Preliminary Study Of The Relationship Among Osteoprotegerin,Receptor Activator Of NF-kB Ligand And Senile Osteoporosis Syndrome Types In Traditional Chinese Medicine

Objective:The aim of this study was explored to the relationships of TCM syndromes, osteoprotegerin(OPG) and Receptor Activator of NF-kB Ligand (RANKL) in senile osteoporosis,in order to find the theory of diagnosis and differentiation of TCM syndromes in senile osteoporosis.Methods:1. First study was designed to determine the effects of the OPG on the mRNA expression of carbonic anhydrase II (CAII) and the receptor activator of NF-kB (RANK) in mouse osteoclast-like cells.Marrow cells Marrow cells were harvested from femora and tibiae of mouse and cultured in6-well chamber slides. After1day of incubation, the marrow cells were exposed to M-CSF (25ng/ml), RANKL(50ng/ml), and different concentrations of OPG (50,75, and100ng/ml, respectively) for3days. The expression of RANK and CAIImRNA was determined with real-time fluorescent quantitative polymerase chain reaction(RT-PCR).2.124old patients came from department of Rehabilitation, Geriatrics and Orthopaedics of our hospital, were selected from Decemeber2010to Decemeber2011. They were diagnosed senile osteoporosis by the bone mineral density (BMD) on anterior posterior position lumbar2-4vertebra body which was measured by dual energy X-ray absorptiometry.According to the principles of the differentiation of syndromes in traditional Chinese medicine,they were divided into four syndrome types,namely kidney-yang deficiency syndrome,Spleen-kidney-yang deficiency syndrome,liver-kidney-yin deficiency syndrome and Qi-deficiency and Blood-stasis syndrome. The OPG and RANKL of serum were detected by ELISA method.The variable inherent relationship of OPQRANKL and BMD in different syndrome types were analyzed with analysis of variance.Results:1. The numbers of multinucleated, TRAP-positive osteoclast-like cells, and resorption pits formed were observed. Compared with the M-CSF+RANKL group, RANK and CAIImRNA expression were statistically decreased in the M-CSF+RANKL+OPG (50ng/ml) groups, M-CSF+RANKL+OPG (75ng/ml) and M-CSF+RANKL+OPG (100ng/ml)(P<0.05, P<0.05,P<0.01respectively).2. Serum OPG was positively correlated to BMD(r=0.194,P<0.05).BMD of kidney-yang deficiency syndrome was higher than other three syndromes (P<0.05,P<0.05,P<0.05respectively).Serum of OPG and RANKL in the Qi-deficiency and Blood-stasis syndrome was lower than kidney-yang deficiency syndrome,Spleen-kidney-yang deficiency syndrome as well as liver-kidney-yin deficiency syndrome, the differences were significiant (P<0.05or P<0.01).Conclusion:1.These data suggest that OPG could regulate the expression of RANK and CAⅡ mRNA in the marrow culture system. OPG,RANKL and RANK play crucial roles and have been identified as essential for differentiation and maintenance of osteoclasts.2. Serum OPG may become predictor of BMD.Serum OPG and RANKL can be used as objective reference index in Qi-stagnation and blood-stasis syndrome distinction of kidney-yang deficiency syndrome, Spleen-kidney-yang deficiency syndrome and liver-kidney-yin deficiency syndrome of senile osteoporosis. The kidney-yang deficiency syndrome may be the early stages of pathogenesis in senile osteoporosis, therefore,treatment should be intervented timely.