The Effect Of Total Flavonoids Of Psoralea And Its Separated Components On Osteoblast And The Mechanism

Psoraleae is the dried fruit of Psoralea corylifolia L.which belongs to leguminous plant. It is pungent and warm nature drug.to warm and recuperate the kidney yang.Clinically,Psoraleae was used principally to treat diarrheaing due to Kidney Deficiency, frequent urination,impotenee,cold pain at waist and knee and coughing duetoasthenia-cold and so on. The modern studies indicate that Psoraleae can distend blood vessel,raise the myocardial contractile force,antitumor,antisepticise and so on. As yet, the professionals have got about forty compounds of coumarins,flavonoid and terpene from Psoraleae. The coumarins and flavonoid are important ingredients which can display the pharmaco-acticity.There are many kinds of flavone separated from Psoraleae,including osajin, flavonone, chalcone,and so on. It is indicated by the research that the total flavonoids of psoraleae not only possesses the effect of antioxygen and female hormone,but also prevent bone resorpted,and increase bone density,so the flavone plays an important role in the treatment on osteoporosis. The total flavone contains too many active ingredients to carry out the furth reseach on specific pharmaco-activity of each active compound. So, we will separate and purify the total flovone,and then, we can get the single component and study on the pharmacologic action to sieve the component for the clinical value and establish fundament to develop the new drugs.Firstly,it was the first time that the total flovone of Psoraleae was separated by the High Speed Countercurrent Chromatography（HSCCC）,and we established the separation technology initially. In the experiment,the solvent system was applied which was composed with N-hexane,acetic ether, methanol and water in proportion of 1:1:1:1. After demixing,the upper phase was collected as stationary phase and the bottom phase was collected as mobile phase,the temperature of separation was 25℃and the rotation speed was 800r/min,eventually,we got 6 peaks by detecting with 254nm ultraviolet, each peak represented one component from the total flovone.Then,the chemical property of 6 components were identified initially, we affirmed that the component of peak 4 and 5 are flavonol and flavonone of Psoraleae.Secondly,the expriment on osteoblast was carried out to detect the effect of the total flovone, the flovone and the flavonone on the proliferation and the differentiated activity of osteoblast. The result indicated that the middle dose of the total flovone only could promote the proliferation of the normal osteoblast and the difference was significant（0.01<P<0.05）,but compared to the oxidative damaged cell,the high dose of the total flovone and the flovone both can promote the proliferation and the differentiation of osteoblast,the flavonone didn’t.Thirdly, in the experiments of anti-oxidation with pyrogallic acid in vitro, the active component choosed from the total flovone, the flovone and the flavonone, was the flovone and took the experiment of the mechanism on it.（1）Utilizing H₂O₂ to erect the model of oxidized osteoblast and studing the protection and reparation effect on the osteoblast by detecting the activity of SOD which was affected by the total flovone、the flovonol,the flavonone. The result indicated that the activity of SOD in the group of total flovone and the flovonol was enhanced and the disparity was conspicuous compared to the model group（0.0KP<0.05）.So,the result showed the peroxidation of osteoblast could be prevented by he total flovone and the flovonol.（2）Erecting the pyrogallic acid autoxidation-model to investigate the activity of antioxygen,the flovonol and the corylifolin manifested the prominent activity, and the effect of the flovonol was better to the flavonone.（3）The effect of the flovonol which was the active component of anti-oxidation choosed by the anti-oxidization experiments on the the swelling of oxidative damaged mitochondria and the content of MDA was determined. The result indicated that the flovonol can cut down the swelling and depressed the generation of MDA,the effect was positive correlation to the dose.Finally,HSCCC was adopted in the first time to separate the total flovone and the separation which were flovone and flavonone of Psoraleae were obstanined through initiatory identification,so,the technology of separation to set up the foundation for separating the component from the traditional Chinese medicine. The result indicated the flovone of Psoraleae could promote the cell proliferation and protect the mitochondrion,the mechanism of action probably was related to enhancing the activity of SOD,inhibiting the lipid peroxidation of mitochondrion and lessening the injury to osteoblast and mitochondria caused by the free radical to maintain the normal function of the cell.