Effects Of ZhuangGuZhen Tong Capsule Contained Plasma On Activity And Apoptosis Of Osteoclast In Osteoblasy And Osteoclast Co-culture System

Objective:To study effects of the ZhuangGuZhenTong Capsule (ZGZTC)-containing plasma on activity and apoptosis of osteoclast,and to study the mechanism of action of ZGZTC in the prevention and treatment of bone metastases of breast cancer from the way of plasma pharmacology.Methods:1.Establishing the plane of osteoblast and osteoclast co-culture system with agarose gel and 0.45μm aperture microporous membrane.2.Female SD rats aged 10 month were divided into ZGZTC groups and control group (normal physiological saline) to prepare ZGZTC-containing plasma and control plasma.3.Determine the effect of 2.5% ZGZTC-containing plasma on absorption of lacuna formed by osteoclast (OC) in osteoblast and osteoclast co-culture system by toluidine blue staining.4.Determine the effect of 2.5% ZGZTC-containing plasma on activity of TRAP secreted by OC in osteoblast and osteoclast co-culture system by the diazol method.5.Determine the effects of 2.5% ZGZTC-containing plasma on OC survival and apoptosis in osteoblast and osteoclast co-culture system by reverse and fluorescence microscopes.Result:1.Protein dispersing experiment:after detection by Coomassie Brilliant blue staining,we demonstrated that the protein in osteoblast(OB) culture room dispersed to OC culture room through membrane within 8 hours, which suggested that the establishment of plane osteoblast and osteoclast co-culture system was successful.2.The number of absorption of lacuna of 2.5%ZGZTC-containing plasma group was obviously less than that of the control group in the osteoblast and osteoclast co-culture system(p<0.01).3.The numbers of TRAP secreted by OC in the osteoblast and osteoclast co-culture system of 2.5% ZGZTC-containing plasma group at 48h,72h and 96h showed an obvious time-efficiency relationship,which were remarkably lower than that of the control group(p<0.01)4.The OC surving numbers in the osteoblast and osteoclast co-culture system of 2.5% ZGZTC-containing plasma group in 48h,72h and 96h showed an obvious time-efficiency relationship,which were also remarkably lower than that of the control group(p<0.01).The OC apoptosis numbers in the osteoblast and osteoclast co-culture system of 2.5% ZGZTC-containing plasma group in 48h,72h and 96h had an obvious time-efficiency relationship,which were remarkably higher than that of the control group(p<0.01)Conclusion:The ZGZTC-containing plasma:significantly suppressed the activity of OC and improved the withering of OC.