A Study For Saliva Of Type 2 Diabetic Patients With Chronic Periodontitis Based On Untargeted Metabolomics

Objectives: To investigate the biomarker metabolites and metabolic pathways in non-stimulated whole salivas of patients with type 2 diabetes mellitus with chronic periodontitis,and to explore the possible role of these metabolites in the bidirectional relationship between periodontitis and type 2 diabetes mellitus.Methods: We selected patients with periodontitis in the Hospital of Stomatology Kunming Medical University from July 2021 to December 2022.According to the inclusion and exclusion criteria,6 patients with type 2 diabetes mellitus with chronic periodontitis and 53 patients with chronic periodontitis were screened,and 24 periodontal healthy volunteers were recruited as the research objects.According to the sampling standard collect whole unstimulated saliva(WUS),Then the Ultra Performance Liquid Chromatography Tandem Mass Spectrometry(UPLC-MS/MS)detection platform was used for metabolic analysis.Based on the quantitative information of metabolites obtained by database matching,The differential metabolites and metabolic pathways were screened by statistical analysis.Results: 1.The type 2 diabetes with chronic periodontitis group(T2D group),chronic periodontitis group(CPD group)and healthy control group(NC group)showed good clustering in the OPLS-DA model,which could be effectively distinguished after pair comparison,and there were differences in the composition and content of metabolites.2.We screened for 27 differential metabolites associated with type 2 diabetes with chronic periodontitis,including: calcitriol,calcifediol,L-palmitoylcarnitine,Lacetylcarnitine,phenylacetamide,salicylaldehyde,corticosterone,12-Oxo ETE,15-Oxo ETE,narbonolide,nattridole,eicosapentaenoic acid,linoleic acid,riboflavin,hexadecanedioic acid,aldosterone,(25s)-7-dafachronic acid,indoleacetic acid,all trans-retinal,estrone,17beta-Estradiol 3-(beta-D-glucuronide),sedoheptulose 7-phosphate,uric acid,5-L-glutamyl-L-alanine,guanine,aspartame,epicatechin.After enrichment of differential metabolites,38 metabolic pathways closely related to type2 diabetes with chronic periodontitis were screened out,including: steroid biosynthesis,tuberculosis,steroid hormone biosynthesis,parathyroid hormone synthesis,progesterone,androgen and estrogen receptor agonists/antagonists,fatty acid degradation,insulin resistance,fatty acid metabolism,styrene degradation,naphthalene degradation,prion diseases,prolactin signaling pathway,lipopolysaccharide biosynthesis,carbon fixation in photosynthetic organisms,arachidonic acid metabolism,endocrine and other factors regulate calcium reabsorption,biosynthesis of12-,14-and 16-membered macrolides,opioid receptor agonists/antagonists,bile secretion,biosynthesis of unsaturated fatty acids,glutathione metabolism,riboflavin metabolism,ovarian steroidogenesis,pentose phosphate pathway,cutin,suberine and wax biosynthesis,aldosterone-regulates sodium reabsorption,longevity regulating pathway-worm,plant hormone signal transduction,retinol metabolism,biosynthesis of nucleotide sugars,carbon metabolism,microbial metabolism in diverse environments,taste transduction,biosynthesis of amino acids,flavonoid biosynthesis,phenylpropanoid biosynthesis.These enriched pathways were involved in lipid metabolism,glucose metabolism,vitamin metabolism,amino acid metabolism,nucleotide metabolism,oxidative stress,endocrine system,etc.Conclusions:1.In the context of existing studies,19 potentially landmark metabolites of type 2 diabetes mellitus with chronic periodontitis are highlighted,including: Calcitriol,L-palmitoylcarnitine,estrone,L-acetylcarnitine,phenylethylamide,12-Oxo ETE,15-Oxo ETE,eicosapentaenoic acid,linoleic acid,riboflavin,epicatechin,cedroheptulose 7-phosphate,5-L-glutamyl-L-alanine,uric acid,corticosterone,aldosterone,prion protein and indoleacetic acid.2.We discussed 25 metabolic pathways related to the screening of differential metabolites,including: lipid metabolism related to steroid biosynthesis,steroid hormone biosynthesis,parathyroid hormone synthesis,secretion and action,progesterone,androgen and estrogen receptor agonist/antagonist,aldosterone-regulated sodium reabsorption,fatty acid degradation,ovarian steroidogenesis,fatty acid metabolism,prolactin signaling pathway,biosynthesis of unsaturated fatty acids,and arachidonic acid metabolism.The pathways related to glucose metabolism include lipopolysaccharide biosynthesis,biosynthesis of nucleotide sugars,biosynthesis of amino acids,and pentose phosphate pathway;the pathways related to oxidative stress and antioxidant include glutathione metabolism,purine metabolism,riboflavin metabolism,retinol metabolism,tryptophan metabolism,flavonoid biosynthesis,biosynthesis of phenylpropanoids;Other pathways with significant abnormalities,insulin resistance,styrene degradation,prions disease,etc.These pathways provide a pavement for future studies on the bi-directional mechanism of action of type 2 diabetes mellitus with chronic periodontitis.