Clinical Metabolomics Of Primary Osteoporosis Based On Yin/Yang Deficiency Syndrome

Objective:Using metabolomics technology,the biological marker characteristics of kidney Yang deficiency syndrome and liver and kidney Yin deficiency syndrome in primary osteoporosis were explored,so as to provide research evidence for the microscopic syndrome differentiation and syndrome efficacy evaluation of kidney Yang deficiency syndrome and liver and kidney Yin deficiency syndrome in primary osteoporosis.At the same time,the correlation between metabolic markers and clinical indicators was studied to guide clinical practice.Method:1.According to the diagnosis of primary osteoporosis screening of patients,according to standard of kidney Yang deficiency syndrome and liver and kidney Yin deficiency syndrome,from group collection in patients with primary osteoporosis screening of 30 cases of kidney Yang deficiency syndrome syndrome is the most typical female patients,30 cases of liver and kidney Yin deficiency syndrome and syndrome is the most typical female patients,control of 60 patients with baseline for 30 cases of healthy subjects in postmenopausal women.2.General information items(including age,weight,height,blood pressure,pulse,etc.)were collected.Double-energy X-ray lumbar bone density(BMD)data were recorded.Fasting blood was taken from the subjects and biochemical indexes of bone metabolism were determined.3.Subjects1 fasting blood was extracted and tested by LC-MS Metabolomic detection was performed,and UNIFI 1.8.1 was used.The software was used to collect the original data.The original data were subject to baseline filtering,peak recognition,integration,retention time correction,peak alignment and normalization by Progenesis QI(v2.3),The Metabolome processing software.The Human Metabolome Database(HMDB),Lipidmaps(v2.3)and METLEN Database were used for qualitative analysis,and then The data was imported into The metabolomics data analysis software SIMCA-P for analysis.Using the method of univariate analysis and multivariate analysis to conduct statistical analysis,and then to screen the potential metabolic markers of kidney Yang deficiency syndrome and liver and kidney Yin deficiency syndrome in primary osteoporosis.4.Combined with the data of metabolic markers,statistical correlation analysis was conducted on metabolic markers,basic information of subjects and various clinical indicators.Result:1.All the data were in accordance with normal distribution by Kolmogorov-Smirnov test(P>0.05).2.General data results:according to F test,there was significant difference in age among three groups(P<0.05).By t-test,there was no significant difference in age between kidney yang deficiency group and liver kidney yin deficiency group(P>0.05).There was no significant difference in BMI among three groups(P>0.05).3.Results of clinical indicators:according to the F test,there were differences in bone mineral density among the three groups(P<0.05).There was no significant difference between the two groups(P>0.05).There was no significant difference in 250HD,tp1np and PTH among three groups(P>0.05).By F test,there was significant difference in OC and β-CTX between three groups(P<0.05).By t-test,there was no significant difference between the two groups(P>0.05).4.Metabonomics results:the serum full spectrum metabonomics test showed that the 9 metabolic markers related to primary osteoporosis were 4-trimethylammonium-butyrate-2 enol,inosine,DG(15:0/18:3(9z,12z,15z)/0:0),DG(15:0/18:4(6Z,9z,12z,15z)/0:0),PS(18:0/18:1(9z)),phenylalanine isoleucine,1,2-dehydrated white sunflower,TG(8:0/10:0/a-13:0)[RAC],10 methyl palmitic acid.Among these molecules,4-trimethylammonium-butyrate-2 enol,DG(15:0/18:3(9z,12z,15z)/0:0),DG(15:0/18:4(6Z,9z,12z,15z)/0:0),phenylalanine-isoleucine,TG(8:0/10:0/a-13:0)[RAC]showed a downward trend in primary osteoporosis,and the difference multiples were all below 0.5 times,which was statistically significant compared with the healthy control group(P<0.05).Inosine,PS(18:0/18:1(9z)),1,2-dehydrated white sunflower,10-methyl-palmitic acid showed an upward trend,and the difference multiple was more than 2 times,which was statistically significant