| Objective:To screen of the biomarkers of plasma exosome microRNA for early prediction of osteoporosis,it could be used as one of the laboratory methods for early prediction of osteoporosis and has great potential.A new way could be provided for clinicians to diagnose and prevent osteoporosis.Methods:To collect outpatients and inpatients from January 2016 to October 2017in the Endocrinology Department of Xi’an Central Hospital as the research objects.Exclusion of endocrine and metabolic diseases,autoimmune diseases,long-term medication affecting bone metabolism,history of gastrointestinal surgery,liver andkidney dysfunction,malignant blood diseases,hereditary diseases,pregnant,preparation for pregnancy,breast-feeding,poor blood pressure control,severe heart disease,chronic hypoxic disease,condition of stress,poor general condition of the whole body,Age less than 40 and over 76.The diagnostic criteria for osteoporosis are based on the guidelines for the diagnosis and treatment of Primary Osteoporosis published in the ChineseJournal of Osteoporosis and Osteominergic Diseases in 2011 years:Osteoporosis:T≤-2.5;osteopenia:-2.5<T<-1;normal bone mass:T≥-1.21 cases were selected,including osteoporosis（group A,n=6,including severe osteoporosis）,osteopenia（group B,n=10）,normal control group（group C,n=8）.Grouped according to lumbar vertebrae position and femoral neck dual energy X-ray bone mineral density examination,Record age,gender（female also had to record menopausal age）,personal history,medicationhistory of all cases who can be included in the study.Measure their height,weight,systolic and diastolic blood pressure,calculate BMI=weight/height2（kg/m2）,and sign informed consent.Detection of electrolytes（including serum calcium,phosphorus）,liver function（including ALP）,renal function,blood glucose,blood lipid,thyroid function（FT3、FT4、TT3、TT4、TSH）,TPOAb,TgAb,E2,PTH,25（OH）D3,SF etc.in all subjects.Three subjects’plasma were selected from each group.Extract the exosome with Riboexosome Isolation reagent,and separate exosome from plasma,then,extract microRNA,using ND-1000 Nanodrop and Agilent 2200 Tapestation program to inspect samplequality then build the library and sequencing.Using log2ratio and scatter plot to compare the differences in the expression of co-expressed microRNA,EdgeR was used to analyze the significance of microRNA difference among groups,and the P value was calculated.Pearson correlation coefficient r was used to analyze the correlation between group A,B and C,to determine the reliability of sequencing results.Then the miRNAs biomarkers were related to osteoporosis have been confirmed in previous literature.Results:1.Difference and analysis between group A and group C:514 miRNAs were detected in group A and group C,there were 316 miRNAs with high significant difference and 60 miRNAs with general significant difference.37miRNAs were related to osteoporosis have been confirmed in previous literature,of which 17 miRNAs were up-regulated and 20 miRNAs were down-regulated.2.Difference and analysis between group B and group C:735 miRNAs were detected in group A and group C,there were 60 miRNAs with high significant difference and 44 miRNAs with general significant difference.34miRNAs were related to osteoporosis have been confirmed in previous literature,of which 28 miRNAs were up-regulated and 6 miRNAs were down-regulated.3.Relationship between group A and group B:There were 372 miRNAs expressed in group A and group B,39 of which were different from group C and 5 of which with high expression and significant difference.Conclusion:1.Osteoporosis does have differentially expressed miRNAs compared with normal controls.2.The 71 high expression miRNAs were closely related to osteoblasts and osteoclasts,there were 39 miRNAs in group A and group B,which were different from group C.These miRNAs were likely to be a potential biomarker for osteoporosis and osteopenia.However,because of the small sample size and there are individual differences in miRNAs themselves.It is necessary to expand the sample size for further later verification.The fluorescent quantitative PCR method is used to simplify and improve the microRNA expression profile of osteoporosis.3.The expression patterns of the three groups were all positively correlated with each other,which indicated that the detected microRNAs were closely related to the occurrence and development of osteoporosis.4.By detecting these plasma exosome microRNAs,which were screened out from our study,the occurrence of osteoporosis can be predicted in time and the risk of brittle fracture can be reduced.It would be significant for an earlier diagnosis of osteoporosis. |
