Effect Of The Total Flavonoids Of Psoralea And Herba Epimedii On Proliferation And Antioxygen Of Osteogenesis Cells And Kidney Cells

The Psoraleae belongs to leguminous plant and is the fruit of the plant.It is warm-natured and acrid flavor,besides invigorating the kidney to promote masculine elements.It was adopted to cure the disease such askidney deficiency,cold diarrhea,invirility,preventing bone resorpted,increasing bone density,asthenia cold,anddyspnea with cough in the orthodox clinic of traditional Chinese physician.The modern studies indicate that psoraleae can distend blood vessel,raise the myocardial contractile force,antitumor,antisepticise and so on.As yet,the professional have got about forty compounds of coumarins,flavonoid and terpene from Psoraleae.The compounds of flavonoid and coumarins are important ingredients which can display the pharmaco-acticity.The Epimedii belongs to Epimedium herb.Shizhen Li said that it has tonifying didney genuine energy and srenthening the bones and muscles and strengthening waist and hness and strengthening cardiac efficiency in Compendium of Materia Medica.The modem studies indicate that Epimedii can improve blood supply to heart and brain,improve haematogenesis and immune function and bones metabolism,anti-aging,anti-tumor and so on.The study of The Epimedii flavone is not penetrating.Some experts study compound recipe of Chinese crude drug deeply recently.They not focus on Substructure of Chemical materials and Compatibility mechanism but Pharmacology and Clinical Curative effect.We will research Substructure of Chemical materials by modern technology for Clinical.Firstly,the High Speed Countercurrent Chromatography（HSCCC） is adopted to purify the total flovone from Psoraleae and Epimedii and Compound recipe.In this experiment,we establish the technology to separate and purify initially.Through the experiment,the solvent system of Psoraleae was applied which was composed with N-hexane-acetic ether-methanol-water in proportion of 1:1:1:1;the solvent system of Epimedii was applied which was composed with chloroform-methanol -water in proportion of 4:3.3:2;the solvent system of Compound recipe was applied which was composed with N-hexane-acetic ether-methanol-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 extraction was 25℃and the rotation speed was 800r/min,then,we got some peaks by detected with 254nm ultraviolet,each peak represents one component.Then,the chemical properties of 6 components were identified initially from Psoraleae and 4 components were identified initially from Epimedii and 5 components were identified initially from Compound recipe. We affirm that the Psoraleae component of peakⅣandⅤare flavonol and flavonone;the Epimedii component of peakⅡandⅢare icariside and icariin,the Compound recipe component of peakⅢis flavone.The second,the expriment on normal osteoblast was carried out to detect the effect of the total flovone of Psoraleae,Epimedii and Compound recipe on the cell proliferation.The result indicated that the middle dose(1.0×10^-7g/L) of the total flovone of Psoraleae,Epimedii,Compound recipe could promote the proliferation of the normal osteoblast and the difference was significant （0.01＜P＜0.05）;but compared to the oxidative damaged osteoblast,the high dose(1.0×10^-6g/L) and the middle dose(1.0×10^-7g/L) of the total flovone Psoraleae and Epimedii,the high dose (1.0×10^-6g/L) of Compound recipe all can promote the proliferation and the difference was significant with the sample（0.01＜P＜0.05）.The third,the expriment on normalkidney cells were carried out to detect the effect of the total flovone of Psoraleae,Epimedii and Compound recipe on the cell proliferation.The result indicated that the middle dose(1.0×10^-7g/L) of the total flovone of Psoraleae,Epimedii,Compound recipe and the high dose(1.0×10^-6g/L) of Epimedii could promote the proliferation of the normalkidney cells and the difference was significant（0.01＜P＜0.05）;but compared to the oxidative damagedkidney cells,the high dose(1.0×10^-6g/L) of the total flovone Psoraleae and Compound recipe both can promote the proliferation and the difference was significant with the sample （0.01＜P＜0.05）.The fourth,through the experiments on antioxygen in vitro,the capability of anti-oxidation of the total flovone of Psoraleae,Epimedii and Compound recipe were detected.（1） Utilizing H₂O₂ to erect the model of cellular oxidation and studied the total flovone of Psoraleae,Epimedii,Compound recipe on the aetivity of superoxide dismitase（SOD） and the content of malonaldehyde（MDA）.The activity of SOD enhanced and the content of MDA decreased in the group of the total flovone of Psoraleae and Compound recipe.The difference was significant with the sample（0.01＜P＜0.05）.The total flovone of Psoraleae and Compound recipe can improve the ability cells to resist oxidant injury.（2） Erecting the pyrogallic acid autoxidation-model to investigate the activity of antioxygen,the total flovone of Psoraleae and manifested the prominent activity,and the effect of the Compound recipe was better to the Psoraleae.The last was Epimedii.Finally,the HSCCC was adopted to separate the total flovone of Psoraleae,Epimedii, Compound recipe and got the components of flavonol and flavonone from Psoraleae and icariside and icariin from Epimedii and flavone from Compound recipe by initially identified,so,the technology of purifying to set up the foundation for purifying the component from the traditional Chinese medicine.The result displayed the total flovone of Psoraleae,Epimedii,Compound recipe could promote the cell proliferation,the mechanism of action probably is lessening the damaged to cells caused by the free radical and the activity of SOD enhanced and MDA decreased.