Effect Of Rhizoma Drynariae Total Flavonoids On Rat Liver Hepcidin Gene Expression

Objective:To explore the curative effect of Rhizoma Drynariae total flavonoids(RDTF)on osteoporosis(OP)rats,and to explore the effect of RDTF on the gene expression of liver hepcidin(HEPC)by ELISA,RT-PCR and immunohistochemistry.Methods:A total of 60 adult SD female rats were randomly divided into 20 sham operation(SHAM)group and 40 ovariectomized(OVX)group.The ovariectomized ovaries were ovariectomized and the osteoporotic models were made.The SHAM group was only removed the same amount of adipose tissue.After 3 months of modeling,10 rats were randomly selected from the SHAM group and 10 rats from the OVX group were killed.After the success of verify OP test,the OP model rats were randomly divided into 3 groups and named the ovariectomized blank(OVX+B)group,the low dose group of traditional Chinese medicine(OVX+L),the high dose group of traditional Chinese medicine(OVX+H),and 10 rats in each group.Two Chinese medicine groups were given RDTF(OVX+L:100mg/kg/d,OVX+H:200mg/kg/d)orally,and the control groups received equal volume of distilled water orally.After 4 weeks,blood,liver and right side femur were taken to test by serology,genetics,bone tissue morphology and immunology:?detect bone mineral density;?detect the content of serum calcium,iron;?detect the serum hepcidin content by Elisa;?detect the expression of liver hepcidin mRNA by RT-PCR;?observe the middle femur sections by HE staining;?detect the expression of BMP-6 in liver by immunohistochemistry.Results:?Test BMD:SHAM group>OVX+H group>OVX+L group>OVX+B group.There was no significant difference between the SHAM group and the OVX+H group(p>0.05),Comparing every double groups of the others,the difference has statistical significance(P<0.05).?The serum calcium level of the three groups was lower than that of the SHAM group,the difference was statistically significant(P<0.05),and the difference between the three groups was not statistically significant(P>0.05);the serum iron level was same.?Serum HEPC content:SHAM group,OVX+L group was higher than that of OVX+B group and OVX+H group,the difference was statistically significant(P<0.05);the difference between SHAM group and OVX+L group was not statistically significant difference(P>0.05);the difference between OVX+B group and OVX+H group was not statistically significant(P>0.05).?The content of liver HEPC mRNA:OVX+H group>SHAM group>OVX+L group>OVX+B group.SHAM group,OVX+L group and OVX+H group had statistically significant difference compared with OVX+B group(P<0.05);OVX+L group and OVX+H group had no significant difference compared with SHAM group(P>0.05);there was a significant difference between OVX+L group and OVX+H group(P>0.05).?Observation of HE sections by lightmicroscopic:bone plate density:SHAM group>OVX+H group>OVX+L group>OVX+B group;bone sag size:OVX+B group>OVX+L group>OVX+H group>SHAM group;Osteocyte number:SHAM group>OVX+H group>OVX+L group>OVX+B group;cortical surface roughness:SHAM group>OVX+H group>OVX+L group>OVX+B group.?Liver BMP-6 immunohistochemical results:the positive rate of BMP-6 expression:OVX+H group>SHAM group>OVX+L group>OVX+B group.There was no significant difference between the SHAM group and the OVX+H group(p>0.05).Comparing every double groups of the others,the difference has statistical significance(P<0.05).Conclusion:?Rhizoma Drynariae total flavonoids can improve rats osteoporosis,and the dose-response relation-ship is proportional to.?The ovariectomized method can reduce the contents of serum calcium and iron,but Rhizoma Drynariae total flavonoids can not affect serum calcium and iron levels.?Rhizoma Drynariae total flavonoids have influence on the content of hepcidin in serum of rats,which promote the expression of serum hepcidin in low concentration,the effect is not significant when the concentration is high.?OVX can inhibit the expression of HEPC in the liver,Rhizoma Drynariae total flavonoids can upregulate the expression of liver HEPC,and the dose-response relationship is proporti-onal to.?Rhizoma Drynariae total flavonoids can upregulate the expression of liver BMP-6,and the dose-response relationship is proportional to.