Left Youguiwan, On Cultured Rat Osteoblasts

Object:To observe the influence of ZuoGui pill and Yougui pill,on proliferation of osteoblast,generation of Estrogen and mRNA expression of osteoblast aromatase.Method:Direct medication:The solution of Zuogui pill,yougui pill,and their herb-sorted groups at the concentration of 100\10\1\0.1\0.01ug/ml is taken respectively to culture osteoblast.After 48 hours culturing,the inhibition ratio or the strength and weakness of OB proliferation was counted according to the volume of light-absorption.The solution of Zuogui,yougui,and the herb-selected groups at the concentration of of 50ug/ml with additional testosterone of 1mm was applied to culture osteoblast.After 48hs or 72hs culturing,the supernatant of culture solution was taken to examine the volume of E2.The solution of Zuogui,yougui,and the herb-sorted group at the concentration of of 50ug/ml was applied to culture osteoblast.After 8h and 24h culturing,the general RNA of the cultured cells was extracted to determine the aromatase expression by RT-PCR.Medicine-serum medication:90 female mice were divided into 6 groups,such as control group,Zuogui pill group,Yougui pill group,No.1 herb-sorted group,No.2 herb-sorted group,and No.3 herb-sorted group.To feed them physiologic saline, decoction of Zuogui pill,decoction of You gui pill and the decoction of the three herb-sorted groups respectively,the femoral artery blood was got from each three mouse at 0.5h,2h,4h,8h,24h,thus to acquire normal serum of physiologic saline,Zuogui pill,Yougui pill,the three herb-sorted groups.The normal mouse serum at the concentration of 1/20,1/200,1/2000,and the serum of Zuogui,Yougui and No.1.No.2.No.3 herb-sorted group at concentration of 1/20 acquired after different time medication were applied to culture osteoblast.After 48 hours culturing, the inhibition ratio or the strength and weakness of OB proliferation was counted according to the volume of light-absorption.The serum was acquired after different time medication of Zuogui pill,yougui pill,and the herb-sorted group at the concentration of 1/20 with additional testosterone at the final concentration of 1mm to culture OB.after culturing 48 hs and 72 hs,the supernatant of culture solution was taken to determine the volume of E2.The serum was acquired after the 8hs and 24hs medication of Zuogui pill,yougui pill,and the herb-sorted groups at the concentration of 1/20.After 24 hours culturing,the general RNA of the cultured cells was extracted to determine the aromatase expression by RT-PCR.Results:1.Direct Medication Zuogui pill:At the concentration of 100ug/ml,it can promote the proliferation with little effect on synthesis of E2,and it can promote mRNA expression of Aromatase at the concentration of 50μg/mL.Yougui pill:It shows weak effect and can inhibit the synthesis of E2,and it can promote mRNA expression of Aromatase at the concentration of 50μg/mL.2.Medicine-serum Medication Zuogui pill:The serum acquired at different time has no obvious action on proliferation of OB.The serum of 4hs and 24hs can promote the synthesis of E and the serum of 24hs can inhibit the mRNA expression of OB aromatase.Yougui pill:The serum acquired at different time has no obvious action on proliferation of OB.The serum of 4hs and 24hs can promote the synthesis of E2 and the serum of 24hs can inhibit the mRNA expression of OB aromatase.Conclusion:To promote estrogen synthesis of osteoblast is the internal functional mechanism of Zuogui pill and Yougui pill to strengthen bone through tonifying kidney.