compared with the healthy control group(P<0.05).The results of serum full spectrum metabonomics showed that L-phenylalanine and s-lactose glutathione were the two metabolic markers related to kidney yang deficiency and liver kidney yin deficiency in primary osteoporosis.L-phenylalanine was up-regulated in patients with primary osteoporosis(kidney yang deficiency and liver kidney yin deficiency).Compared with the healthy control group,the difference multiple was more than 2 times,the difference was statistically significant(P<0.05).Compared with the group of deficiency of liver and kidney yin,L-phenylalanine was up-regulated in the patients with defnciency of kidney yang,the difference multiple was more than 2 times,the difference was statistically significant(P<0.05).S-lactose glutathione in primary osteoporosis patients with kidney yang deficiency showed an upward trend;compared with the healthy control group,the difference multiple was more than 2 times,the difference was statistically significant(P<0.05).Compared with the healthy control group,the difference was less than 0.5 times,the difference was statistically significant(P<0.05).Compared with the liver kidney yin deficiency group,s-lactose glutathione was up-regulated in the kidney yang deficiency group,the difference multiple was more than 2 times,the difference was statistically significant(P<0.05).5.Correlation results:according to Pearson analysis,BMD was negatively correlated with age,tp1np,OC and β-CTX.(r=-0.49,-0.21,-0.40,-0.36);no correlation with BMI,250HD,PTH(r=0.19,-0.10,-0.01).According to Pearson’s analysis,bone mineral density and 4-trimethylammonium-butyrate-2 enol,DG(15:0/18:3(9z,12z,15z)1 0:0),DG(15:0/18:4(6Z,9z,12z,15z)/0:0),phenylalanine isoleucine,TG(8:0/10:0/a-13:0)[RAC]were positively correlated(r=0.35,0.34,0.29,0.5,0.26);they were negatively correlated with inosine,PS(18:0/18:1(9z)),1,2-dehydrated white sunflower,10-methyl-palmitic acid,L-phenylalanine,s-lactose glutathione(r=-0.25,-0.37,-0.32,-0.39,-0.44,-0.26).According to Pearson’s analysis,subjects,age and 4-trimethylammonium-butyrate-2 enol,DG(15:0/18:3(9z,12z,15z)/0:0),DG(15:0/18:4(6Z,9z,12z,15z)/0:0),phenylalanine isoleucine,TG(8:0/10:0/a-13:0)[RAC]were negatively correlated(r=-0.3,-0.39,-0.31,-0.42,-0.32);they were positively correlated with inosine,PS(18:0/18:1(9z)),1,2-dehydrated white sunflower,10-methyl-palmitic acid,L-phenylalanine,s-lactose glutathione(r=0.28,0.34,0.25,0.36,0.39,0.25).According to Pearson analysis,tp1np was positively correlated with inosine(r=0.26).According to Pearson analysis,PTH was positively correlated with DG(15:0/18:3(9z,12z,15z)/0:0),TG(8:0 1 10:0/a-13:0)[RAC](r=0.21,0.26).According to Pearson analysis,there was a positive correlation between OC and 1,2-dehydrated white sunflower(r=0.22).According to Pearson analysis,there was a positive correlation between β-CTX and L-phenylalanine,s-lactose glutathione(r=0.39,0.25).Conclusion:1.With the increase of age,bone mass gradually decreased,and the incidence of primary osteoporosis increased.2.Among the biochemical indexes of bone metabolism,tp1np,OC and β-CTX are three markers of bone turnover,which have good effect on the evaluation of osteoporosis.At the same time,β-CTX may be related to the classification of kidney yang deficiency and liver kidney yin deficiency in primary osteoporosis.3.The potential serum metabolic markers of primary osteoporosis were 4-trimethylammonium-butyrate-2 enol,inosine,DG(15:0/18:3(9z,12z,15z)/0:0),DG(15:0/18:4(6Z,9z,12z,15z)/0:0),PS(18:0/18:1(9z)),phenylalanine isoleucine,1,2-dehydrated white sunflower,TG(8:0/10:0/a-13:0)[RAC],10 methyl palmitic acid.4.It was found that amino acids and their analogues,including L-phenylalanine and s-lactose glutathione,were the potential markers of serum metabolism in the kidney yang deficiency group and liver kidney yin deficiency group of primary osteoporosis.5.Cystine may be negatively correlated with bone metabolism.6.We speculated that tp1np and inosine,PTH and DG(15:0/18:3(9z,12z,15z)/0:0),TG(8:0/10:0/a-13:0)[RAC],OC and 1,2-dehydrated white sunflower hormone,β-CTX and L-phenylalanine,s-lactose glutathione may have metabolic pathway correlation.