Study On Serum Oxidized Lipid Metabolism Profile And Inflammation-related Factors In Patients With Coronary Heart Disease And Blood Stasis Syndrome

Background:Coronary Heart Disease(CHD)is also known as coronary atherosclerotic heart disease,that is,coronary artery vascular atherosclerosis(Atherosclerosis,AS)lesions cause vascular lumen stenosis or obstruction,and ultimately lead t o myocardial ischemia and lack of oxygen even necrotic.Injury response is th e mainstream pathogenesis theory of CHD,that is,endothelial injury and lipid accumulation lead to chronic inflammation in the arterial wall to form atheroscl erotic plaques.Immune inflammation plays a key role in the initiation,progress ion,plaque rupture,and thrombosis of AS.Most previous studies have focused on the effector cells of typical inflammation,such as monocytes/macrophages,neutrophils and T cells.However,in addition to typical inflammatory effector c ells,a large number of experimental and clinical research evidences show that effector cells of allergic inflammation play an important role in the pathogenesi s of coronary artery disease(CAD),and eosinophils(EOS)are participated in t he key effector cells.After activation,EOS release a variety of signal factors t hat play an important role in leukocyte recruitment,endothelial cell damage,pl atelet activation,and thrombosis.In 2020,the European Society of Cardiology(ESC)classified CHD into two categories:Acute Coronary Syndrome(ACS)an d Chronic Coronary Syndrome(CCS).CHD has now become the leading cause of death for cardiovascular diseases(CVD)patients worldwide.From 1990 to 2016,a systematic analysis report on the burden of 333 diseases in 195 countr ies and regions around the world pointed out that CAD is one of the three ma jor causes of global disability adjusted life years(DALYs).In 2017,CHD mor tality accounted for 42.6%of CVD deaths in the United States.The prevalence and mortality of CHD in my country have also been increasing year by year.The increase is more obvious in rural areas,and the mortality rate of men is higher than that of women.Therefore,the global prevention and control of CH D still brooks no delay.In Chinese medicine,"chest pain","heartache",and "j ue heartache" belong to the category of CHD.The clinical features are mainly episodic chest tightness,chest pain,shortness of breath,or even chest pain thr ough the back,back pain through the heart,or accompanied by Dying.The cl assic treatise on TCM "The Synopsis of the Golden Chamber·Chest Bi,Hear t Pain,Short Qi and Pulse Syndrome No.9" considers its pathogenesis to be"Yang Wei Yin Xian",and proposes many effective clinical treatment rules,suc h as Tong Yang and Xuan Bi method,Strengthening and strengthening the bo dy method and Wentong analgesic method,etc.In recent years,the analysis of the research literature on CHD syndromes in traditional Chinese medicine show s that blood stasis syndrome accounts for the highest proportion of all TCM s yndromes,and the proportion in all CHD patients is increasing year by year.T herefore,to explore the role of oxidized lipid metabolites and inflammation-rela ted factors in the occurrence and development of CHD and CHD blood stasis syndrome has important clinical and practical significance.Objective:To study the correlation of oxidized lipid metabolites and inflammation-rel ated factors with CHD and CHD blood stasis syndrome,and provide certain re ference basis for its participation in CHD and CHD blood stasis syndrome.Methods:A collection of 120 CHD patients who were admitted to the Department o f Cardiology and General Department of Guang’anmen Hospital of China Acad emy of Chinese Medical Sciences from June 2020 to December 2020 were inc luded in the CHD group,and the CHD to be included in accordance with the"Diagnostic Standards for Coronary Heart Disease with Blood Stasis Syndrome"patients were divided into blood stasis syndrome group and non-blood stasis s yndrome group.At the same time,30 cases of health examiners in the prevent ive health care department of Guang’anmen Hospital of China Academy of Chi nese Medical Sciences