The Protective Effect Of Huoxue Huatan Compatibility Improving Hyperlipidemic Mouse Accompanied With Iron Dysmetabolism

Objective:1. To observe the level of hepcidin on the hyperlipidemia model formed by High-Fat-Diet, detecting the relationship between hepcidin and endothelial dysfunction、oxidative stress.2. To investigate the protective effects of Huoxue Huatan compatibility improve endothelial function partly via attenuating hepcidin related BMP/smad signal pathway in hyper-lipidemic model mice induced by High-Fat-Diet and explore its probablity mechanism involving in lipid profile, inferring hepcidin related pathway.Methods:1.8 weeks male fpn1 Tek-cre mice randomly divided into two groups, one group were common diet. The other were fed with High-Fat-Diet, mice were collected blood in sterile tubes and sacrificed by cervical dislocation, liver and aortic were removed at 8 weeks. Serum lipids parameters, MDA, SOD were measured with an automated analyzer. Hepcidin, NO, ET-1 and IL-6 levels were determined using an enzyme linked immunosorbent assay.2.8 weeks male fpn1 Tek-cre mice, expect for 8 mice as control group were fed with standard diet, others were given High-Fat-Diet for 6 weeks to generate the hyperlipidemia model. Established model mice were randomly divided into five groups, they were daily intragastrically administered with normal saline / Atorvastatin suspension /Danshen decoction / Gualou decoction / Danshen-Gualou decoction respectively, last for 2 weeks, 8 in per group. And then, mice were collected blood in sterile tubes and sacrificed by cervical dislocation, liver and aortic were removed. Serum lipids parameters, MDA, SOD were measured with an automated analyzer. Hepcidin, NO, ET-1, IL-6 levels were determined using an enzyme linked immunosorbent assay. Immunohistochemical method performed to analyse the liver expression position of BMP-6 and Western Blotting tested relative expression on phosphorylation of smad1/5/8 and stat3.Results:1. Compared with the NOR group, the concentration of TC, TG, LDL-C, hepcidin in serum MOD group has significantly increased(P <0.05), which is parallel to the content of ET-1, IL-6, MDA in serum. There are obvious severe iron deposit in HE morphology, all of those will accelerate hyperlipidemic. Meantime, NO, SOD content have diminish remarkably statistical difference(P <0.05).2. After administered with atorvastatin and Chinese herbs decoction, serum lipid level has been modulated to different degree compared with MOD group and all treatment group hepcidin content has been improving significantly. Treatment group Liver iron deposition has been ameliorated. As well as, the concentration of ET-1, IL-6, MDA in serum were alleviated. NO, SOD level more superior to MOD group. Moreover, all the herbs could inhibit the expression of BMP-6 and reduce the ratio of p-stat3, p-smad1/5/8 to GAPDH. When compared with any two groups, the effective compatibility of Danshen Gualou show its protective effects are relatively superior to Danshen group or Gualou group.Conclusion:1. High-fat-diet generated fpn1 Tek-cre mice with obvious iron deposit and endothelial dysfunction and oxidative stress. At the same time, hepcidin level and related pathway were abnormal.2. In summary, the huoxue-huatan compatibility exert the lipid profile modulated effects to protect hyperlipidemic accompany with iron dysmetabolism. All those effects contributed to improve the function of endothelial, alleviating inflammatory, inhibiting oxidative stress, concurrent with our findings of hepcidin and related signal protein.