Study On The Effect Of Serum Of Kidney Yang Deficiency Rats On The Expression Of Osteoporosis Related Proteins β-catenin,OPG And RANKL Based On OB-OC Co-culture System

Objective:Objective to probe into the influence of serum of kidney yang deficiency rats on the deliver of osteoporosis related proteinsβ-catenin,OPG and RANKL by 0B-OC co-culture system,and further make a thorough inquiry the possible mechanism of osteoporosis induced by serum of kidney yang deficiency rats.Methods:16 male SD rats were stochastic assigned as blank group and model group with 8 rats in each group.The blank group was rendered normal saline by gavage,and the model group was given the same amount of 10ml·kg^-1 adenine by gavage to establish the kidney yang deficiency model.When building model was finished,test the content of serum c AMP and c GMP in rats,If the model was established successfully,Then gather the serum.In vitro isolating culture the OB and OC,alizarin red,ALP evaluation OB,observe its shape through the Giemsa staining,trap staining evaluation OC,OB-OC co-culture system was established by Transwell chamber,DKK-1 drug group was used as positive group,icariin group(100μmol·L^-1),blank group,icariin(100μmol·L^-1)+serum group（10%kidney yang deficiency serum）,serum group（10%kidney yang deficiency serum）and DKK-1 drug group were set up,After intervention for 2 days,number of OC was counted in each group of cells,ALP and trap contents in supernatant were detected by microplate enzyme labeling method,collect the protein of each group,and WB testing expressions ofβ-Catenin,OPG and RANKL protein.Results:1.Identification results of kidney yang deficiency model:After modeling,the rats in the model group showed obvious manifestations of kidney yang deficiency,such as cold limbs and sparse hair;At the same time,compared with the blank group,the content of camp in the model group decreased（P<0.01）,while the content of c GMP increased（P<0.01）.Combined with the symptoms of rats and the results of plasma indicators,we can identify the success of the model.2.Cell experiment results:compared with blank group,ALP activity,OPG andβ-catenin protein expression in serum group were reduced（P<0.05）,while OC number,TRAP activity and RANKL protein expression were go up（P<0.05）.Compared with serum group,ALP activity in icariin group was go up,TRAP activity and protein RANKL were reduced（P<0.01）,ALP activity and OPG protein expression in icariin+serum group were reduced The activity of trap and the expression of RANKL were reduced（P<0.05）.Conclusions:The serum of kidney yang deficiency model can induce OP,and repress ALP activity,control the expression ofβ-catenin and OPG protein,enhancing the activity of trap and raising the expression of RANKL protein maybe its machine-processed.