from June to December 2020 were collected and includ ed in the health group.Collect all the selected subjects’ venous whole blood s amples on the day of consultation or in the early morning of the next day un der fasting conditions,use RT-PCR to detect the expression level of CCL11 m RNA in peripheral blood mononuclear cells(PBMC),and use ELISA method t o determine CCL11,ECP,TM,LTB4 expression levels,using LC-MS/MS platf orm to detect and analyze oxidized lipid metabolites,observe the oxidized lipid s between the healthy group and the CHD group,and between the CHD blood stasis syndrome group and the non-blood stasis syndrome group Differences in metabolites and inflammation-related factors.The experimental data of inflam mation-related factors were analyzed using SPSS 26.0 statistical software,and P<0.05 was used as the basis for judging statistically significant differences.T he oxidized lipid metabolites are processed by orthogonal partial least squares discriminant analysis(OPLS-DA)in the original data for centralized processing,and the MetaboAnalystR package OPLSR.Anal function in the R software is used for analysis.The screening rule:select VIP ≥1 and fold The metabolite with change≥2 or fold change≤0.5 is the final differential metabolite.Results:1.Baseline comparison between healthy group and CHD group,CHD bloo d stasis syndrome group and non-blood stasis syndrome groupThis study included 30 healthy people,120 CHD patients,80 CHD patient s with blood stasis syndrome,and 40 patients with non-blood stasis syndrome.Statistical analysis showed that there was no statistical difference between the CHD group and the healthy group in gender,age,BMI,GLU,ALT,AST,sCr,etc.;the CHD blood stasis syndrome group and the non-blood stasis syndrom e group had no statistical differences in gender,age,BMI,There was no statis tical difference in GLU,ALT,AST,sCr,etc.2.Results of EOS numbers in the healthy group and the CHD groupCompared with the healthy group,the number of EOS[(0.13±0.08)×109/L VS(0.10±0.08)×109/L],P=0.04)in the CHD group increased,and the differenc e was statistically significant.3.Results of CCL11,ECP,TM,LTB4 expression levels in healthy group a nd CHD groupCompared with the healthy group,the CHD group had CCL11(113.16±55.25 VS 77.74±33.07,P=0.007),ECP(24.44±5.01 VS 14.66±4.15,P=0.000),TM(3.11±1.20 VS 2.37±1.14,P=0.012),LTB4(113.84±44.27 VS 70.54±16.15,P=0.000)expression increased,and the difference was statistically significant.4.Binary Logistic regression analysis of CCL11,ECP,TM,LTB4 in CHD patientsUsing single factor logistic regression analysis,CCL11(OR=1.026,95%CI:1.007-1.045,P=0.007),ECP(OR=1.786,95%CI:1.356-2.351,P=0.000),TM(OR=1.717,95%CI:1.108-2.662,P=0.016),LTB4(OR=1.048,95%CI:1.021-1.075,P=0.000)were significantly correlated with CHD.Multivariate logistic regression analysis indicated that CCL11(OR=1.059,95%CI:1.011-1.109,P=0.015)and ECP(OR=2.149,95%CI:1.197-3.857,P=0.010)were significantly related to CHD;TM(OR=1.96,95%CI:0.654-5.499,P=0.239),LTB4(OR=1.092,95%CI:0.993-1.200,P=0.071)have no significant co rrelation with CHD.5.Results of the expression level of CCL11 mRNA in the healthy group a nd the CHD groupCompared with the healthy group,the expression level of CCL11 mRNA(3.80±2.74 VS 1.48±0.95,P=0.110)in PBMC of the CHD group was significa ntly increased,and the difference was statistically significant.6.Binary Logistic Regression Analysis of CCL11 mRNA in CHD PatientsSingle factor Logistic regression analysis showed that CCL11 mRNA(OR=1.986,95%CI:1.045-3.775,P=0.036)was significantly correlated with CHD.7.ROC curve analysis of CCL11,ECP,TM,LTB4 in CHD patientsROC curve analysis results indicate that the Area of CCL11 is 0.7337,the Cut-off value is 62.2pg/ml,the Sensitivity is 95.65%,and the Specificity is 50%;the Area of ECP is 0.9362,the Cut-off value is 19.58ng/ml,and the Sens itivity is 79.66%,Specificity is 93.33%;TM’s Area is 0.6777,Cut-off value is 1.975ng/ml,Sensitivity is 80.43%,Specificity is.46.15%;LTB4’s Area is 0.8089,Cut-off value is 81.57pg/ml,Sensitivity is 73.33%and Specificity is 83.33%.8.Results of EOS number of CHD