| Objective:Fu-qi granule was preparation though the optimization, process and prescription screening, It is based on the guidance of Chinese medicine theory and base on the clinical experience "Fubai formula" in 302 hospital of people’s Liberation Army. The prescription is consists of astragalus root, Poria, Angelica sinensis, Fructus broussonetiae, donkey hide gelatin and Fer mented Cordyceps powder medicine composition, it has initially identified the granule formulation has certain effect of anti-hepatic fibrosis of hypoproteinemia. We were observated the treatment of liver fibrosis effect of Fu-qi granule and to explore the molecular mechanism of action. It was expectations to find the targets of anti-liver fibrosis of hypoalbuminemia with Fu-qi granule, to provide scientific experimental basis and reasonable treatment theory for clinical treatment of hepatic fibrosis.Methods:①The study on the preparation of Fu-qi granules:Adoptting modern preparation technology, combined with the decoction of traditional drug habits to choose suitable extraction, concentration and drying conditions, and to investigate the preparation of the granules and the molding technology process. Ultimately, it will be available for quality control and pharmacodynamics research of fu-qi granules.②The quality control of Fu-qi granules:To identification the effective composition and the content of main drugs by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) methods so that to ensure the quality of fu-qi granule is stable.③Research of the phaemacodynamics and mechanism of fu-qi granule:Through the establishment of CCl4-induced the hypoalbuminemia of liver fibrosis in rat or mice model and TAA-induced rat liver fibrosis model, to investigate fu-qi granule on the therapeutic effect of hepatic fibrosis hypoalbuminemia and protection effect on the BCG+LPS induced immunological liver injury in mice model. At the same time, the reduced with the mice spleen lymphocyte proliferation response is mainly induced by intraperitoneal injection cyclophosphamide, to study the effect of fu-qi granule on the immune function of mice.Results:①Based on the theory of traditional Chinese medicine, to determine the formulation of fu-qi granule, astragalus membranaceus, poria cocos, angelica, disappear real mixed with 8 times the amount of water decoction 2 times. The first time decoction, which time was 1.5 hours and the second decoction time for 1 hour. The extract obtained before it will be dry and to enrichment powder. Sieving powder and to blending, it made from cordyceps sinensis and dextrin granulating.②Fu-qi granule is through the analysis of the thin layer chromatography, the results show that real spots exist in a similar position of the thin layer plate and control medicinal material of angelica sinensis consistent spots. High performance liquid chromatography (HPLC) analysis of astragalus armour glycosides, which as the main index was determinationed and study of methodology. The results of precision, repeatability and recovery experiment and the relative standard deviation (RSD) were less than 3%. Sample specificity is showed that chromatographic peak of astragalus armour glycosides peak and other components to baseline separation of chromatographic peaks, there is no interference on the determination in the negative control solution.③Liver function detection:analysis of biochemical indexes is showed that Fu-qi granule can decrease the CCl4-induced hepatic fibrosis in mice or rats, and TAA-induced rat liver fibrosis. It leads to the increase of level of serum ALT, AST, ALP. However, reducing the level of TP, ALB and difference was significant (P< 0.05). Each dose group can be reduced serum PC-III, IV C, LN and HA level in different degrees of different factors induced liver fibrosis model animal, it have showed significant difference (P< 0.05). Serum ALT, AST, ALP content were increased and the level of TP, ALB were reduced in BCG+LPS induced immunological liver injury in mice, there were significant differences (P<0.05). Meanwhile, Fu-qi granule can improve the cyclophosphamide induced mice spleen lymphocyte proliferation responses. Hepatic histopathological examination:Different chemical factors inducing animal liver tissue model group were emerged fibrous tissue hyperplasia and forming hepatic lobule. Give to fu-qi granule after different doses, liver tissue pathology revealed liver fibrosis degree improved significantly.④A preliminary study on the mechanism of effect of FQG:a-SMA immunohistochemical positive test results show that positive cytoplasm appeared Brown and negative piece without coloring. Western blotting results show that p-mTOR and HIF-1α protein expression was reduced in the liver fibrosis model group, and there was obvious difference (p<0.05). Compared with model group, Giving fu-qi granule, the content of p-mTOR and HIF-1 alpha expression was increased, they have show that significant difference (p<0.05). TIMP-1 and MMP-9 of RT-PCR analysis showed that the expression of TIMP-1 was significantly increased and protein expression of MMP-9 was significantly reduced in CCl4-induced liver tissue in rat liver fibrosis model group, which showed that significant difference (P<0.05). In contrast, TIMP-1 protein expression is significantly reduced and remarkable increase of MMP-9 protein expression in the fu-qi granule group, and it was significant difference (P< 0.05).Conclusion:①Fu-qi granule preparation prescription medicine was selection through clinical application, to determine the preparation method, drugs including that astragalus membranaceus, poria cocos, angelica and broussonetia papyrifera, combined with modern preparation technology, The powder was mixed with Equus asinus L and Fermentative cordycepic fungal powder and the mixture was added dextrin to the preparation of FQG.②TLC method was used to identify the preparation of angelica and broussonetia papyrifera, astragalus glycosides content of FQG were determined by HPLC method to implementation the quality control of fu-qi granule.③ Pharmacodynamics and mechanism study of fu-qi granule showed that FQG can significantly treatment of hypoalbuminemia of liver fibrosis caused by various factors, its mechanism may be related to regulation of the mTOR/p70S6K signaling pathway. |
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