| Objectives:The aims of this study were to investigate the association of inflammation and oxidative stress of epicardial adipose tissue(EAT)with coronary artery disease(CAD),and he correlations of the levels of inflammatory adipocytokines including tumor necrosis factor-α(TNF-α),monocytic chemotecmic peptide-1(MCP-1),visfatin(VF),chemerin(CHEM),glutathione peroxidase-3(GPX-3),inductive nitric oxide synthase(i NOS),Angiotensinogen(AGT),adiponectin(ADP),interleukin-10(IL-10),endothelin-1(ET-1),apelin(AP LN),endothelial nitric oxide synthase(e NOS),transforming growth factor(TGF-β),with the prevalence and clinical risk factors of CAD.Methods:We consecutively enrolled 44 eligible inpatients in the Division of Cardiovascular Surgery from March 2019 to September 2019.Then,they were divided into CAD group undergone coronary artery bypass grafting group(n=26,60.96±8.88years),and NCAD(non-coronary artery disease)group undergone valves replacement(n=18,59.44±5.57years).The plasma levels of TNF-α,MCP-1,VF,CHEM,AGT,ET-1,i NOS,ADP,IL-10,TGF-β,APLN,GPX-3,e NOS were measured by enzyme-linked immunosorbent assay(ELISA).The protein expression of i NOS and e NOS in EAT were measured by Western Blotting.Hematoxylin-eosin staining(HE staining)was used to compare the adipocytes morphology and angiogenesis between EAT and SAT.Results:1 Clinical features: The prevalences of chest pain,hypertension and dyslipidemia were higher in the CAD group than those in the NCAD group[76.9(20/26)vs 11.1%(2/18),61.5%(16/26)vs 27.8%(5 / 18),78.3(18/26)vs 47.1(8/18),all P < 0.05].2 The levels of inflammatory factors:2.1 The plasma levels of pro-inflammatory factors via ELISA showed that TNF-α,MCP-1,VF and CHEM were significantly higher in CAD group than NCAD group,133.60(107.83,154.94)ng / L vs 102.30(85.11,110.45)ng / L,35.21(30.75,42.91)ng / L vs 27.86(22.87,30.00)ng / L,79.78(70.82,98.97)pg / m L vs 69.05(65.14,78.91)pg / m L,7.83(6.54,9.51)ng /L vs 6.73(6.13,7.65)ng/L,respectively(all P<0.05).2.2 The plasma levels of anti-inflammatory factors via ELISA were found that ADP,IL-10,TGF-β and APLN were significantly lower in the CAD group than the control group,321.10(282.75,348.35)pg / m L vs 361.60(353.75,395.25)pg / m L,92.07(79.41,95.06)ng / L vs 99.18(85.94,108.35)ng / L,1030.00(824.95,1183.50)pg / m L vs 1173.10(995.85,1613.50)pg / m L,922.00(832.00,995.5)ng / m L vs 963.10(931.80,1070.85)ng / m L,respectively(all P<0.05).2.3 The plasma level of oxidative stress,i NOS,was significantly higher in CAD group than that in NCAD group,3.29(3.03,3.62)umol/L vs 3.07(2.50,3.27)umol/L.However,the plasma concentrations of GPX-3 and e NOS were significantly lower in the CAD group than those in the NCAD group,3.29(3.03,3.62)pmol/m L vs 3.67(3.22,4.26)pmol/m L and 1.96(1.75,2.37)umol/L vs 2.62(1.95,2.88)umol/L,respectively(all P<0.05).2.4 The plasma levels of vasoconstriction relating factors by ELISA,AGT and ET-1,were significantly higher in CAD group than those in NCAD group,60.12(48.20,68.28)ng / m L vs 45.52(42.86,48.51)ng / m L,39.22(37.04,43.95)pg/m L vs 36.58(33.60,39.25)pg/m L,respectively(all P<0.05).2.5 EAT protein expression levels by Western Blotting showed that i NOS and the ratio of i NOS / e NOS were significantly higher in CAD group than in NCAD group: 1.09(0.85,1.25)vs 0.53(0.48,0.09),1.55(1.17,2.42)vs 0.55(0.48,0.78),respectively(all P <0.05).The e NOS was inversely lower in CAD group than that in NCAD group,0.66(0.41,0.85)vs 1.03(0.84,1.27),(P<0.05).3 Correlation between plasma inflammatory adipocytokine and clinical indicators3.1 Correlation between plasma inflammatory adipocytokinesPositive correlation between plasma inflammatory adipocytokines:AGT was positively correlated with TNF-α,MCP-1,VF,CHEM,Rs were0.472,0.310,0.561,0.473;TNF-α was positively correlated with MCP-1,VF,ET-1,i NOS,Rs were 0.573,0.533,0.313,0.416;MCP-1 was positively correlated with VF,Rs was 0.415;CHEM was positively correlated with ET-1,i NOS,Rs were 0.331,0.592;ET-1 was positively correlated with i NOS,Rs was 0.356;ADP was positively correlated with IL-10,APLN,GPX-3,e NOS,Rs were 0.561,0.358,0.432,0.432;IL-10 levels was positively correlated with TGF-β,APLN,Rs were 0.467,0.573;TGF-β was positively correlated with APLN,Rs was 0.531;APLN level was positively correlated with e NOS,Rs was 