Screening Of Osteoporosis Related Inflammatory Factors In Chinese Postmenopausal Women

Osteoporosis(OP)is a systemic chronic metabolic disease characterized by decreased bone density,destruction of micro architecture and increased fragility of bone.It occurs in the elderly people and postmenopausal women.With the accelerated aging of the population,OP and osteoporotic fractures have become a serious public health problem,causing a huge social and economic burden.OP is a complex disease caused by a combination of various factors.At present,the pathogenesis of OP is not completely clear.In this study,high-throughput protein chip technology and cell function experiments were used to screen and validate OP-related inflammatory factors,which aims to find potential biomarkers of OP and to provide scientific evidence for revealing the molecular mechanisms and early prevention of OP.A large number of studies have shown that inflammatory factors play an important role in the development of OP.In view of this,the present study will screen and validate OP-related inflammatory factors and study the molecular mechanisms of inflammatory factors and OP-related cellular functions from the following two parts.In part(?),through using high-throughput protein chip technology,the study will screen the significant differentially expressed inflammatory factors in the plasma samples between the postmenopausal women with extremely high vs.low hip BMD.Then by using ELISA assays,the study will verify and replicate OP-related inflammatory factors in two independent population.In part ?,the study will explore the effect of BLC protein on OP-related bone cells,osteoblasts and osteoclasts,through cell proliferation,cell differentiation,cell apoptosis and cell cycle experiments.Part(?)ObjectiveTo screen the OP-related potential inflammatory biomarkers in Chinese postmenopausal women based on high-throughput protein chip technology and ELISA assays.MethodsBased on a study cohort containing 6,308 Chinese elderly,this study adopted extreme sampling scheme and high-throughput protein chip technology to screen he significant differentially expressed inflammatory factors between the postmenopausal women with extremely high vs.low hip BMD verify and replicate the differentially expressed inflammatory factors in two independent populations.ResultsThe study identified a total of 8 OP-related differentially expressed inflammatory factors.Among them,IGFBP4 and CD97 were upregulated in extremely high BMD,and BLC,M-CSF,IL-11,IL-17,6ckine and OPN were downregulated in extremely high BMD in Chinese postmenopausal women.Taking into account the expression levels of the eight inflammatory factors and the limitations of the present detection methods,this study ultimately selected BLC,IGFBP4 and OPN for further research.Then this study selected validation group(N=39)and replication group(N=41)to verify and replicate whether these three inflammatory factors are related to OP.The results of verification and replication were consistent with the results of screening,that is,BLC and OPN were lowly expressed and IGFBP4 was highly expressed in the extremely high BMD.At the time,the correlation analysis showed that BLC and OPN levels were correlated with BMD,and that IGFBP4 levels was positively correlated with BMD.ConclusionsIn this study,eight OP-related differentially expressed inflammatory factors were screened by using high-throughput protein chip technology.Among them,IGFBP4 and CD97 were upregulated in extremely high BMD,and BLC,M-CSF,IL-11,IL-17,6ckine and OPN were downregulated in extremely high BMD in Chinese postmenopausal women.Then BLC,OPN and IGFBP4 were verified and replicated in two independent populations,and plasma BLC,OPN and IGFBP4 levels were correlated with hip BMD.We proposed BLC,OPN and IGFBP4 to be a potential biomarker involved in the molecular pathogenesis of OP.Part ?ObjectiveTo study the effect of BLC protein on OP-related bone cells and reveal the molecular mechanism of BLC protein on OP based on the contents of the Part(?).MethodsIn this study,MC3T3-E1 cells and RAW264.7 cells were selected as target cells.For MC3T3-E1 cells,RTCA technique for cell proliferation detection,ALP activity staining and real-time PCR for cell differentiation,Annexin V/PI double staining for cell apoptosis detection and cell cycle detection were performed to study the effect of BLC proteins on biological characteristics of osteoblasts.For RAW264.7 cells,RTCA technique for cell proliferation detection,TRAP staining and F-actin staining for the degree of cell differentiation detection,Annexin V/PI double staining for osteoclasts apoptosis detection were performed to study the effect of BLC proteins on biological characteristics of osteoclasts.ResultsFor MC3T3-E1 cells,compared with the blank group,cell proliferation was significant inhibited,cell apoptosis was significantly promoted,ALP activity was inhibited and the expression of COL1A1 and OCN m RNAs were significantly reduced under the effect of BLC proteins,while the cell cycle was not obviously affected.For RAW264.7 cells,compared with the blank group,the cell proliferation was significantly promoted,the number and the volume of osteoclasts induced by RAW264.7 were significantly increased,and the cytoskeletal structure was more complete under the effect of BLC proteins,while cell apoptosis was not obviously influenced.ConclusionsBLC protein may inhibit bone formation by inhibiting the proliferation and differentiation of osteoblasts and promoting apoptosis of osteoblasts.Meanwhile,the protein may promotes bone resorption by promoting the proliferation of pre-osteoclasts and the generation of osteoclasts.The two mechanisms together lead to reduction of bone mass and decrease of BMD,even the occurrence of OP.