| Background:Coronary atherosclerosis heart disease refers to the stenosis of luminal due to the atherosclerosis,cramp or block of coronary artery,which can lead to ischemia,hypoxia or necrosis of myocardial caused by heart disease.Generally,it is termed coronary heart disease(CHD),or coronary artery disease for short.As we all know,it is the most common type of atherosclerosis that leads to organ lesion.Because the completely blocked of coronary artery is often caused by thrombosis,it is also known as coronary atherothrombotic heart disease in recent years.Coronary heart disease tends to occur in the elderly over the age of 40 and the morbidity in male is higher than female.Meanwhile,coronary heart disease is widespread in brain workers and developed countries,which has became the most common disease of heart disease in Occident.According to the data of WHO published in 2006,there are 7.2 million people died for coronary heart disease across the globe every year.As the worldwide aging of population,the number of the mortality of coronary heart disease will sustain to rise,and it is estimated that the death toll of patients with coronary heart disease will increase to 11 million by 2020.Mounts of researches suggest that type 2 diabetes mellius(T2DM)has became a equally dangerous disease in cardiovasclar system as well as coronary heart disease.Remarkably,the cardiovascular complications of T2 DM is one of the most common reason which can lead to death in patients with T2 DM.A large proportion of coronary heart disease are accompanied with type 2 diabetes mellius.Plenty of studies have shown that both coronary heart disease and type 2 diabetes mellius are chronic inflammatory diseases which behave as metabolic disorders and immune abnormalities.According to the recent researches,inflammasome plays an important role in the process of development of all kinds of chronic inflammatory diseases.Notably,nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome might be significantly related to the occurrence of coronary heart disease in patients with type 2 diabetes.In conclusion,our study aims at comparing the mRNA transcription information and protein expression level of NLRP3,apoptosis-associated speck-like protein(ASC),Caspase-1 in peripheral blood mononuclear cells by collecting different cases,as well as the levels of interleukin-1?(IL-1?),interleukin-18(IL-18)in plasma.In order to explore the role and mechanism of nucleotide binding oligomerization domain-like receptor protein 3 inflammasome in the development and progression of type 2 diabetes complicated with coronary heart disease.Methods:1.Cases collection and grouping: 110 patients who hospitalized in the department of Cardiology in our hospital from March to August in 2016 were enrolled.They were divided into 4 groups,including normal individuals(n=30),patients with CHD(n=30),patients with T2DM(n=20)and patients with T2 DM complicated with CHD(n=30).Meanwhile,the general clinical data and biochemical indicators were collected.2.Isolated peripheral blood mononuclear cells and plasma:20ml of fresh venous blood was collected from each patient into a EDTA anticogulated tube respectively.Peripheral blood mononuclear cells(PBMCs)were isolated from each patient by Ficoll-Hypaque density gradient centrifugation,and the plasma of each patient was isolated by high speed centrifugation as well.3.The mRNA transcription information of NLRP3,ASC,Caspase-1in PBMCs: the mRNA transcription information of NLRP3,ASC,Caspase-1in PBMCs were detected by real-time quantitative polymerase chain reaction(RT-qPCR).4.The protein expression of NLRP3,ASC,Caspase-1in PBMCs: the protein expression level of NLRP3,ASC,Caspase-1in PBMCs were detected by Western blot.5.The levels of IL-1?,IL-18 in plasma: the levels of IL-1?,IL-18 in plasma were measured by ELISA.6.The differences and correlations of the above laboratory indexs were tested to confirm the role of NLRP3 inflammasome in occurrence of type 2 diabetes complicated with coronary heart disease.Results:1.The clinical biochemical data:there was no significantly difference in gender,age,BMI and smoking history among groups(P>0.05).Otherwise,no significantly differences were detected in serum among total cholesterol(TC),triglyceride(TG),hypersensitive C-reactive protein(hs-CRP)and uric acid(P>0.05).2.The results of RT-qPCR:the mRNA transcription levels of NLRP3,ASC,Caspase-1in PBMCs of patients with CHD,patients with T2 DM and patients withT2 DM complicated with CHD were obviously elevated compared with the healthy individuals(P<0.05),but we did not obtain significantly differences among patients with CHD,patients with T2 DM and patients withT2 DM complicated with CHD(P>0.05).3.The results of Western blot: the protein expression levels of NLRP3,ASC,Caspase-1in PBMCs of patients with CHD,patients with T2 DM and patients withT2 DM complicated with CHD were remarkably elevated compared with the normal control(P<0.05).The NLRP3 protein expression levels in patients with T2 DM complicated with CHD were significantly higher than patients with CHD and patients with T2DM(P<0.05),while the ASC,Caspase-1protein expression levels were no significantly differences in patients with CHD,patients with T2 DM and patients withT2 DM complicated with CHD(P>0.05).4.The results of ELISA:1)The levels of IL-1? in plasma were(23.35±0.81,CHD)ng/L,(22.86±1.67,T2DM)ng/L,(26.78±1.97,T2 DM complicated with CHD)ng/L,which were significantly higher than(12.53±2.79,control)ng/L(P<0.05).At the same time,the levels of IL-1? in patients withT2 DM complicated with CHD were significantly higher than patients with CHD and patients with T2DM(P<0.05).2)2The levels of IL-18 in plasma were(100.00±12.90,CHD)ng/L,(116.21±11.77,T2DM)ng/L,(138.49±16.36,T2 DM complicated with CHD)ng/L,which were significantly higher than(50.04±10.20,control)ng/L(P<0.05).At the same time,the levels of IL-18 in patients withT2 DM complicated with CHD were significantly higher than patients with CHD and patients with T2DM(P<0.05).5.The results of correlation analysis:1)The levels of IL-1? in plasma were positively correlated with the mRNA transcription levels of NLRP3(r=0.278,p=0.002),ASC(r=0.461,p=0.000),Caspase-1(r=0.262,p=0.004)in PBMCs,as well as the protein expression levels of NLRP3(r=0.591,p=0.000),ASC(r=0.354,p=0.001),Caspase-1(r=0.450,p=0.000)in PBMCs respectively.2)The levels of IL-18 in plasma were positively correlated with the mRNA transcription levels of ASC(r=0.386,p=0.000),Caspase-1(r=0.295,p=0.001)in PBMCs,as well as the protein expression levels of NLRP3(r=0.581,p=0.000),ASC(r=0.404,p=0.000),Caspase-1(r=0.442,p=0.000)in PBMCs respectively.However,There was no significant correlation between the levels of IL-18 in plasma and the mRNA transcription levels of NLRP3 in PBMCs(r=0.168,p=0.067).Conclusion:1.The aberrant activation of NLRP3 inflammasome may play an important role in occurrence of type 2 diabetes complicated with coronary heart disease by the NLRP3 inflammasome-proinflammatory cytokine(IL-1?)axis.2.The protein expression levels of NLRP3 were remarkably elevated in patients with T2 DM complicated with CHD.Our results suggest that NLRP3 protein,a speed limiting element in the activation of NLRP3 inflammasome,may play a critical role in this process.Therefore,the abnormal accumulation of NLRP3 protein may be the key of NLRP3 inflammasome activition and the occurence of inflammatory disease. |
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