0.358;respectively(all P<0.05).Negative correlation between plasma inflammatory adipocytokines:MCP-1 was negatively correlated with ADP,IL-10,Rs were-0.374,-0.315;ET-1 was negatively correlated with IL-10,Rs was-0.306;respectively(all P<0.05).3.2 Correlation between plasma inflammatory adipocytokines and clinical factorsPlasma inflammatory adipocytokines are positively correlated with clinical characteristics:TNF-α was positively correlated with chest pain,CK,CK-MB,Tn-I,Rs were 0.366,0.436,0.371,0.385;MCP-1 was positively correlated with chest pain,Rs was 0.391;GPX-3 was positively correlated with atrial fibrillation and LA diameter,Rs were 0.295,0.509;ADP was positively correlated with atrial fibrillation,triglycerides,FT3,Rs were 0.406,0.320,0.357;IL-10 was positively correlated with low density lipoprotein,FT3,Rs were 0.347,0.603;ET-1 was positively correlated with lymphocytes,Rs was 0.494;e NOS was positively correlated with FT3,Rs was 0.722,respectively(all P<0.05).Plasma inflammatory adipocytokines are negatively correlated with clinical characteristics:TNF-α was negatively correlated with total protein and albumin,Rs were-0.326,-0.413;MCP-1 was negatively correlated with LA diameter,Rs was-0.311;VF was negatively correlated with LA diameter and total protein,Rs were-0.311 and-3.42;CHEM was negatively correlated with diastolic blood pressure,LA diameter,total protein,uric acid,Rs were-0.333,-0.63,-0.381,-0.349;AGT was negatively correlated with LA diameter,Rs was-0.349;GPX-3 was negatively correlated with hypertension,neutrophils,FT4,Rs were-0.285,-0.332,-0.344;i NOS was negatively correlated with total protein,albumin,BUN,Rs were-0.467,-0.344,-0.345;ADP was negatively correlated with chest pain,hypertension,dyslipidemia,AST,Rs were-0.372,-0.346,-0.291,-0.425;IL-10 was negatively correlated with hypertension and AST,Rs were-0.270 and-0.421;TGF-β was negatively correlated with AST and CK,Rs were-0.350 and-0.413;ET-1 was negatively correlated with LAD,total protein,albumin,Rs were-0.397,-0.429,-0.364;e NOS was negatively correlated with AST,blood glucose,Cr,Rs were-0.391,-0.431,-0.327,respectively(all P<0.05).4 Diagnostic test results4.1 Plasma pro-inflammatory cytokines and CADThe area under the curve of TNF-α,MCP-1,VF,CHEM,ET-1,AGT,and iNOS concentrations are 0.845(95%CI:0.728-0.962),0.833(95%CI:0.709-0.956),0.763(95%CI:0.615-0.910),0.703(95%CI:0.542-0.863),0.708(95%CI:0.537-0.878),0.865(95%CI:0.748-0.982),0.697(95%CI:0.529-0.864),respectively,(all P<0.05).And the cutoffs value are 120.15ng/L,30.44 ng/L,69.31 pg/m L,7.59 ng/L,37.93 pg/m L,46.56 ng/m L,2.84umol/L.The sensitivity are 64.00%,80.00%,88.00%,60.00%,72.00%,92.00%,92.00%.The specificity are 93.75%,81.25%,52.95%,76.47%,70.59%,70.59%,47.06%.4.2 Plasma anti-inflammatory adipokines and CADThe area under the curve of ADP,IL-10,TGF-β,APLN,GPX-3,and e NOS concentrations for CAD patients are 0.874(95% CI: 0.770-0.978),0.722(95% CI: 0.565-0.880),0.705(95% CI: 0.538-0.872),0.708(95% CI: 0.552-0.865),0.712(95% CI:0.549-0.875),0.689(95% CI: 0.519-0.859),respectively(all P<0.05).The cutoffs value for best diagnosis are 346.10 pg / m L,95.67 ng / L,1276.50 pg / m L,934.55 ng / m L,3.85pmol/m L,2.43 umol / L.The sensitivity are 76.00%,88.00%,92.00%,60.00%,96.00%,80.00%.The specificity are 88.24%,58.82%,47.06%,76.47%,41.18%,64.71 %.5 EAT pathology hematoxylin-eosin stainingThe HE statistical results showed that the average area of epicardial adipocytes was smaller than the average area of subcutaneous adipocytes(2822.52 ±765.83μm2 vs 3954.48 ± 854.82μm2),(P<0.05).The number of blood vessels in epicardial adipose tissue was more than that in subcutaneous adipose tissue(3.90 ± 1.36 vs 2.26±1.00),(P<0.05).Conclusions1 Inflammatory and oxidative stress of EAT are related to the development of CAD,and thereby is one of the neoteric mechanism of CAD.2 The newly-found inflammatory adipokines,CHEM,VF,ET-1,AGT,i NOS,APLN,TGF-β,GPX-3,IL-10 and e NOS may be used as important biomarkers for clinical diagnosis of CAD.They proves to have almost the same diagnostic potency as the typical biomarkers,such as TNF-α,MCP-1 and ADP.3 The biomarkers of inflammatory and oxidative stress of EAT may be an upcoming target for the clinical treatment for CAD. |
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