Effects Of Grape Seed Proanthocyanidins On The Liver Function In Iron Overload Rats

[Objective]The study was to measur the liver bivalent mineral elements,hepatic metabolism related enzymes,the liver morphology,the expression of liver iron metabolism related genes,and to explore the function of grape seed procyanidins on liver function of iron overload rat.The results could provide some theoretical basis for the development of a natural iron reduction factor.[Methods]Choosing 200 ± 5 g SPF and 60±5g SPF male SD rats,we divided them into 4 groups randomly:control group,procyanidins group,iron load group and the experimental group,10 mice for each group.After adaptive feeding for 7d,100 mg/kg bw/day dose iron dextran every other day and 2.5 mg/kg bw/day dose iron dextran every day was intraperitoneal injected to build the acute and chronic iron overload rat model.The rats in experimental group injected 100 mg/kg bw/day dose iron dextran every other day and 2.5 mg/kg bw/day dose iron dextran every day,at the same time lavaged with 100 mg/kg bw/day and 25 mg/kg bw/day every day dose of procyanidins.Procyanidins group was lavaged with 100 mg/kg bw/day and 25 mg/kg bw/day every day dose of procyanidins.Using 0.9%of the normal saline to eliminate stress difference.The does was 1 mL/100 g bw every time.Tests time was 10 days and 7 weeks respectively.Taking the liver of rats and weighting the rat body and liver accuratly to calculate the viscera/body ratio after the feeding;exploring the content of Fe,Ca,Mg,Zn,Cu,Mn using ICP method,determinating the activity of ALT,AST,MDA,SOD,and content of-SH using the related kits;observing the changes of liver biopsy,laminin and cell apoptosis using HE staining,prussian blue staining,immunohistochemical staining and tunnel staining;measuring the the quantity of liver Hepcidin,Ferroportion,DMT1,TfRl,TfR2,Fas,Bcl-2 to using RT-PCR method.[Results]:(1)Compared with control group,the contents of Fe?Cu?Mn of acute procyanidins group decreased significantly or extremely significantly decreased(P<0.05 or P<0.01),the contents of Ca?Mg?zinc were no significant changes;the content of Ca in chronic procyanidins group extremely significantly increased(P<0.01),the content of Mn decreased significantly(P<0.05)and the contents of Fe,Mg,Zn,Cu were no significant changes.The contents of Fe,Mg,Zn,Mn in acute iron load group significantly increased or extremely significantly increased(P<0.05 or P<0.01),Cu content was significantly lower(P<0.05),the content of Ca had no significant change;the content of Fe,Ca,Zn,Mn of chronic iron load group extremely increased(P<0.01),there was no significant changes of the content of Mg,Cu.Compared with the load of iron group,the contents of Ca?Cu in acute experimental group were significantly lower(P<0.05).The zinc content significantly increased(P<0.05),the contents of Fe,Mg,Mn had no significant differences;the contents of Fe,Zn,Mn of chronic experimental group are extremely significantly lower(P<0.01),the contents of Ca,Mg,Cu had no significant differences.(2)Compared with control group,viscera index of acute procyanidins group extremely decreased(P<0.01),viscera index of chronic procyanidins group extremely increased(P<0.01).Acute iron load group has a rising trend,but the differences were not significant,viscera index of chronic iron load group significantly increased(P<0.01).Compared with iron load group,the viscera index of acute and chronic test group extremely decreased(P<0.01).(3)Compared with control group,the MDA content of acute procyanidins group is significantly lower(P<0.05),the level of ALT?AST?-SH?SOD had no significant differences;the level of ALT?-SH of chronicprocyanidins group had no significant differences,the AST?MDA?SOD contents of chronic procyanidins group were significantly lower(P<0.01).The activity of ALT?AST of acute iron load group significantly or extremely significantly increased(P<0.05 or P<0.01),the content of-SH decreased significantly(P<0.05),MDA and SOD content had no significant differences;the activity of MDA?-SH?SOD of chronic iron load group extremely significantly decreased(P<0.01),there wrer no significant differences in the activity of ALT?AST?Compared with the iron load group,the activity of ALT and AST of acute experimental group is significantly lower(P<0.05)and the activity of SOD extremely increased(P<0.01),the content of MDA,-SH had no significant differences;the activity of SOD in chronic experimental group significantly increased(P<0.05),the activity of ALT?AST?MDA?-SH had no significant differences.(4)Compared with control group,the rat liver cells of the procyanidins group was normal,the cells did not have iron deposition,the quantity of layer adhesion protein of acute procyanidins group have no significant differences,but the quantity of layer adhesion protein of chronic procyanidins group significantly decreased(P<0.05),the number of apoptotic cells had no significant differences;the liver cell structure of the iron load group was disorded,and it had structure disorder,pigment deposition,hemorrhage and inflammatory symptoms,the cell apoptosis is serious,the quantity of layer adhesion protein increased,significantly,and the quantity of layer adhesion protein of acute iron load group significantly increased(P<0.05).The apoptotic cells of acute iron load group increased,the apoptosis cells of chronic iron load group reduced.Compared with iron load group,experimental group's liver cell structure was also disorder,but the inflammation,pigment deposition reduced with slightly fat degeneration,the quantity of layer adhesion protein and the number of apoptotic cells was significantly lower or extremely significantly lower(P<0.05 or P<0.01),pathological changes was lighter than iron group as a whole.(5)Compared with the control group,the Hepcidin,DMT1,TfR2,Fas,Bcl-2 gene expression of acute Pas group significantly decreased or extremely significantly decreased(P<0.05 or P<0.01),the Ferroportin,TfRl gene expression did not change significantly.The Hepicdin,TfR2,Fas,Bcl-2 gene expression of chronic Pas group decreased significantly(P<0.01),the DMT1,TfRl gene expression increased extremely(P<0.01),the Ferroportin gene expression did not change significantly.Compared with the control group,the Hepicdin?DMT1?Bcl-2 gene of the acute iron overload group expression decreased significantly or extremely significantly decreased(P<0.05 or P<0.01),Ferroportin,TFR1,TFR2,Fas gene expression did not change significantly.The hepcidin gene expression of the chronic iron overload group extremely significantly decreased(P<0.01),The DMT1,TfR1,TfR2 gene expression of the chronic iron overload group extremely increased significantly(P<0.01),Ferroportin,Fas,Bcl-2 gene expression did not change significantly.Compared with the iron overload group,Ferroportin gene expression of acute experimental group decreased significantly(P<0.05),Bcl-2 gene expression increased significantly(P<0.05),Hepcidin,DMT1,TfRl,TfR2,Fas gene expression did not change significantly.Ferroportin,DMT1,TfR1,TfR2,Fas gene expression of chronic experimental group decreased significantly or extremely significantly decreased(P<0.05 or P<0.01),hepicdin,Bcl-2 gene expression did not change significantly.[Conclusions]:Procyanidins could potentially adjust the function of iron overload rat liver through regulating the liver bivalent mineral elements,hepatic metabolism related enzymes,the liver morphology and the expression of liver iron metabolism related genes.