blood stasis syndrome group and non-blood stasis syndrome groupThere was no significant difference in the number of EOS between CHD blood stasis syndrome group and non-blood stasis syndrome group[(0.14±0.06)×109/L VS(0.10±0.02)×109/L],P=0.83).9.Results of the expression levels of CCL11,ECP,TM,LTB4 in the CHD blood stasis syndrome groupCompared with the non-blood stasis syndrome group,the CHD blood stasi s syndrome group had higher expression levels of TM(3.53±1.15 VS 2.77±1.16,P=0.030)and LTB4(139.64±41.74 VS 96.65±37.56,P=0.001)and the differ ence was statistically significant;the expression levels of CCL11(113.25±66.49 VS 113.08±45.15,P=0.992)and ECP(24.87±5.14 VS 24.16±4.97,P=0.603)b etween CHD blood stasis syndrome group and non-blood stasis syndrome grou p was not statistically significant.10.Binary Logistic Regression Analysis of CCL11,ECP,TM,LTB4 in CH D Patients with Blood Stasis SyndromeSingle factor logistic regression analysis found that TM(OR=1.778,95%C I:1.039-3.041,P=0.036),LTB4(OR=1.026,95%CI:1.009-1.043,P=0.003)wer e significantly related to CHD.CCL11(OR=1.000,95%CI:0.989-1.011,P=0.992),ECP(OR=1.029,95%CI:0.926-1.144,P=0.596)had no significant correlati on with CHD.Multivariate logistic regression analysis indicated that CCL11(OR=0.987,95%CI:0.966-1.009,P=0.254),ECP(OR=1.067,95%CI:0.883-1.290,P=0.502),TM(OR=2.085,95%CI:0.742-5.860,P=0.164),LTB4(OR=1.013,95%CI:0.988-1.038,P=0.308)had no significant correlation with CHD.11.Results of CCL11 mRNA expression level in CHD blood stasis syndrom e groupCompared with non-blood stasis syndrome,the expression level of CCL11 mRNA(3.93±2.62 VS 3.60±2.99,P=0.703)in PBMC had no significant differe nce.12.ROC curve analysis of TM and LTB4 in CHD patients with blood stas is syndromeROC curve analysis results indicate that the Area of TM is 0.6762,the C ut-off value is 2.915ng/ml,the Sensitivity is 64%,and the Specificity is 71.43%;the Area of LTB4 is 0.7726,the Cut-off value is 123.9pg/ml,and the Sen sitivity is 85.19%,Specificity is 66.67%.13.Analysis of KEGG enrichment of oxidized lipid difference metabolites between healthy group and CHD blood stasis syndrome groupThe metabolites of oxidized lipid difference between healthy group and C HD blood stasis syndrome group were enriched in linoleic acid metabolic path way and metabolic pathway.14.Analysis of KEGG enrichment of oxidized lipid difference metabolite b etween healthy group and CHD non-blood stasis syndrome groupThe differential metabolites between the healthy group and the CHD non-b lood stasis syndrome group were enriched in the arachidonic acid metabolic pat hway,the inflammatory mediator regulation pathway of the TRP channel,the metabolic pathway,the PPAR signaling pathway,and the linoleic acid metaboli c pathway.15.Analysis of KEGG enrichment of oxidized lipid difference metabolite between CHD blood stasis syndrome and non-blood stasis syndrome groupsDifferential metabolite enrichment of CHD blood stasis syndrome and non-blood stasis syndrome in arachidonic acid metabolic pathway.Conclusion:1.Serum CCL11,ECP,LTB4,TM expression levels are significantly corre lated with CHD.2.There is a significant correlation between the expression level of CCL11 mRNA in PBMC and CHD.3.Serum TM and LTB4 expression levels are significantly correlated with CHD blood stasis syndrome,while serum CCL11,ECP and CHD blood stasis syndrome have no significant correlation.4.The expression level of CCL11 mRNA in PBMC has no significant cor relation with CHD blood stasis syndrome.5.Different metabolites of healthy group and CHD blood stasis syndrome group are enriched in linoleic acid metabolic pathway and metabolic pathway.6.The different metabolites between the healthy group and the CHD non-blood stasis syndrome group are enriched in the arachidonic acid metabolic pat hway,the inflammatory mediator regulation pathway of the TRP channel,the metabolic pathway,the PPAR signaling pathway,and the linoleic acid metaboli c pathway.7.The differential metabolites of CHD blood stasis syndrome and non-blo od stasis syndrome are enriched in the arachidonic acid metabolic pathway.