced HepG2(F = 168.292,P = 0.000)and L-O2 cells(F=35.631,P = 0.000)24 h,48 h,72 h and 96 h,RT-PCR results showed that: with the 400 mg / L ASP continuously,Hepcidin mRNA expression was significantly reduced and rendering the time-dependent.Given the above findings,the follow-up experiment use 400 mg / L ASP induced HepG2 cells and L-O2,time was 96 h.2.The effect of ASP on Hepcidin protein expressionWestern blot results showed that: with 400 mg / L ASP induced HepG2(t =24.825,P = 0.000)and L-O2 cells(t=54.157,P = 0.000)96h,significantly down-regulated expression of Hepcidin,the inhibition rate was 62% and 74%.3.produce iron overload miceCompared with the control group,the model group appears the second week of fur rough,dull,tired and slow growth;the third week mouse diet,activity was significantly reduced,nose,ears,tail color gradually deepened.Compared with the control group,serum iron high iron content model mice was significantly higher(t=29.321,P= 0.000),increased 2.50 times,The preparation of iron over-load model of C57BL/6 mice was successful.4.ASP increased serum iron content in C57 BL / 6 mouseserum iron increased significantly in model+ ASP group compared with model+ NS group,and it increased 1.36 times(t=10.362,P=0.000).5.the iron load of C57BL/6 mice tissue was reduced by ASP.Iron content in liver,heart and spleen was lower in model+ASP group than in model+ NS group(P=0.000)and they were reduced by 39.81%?38.73% and 36.84%,respectively.HE staining staining showed that positive site was brown particulate,the model + ASP group liver,spleen and heart iron were significantly lower than the model + NS group.Conclusions1.ASP could directly inhibit the expression of hepcidin.2.The iron load of C57BL/6 mice tissue was reduced by ASP.