The Study Of Gestational Diabetes Mellitus With TCM Syndrome Patterns Based On Metabonomic Data

ObjectiveTo survey the syndrome patterns of gestational diabetic patients and explore various metabolomic biomarkers related to heart yin deficiency,qi yin deficiency or undetermined syndrome pattern,including analysis of metabolic pathways and discern of the specifics of different patterns.To associate the theory and practice of pattern recognition by the example of gestational diabetes mellitus and and seek treatment principles in Traditional Chinese Medicine(TCM).Methods1.A total of 430 cases were studied from patient records between March of 2013 and December of 2016 in the obstetrics department of First Affiliated Hospital of Guangzhou TCM University according to guidelines set forth by Chinese medicine diagnosis group of Chinese Medicine Association,including questionnaires of specifics of Gestational Diabetes Mellitus(GDM)and a summary of common syndrome patterns.2.Sixty GDM cases were divided into 2 groups by gestational age.Group 1(24 weeks to 28 weeks of gestational age)includes 29 cases.Group 2(28weeks to 32 weeks)includes 31 cases.On the basis of syndrome differentiation,there are 20 cases in heart yin deficiency group and 23cases in qi yin deficiency group.In the undetermined group,there are 17cases.The non-diabetic(or OGTT normal)group includes 19 cases in similar gestation age.3.Data from the patients were collected,including maternal age,gestational age,number of gravidity or parity,maternal height,systolic and diastolic pressure,prepregnancy body weight and body mass index(BMI)(BMI=weight[in kilogram]/height~2[in meter])and levels of fasting blood glucose(FBG),one postprandial(1hPG),two hours postprandial(2hPG),glycosylated hemaglobin(HbA1c),fasting insulin(FINS)and two hours postprandial insulin(2hPINS),lipid panel of triglyceride(TG),total cholesterol(CHOL),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),hemaglobin and thyroid function of thyroid stimulating hormone(TSH),free thyroxine(fT4),antithyroidperoxidase antibody(TPO Ab).4.Serum samples from fasting patients were collected and studied for M etabolomic profiling by ultra-performance liquid chromatography/quadrupol e-time-of-flight mass-spectrometry(UPLC/Q-TOF-MS)method using Masslynx V4.1 workstation to collect the data,importing Progenesis QI 2.0 and ana lysing data by EZinfo software.To detect various metabolomic biomarkers b y different pattern recognition methods,such as PCA?PLS-DA?OPLS-DA,wer e used for analysis and comparison from HMDB,KEGG and other databases an d literature to determine the different biomarkers,and analysis the meta bolic pathways.Results1.Frequencies of the common manifestation are listed:insomnia(85.0%),forgetfulness(83.8%),frequent urination(81.2%),dizziness(79.3%),lumbar discomfort(78.8%),spontaneous perspiration(71.3%),edema(58.5%),pale tongue with tooth marks(55.4%),fatigue(51.6%),thin and rapid pulse(50.9%),dry stool(45.4%),thin white tongue coating(1.6%).Common TCM syndromes patterns include yin deficiency,qi yin deficiency,spleen kidney Yang deficiency,spleen deficiency and dampness,kidney Yin deficiency.In cases of the scores below 14 are assigned to undetermined pattern.2.Data comparison:Difference between 2 gestational periods is not statistically significant.Similarly,differences are not statistically significant among 4 groups of heart yin deficiency,qi yin yin deficiency,undetermined group and the control with regard to maternal age,gestational age,number of gravidity or parity,maternal height or body mass index.3.Comparisons of other the clinical data:Compared with the(normal)control group,the FBG,1hPG,2hPG,Hgb levels are elevated in the GDM patients and AST levels decreased;the differences are statistically significant(P<0.05).Group 2(28-32 weeks)showed increased HbA1c and TG levels;the differences are statistically significant(P<0.05).Yet no significant difference for TSH,fT4,TPO Ab,CHOL,TG,HDL-C,LDL-C between Group 2 and Group 1(24-28 weeks)(P>0.05).Heart yin deficiency,qi yin deficiency and undetermined group showed no statistical differences in FPG?1hPG?2hPG?HbA1c?FINS?2hPINS?HOMA-IR?ISI?HOMA-?(P>0.05).Differences of Hb,TSH,fT4,TPO Ab,CHOL,TG,HDL-C,LDL-C,AST,ALT are not statistically significant(P>0.05).4.Pattern recognition method by Partial Least Squares-Di scriminant Analysis(PLS-DA)and Optimized Potentials for Liquid Simulations discriminant analysis(OPLS-DA)showed discriminated groups of heart yin deficiency,Qi Yin deficiency,undetermined syndrome pattern and the normal(control).The cumulated values of each group are mainly distributed in the elliptical scatter plot(95%confidence interval,CI)without any cross and overlap.5.After screening,a total of 25 compounds were found to be associated with different gestational weeks of GDM and Chinese medicine syndrome patterns.Compared with group 1(24-28 gestational weeks),group 2(28-32gestational weeks)displayed a rising trend for 11 metabolites and the differences of 8 metabolites were statistically significant(P<0.05),indicating that blood metabolite profile changes occurred during different gestational periods.6.Compared with the control group,heart yin deficiency group showed a rising trend for 14 metabolites and the differences of 12 metabolites showed rising trend statistically significant(P<0.05).Qi yin deficiency group showed a rising trend for 12 metabolites,yet 2 metabolites were reduced statistically significant(P<0.05),levels of only 6 metabolites were statistically different(P<0.05).Undetermined syndrome pattern group showed13 metabolites increased and one metabolite decreased.The data indicated all3 GDM pattern patients showed variations of changes when compared with the control group,particularly the apparent deviation in the heart yin deficiency group.7.Comparing profiles among all 3 GDM patterns,thirteen metabolites showed rising trend and the 12 of these were significantly different(P<0.05),The results indicated all 3 GDM pattern patients again showed variations of changes.8.In two different types of grouping groups,molecular masses of 782.5688,758.5691,730.5892,544.3442 metabolites are different.After identification,the differential metabolites were phosphatidylcholine(PC)(36:4),PC(34:2),phosphatidylethanolamine(20:1)and lysophos-phatidylcholine(22:4).Conclusion1.Most diabetic patients are considered deficient in nature.Deficiency is combined with qi deficiency and dampness.Heart,spleen and kidney are the Zangfu involved.Main patterns are heart yin deficiency and qi yin deficiency.Also our previous studies reported dampness and spleen deficiency frequently occurred among older women and severe insulin-resistance.In clinical practice,it is a good idea to strengthen the spleen and drain the dampness to mitigate insulin-resistance.2.Three different patterns of GDM patients showed no statistical differences in most conditions,including the liver,thyroid functions etc.The same were the the blood glucose and lipid levels,suggesting other biomarkers may be explored to differentiate the different syndrome patterns.3.Variations of metabolomic profiles among GDM patients indicated the metabolite variations in the study,particularly the differences amon g d i f f e r e n t g e s t a t i o n a l a g e a n d t h e s y n d r o m e p a t t e r n s.4.Levels of phosphatidylcholine(PC 36:4),(PC 34:2),lysophos-phatid ylcholine(LPC 22:4),phosphatidylethanolamine PE(20:1)may be the metabo lites associated with different GDM TCM syndrome patterns.They may be ap plied to screening,intervention and management of GDM in the first trime ster of pregnancy.