Correlation Between Osteoporosis And Lymphocyte Subsets In Peripheral Blood And Bone Marrow Of Postmenopausal Patients With Hip Replacement

Background:Postmenopausal osteoporosis(PMOP)is a systemic disease characterized by osteopenia and changes in bone microarchitecture,primarily due to marked decreases in postmenopausal estrogen levels.In recent years,the effect of immune regulation on PMOP has gradually become a ho t research topic.Studies have shown that immune cells can affect bone homeostasis through intercellular contact or paracellular secretion mechanism,and then trigger the decline of b one mass in OVX mice.Objective:In this study,we aimed to understand the correlation between lymphocyte subsets and bone mass in postmenopausal population by analyzing the clinical data such as bone marrow and peripheral blood lymphocyte subsets and bone mineral density,so as to provide clinical data for further research.Methods:1.Overall plan:(1)postmenopausal patients undergoing hip joint replacement in the joint surgery department of the second affiliated hospital of suzhou university were collected according to the nano-exclusion standard as the research subjects,peripheral blood samples were taken from all the research subjects after admission,bone marrow samples at the proximal femur during operation and bone mineral density(dual-energy x-ray absorptiometry)after operation.(2)Flow cytometry analysis:①The proportion of lymphocyte subsets in peripheral blood and bone marrow;② The absolute counts of lymphocyte subsets in peripheral blood and bone marrow;③The proportion of bone marrow T lymphocyte subsets.SPSS and Graphpad software were used to an alyze the correlation between bone mineral density and lymphocyte subsets.2.Specific grouping:(1)first stage:proportion of lymphocyte subsets.First of all.30 subjects were included in the.study to investigate the proportion of lymphocyte subsets.They were divided into the osteoporosis group,the osteopenia group and the normal bone mass group according to the bone mineral density results,and relevant flow cytometry analysis and statistics were conducted.(2)The second stage:the absolute count of lymphocyte subsets.Based on the results of the first stage,which showed that the proportion of lymphocyte subsets was related to the bone m ass of postmenopausal population,27 subjects were included again,and they were analyzed by the proportion of lymphocyte subsets and absolute count.According to the bone mineral density results,they were divided into the osteoporosis group,the osteopenia group and the normal bone mass group,and relevant flow cytometry analysis and statistics were performed.(3)The third stage:the proportion of bone marrow T lymphocyte subsets.Based on the results of the Phase Ⅰ and Ⅱ studies,which showed that bone marrow T lymphocytes had the highest correlation with bone mass in postmenopausal population,44 subjects were included again for further study,and the proportions of lymphocyte subsets and absolute counts in peripheral blood and bone marrow as well as the proportions of bone marrow T lymphocytes subtypes were analyzed.The subjects were divided into the osteoporosis group,the osteopenia group and the normal bone mass group according to the bone mineral density results,and the relevant flow cytometry analysis and statistics were conducted.(4)The fourth stage:the analysis of similar data integration.A total of 101 subjects were included in the first three stages and 71 subjects were included in the second and third stages for absolute count of lymphocyte subsets.Data integration analysis was performed.(see P3-4 for detailed process and grouping)Results:1.first Stage:The proportion of lymphocyte subsets.Correlation analysis showed that the proportion of bone marrow CD3+T lymphocytes(CD3)(R=-0.527,P=0.003).and the proportion of bone marrow CD4+T lymphocytes(CD4)(R=-0.497.P=0.002)had a negative correlation with bone mass in postmenopausal population.After grouping by bone mineral density,there were significant inter-group differences in bone marrow CD3 proportion(P=0.009)and bone marrow CD4 proportion(P=0.005).Linear regression analysis showed that bone marrow CD3 proportion(P=0.003)and bone marrow CD4 proportion(P=0.002)had linear relationship with bone mass of postmenopausal population.2.The second stage:absolute count of lymphocyte subsets.Correlation analysis showed that absolute CD3 count in peripheral blood(R=0.572,P=0.002)and absolute CD3 count in bone marrow(R=-0.536.P=0.004)were negatively correlated with bone mass in postmenopausal population.After grouping by bone mineral density,there were significant inter-group differences in the absolute CD3 count in peripheral blood(P=0.006)and bone marrow(P=0.024).Linear regression analysis showed that absolute CD3 count in peripheral blood(P=0.0475)and absolute CD3 count in bone marrow(P=0.0328)had linear relationship with bone mass in postmenopausal population.3.The third stage:Percentage of bone marrow T-lymphocyte subsets.Correlation analysis showed that the proportion of bone marrow regulatory T lymphocytes(Treg)(R=0.401.P=0.007)was positively correlated with bone mass in postmenopausal population.The proportion of bone marrow Treg(P=0.005)showed significant inter-group difference after grouping by bone mineral density.Linear regression analysis showed that the proportion of bone marrow Treg(P=0.003)had a linear relationship with the bone mass of postmenopausal population.The results of ROC curve and Jordan index suggested that the area under the Treg curve of bone marrow(0.815)was greater than 0.7,with statistical significance.4.The fourth stage:integration analysis of similar data.According to the ROC curve and Jordan index results,it was suggested that the area under the curve of bone marrow CD3 proportion(0.883),bone marrow CD4 proportion(0.772),absolute peripheral blood CD3 count(0.830)and absolute bone marrow CD3 count(0.847)were greater than 0.7,which was statistically significant.The other results were basically consistent with those in the previous study.Conclusions:The above conclusions preliminarily indicate the correlation between the reduction of bone mineral density in postmenopausal population and abnormal immune indexes,so further exploration of the related mechanisms and prediction scheme of immune regulation and PMOP in the future will have certain reference value for clinical work.