Association Of Coronary Artery Disease With Pericoronary Adipose Tissue And PCAT Sourced Visfatin In Eldly

Part I: Relationship between Aging ￡uid Volume of Epicardial Adipose Tissue and Pericoronary Adipose TissueObjective: To discuss the relationship between aging and volume of epicardial adipose tissue（EAT） and pericoronary adipose tissue（PCAT）.Methods: Eighty normal ranged BMI patients who performed coronary computed tomography angiography（CCTA） were divided into different groups by age: group 1（<35 years old, n=10） : group 2（35-44 years old, n=20）;group3（45-54 years old, n=20） : group 4（55-64 years old, n=20） : group 5（>65years old, N=10）. Volume and thickness of EAT was measured by CCTA, and then PCAT of LM-LAD, LCX, RCA can be calculated through those acquisitions.Results: EAT volume increases along with aging: group 5（211.53 土14. 03cm’） >group 4（176. 25 土 21.63cm’） >group 3（157.13 土 27.24cm’）>group2（141.15±26.93cm3）>group 1（107.86±14. 31cm’）（p<0.05）. EAT thickness in group 1 is significantly thinner than other groups（p<0. 05）. LM-LAD PCAT volume increases with aging as well: group 5（9.06+0. 54cra’）>group 4（8. 39土 0. 60cm3） >group 3（7.90±1. 70cmO>group 2（6.54±0.86cm3）>group 1（4.45土0. 60cm’）, p<0.01. RCA PCAT volume also increases with aging as well: group5（11.90±1. 07cm’）〉group 4（10.18±1.07cm3）>group 3（9. 33±1. 30cm3）〉group2（7. 34土 1. 48cm’） >group 1（6.15±1.56cm’）, p<0.01. LCX PCAT volume in group1 is obviously less than other 4 groups（p<0.05）. Related analysis reveals that aging is related with EAT volume, LM-LAD PCAT volume and RCA PCAT volume（r is 0.749, 0.694 and 0.709 respectively, p〈0.05）.Conclusion: Volume of EAT, LM-LAD and RCA PCAT augments with aging.Part II: Relationship between Coronary Artery Disease and Volume of EAT and PCAT in old peopleObjective: To investigate the relationship between coronary artery disease and volume of EAT and PCAT in old people.Methods: Sixty old people who were performed with CCTA and coronaryangiography were divided into different groups by severity: Control group（normal coronary artery, n=9）;CAS group（coronary atherosclerosis,n=22）; CAD group（coronary artery disease, n=29）. Volume and thickness of EAT was measured by CCTA, and then PCAT of LM—LAD,LCX, RCA can be calculated through those acquisitions.Results: There is no statistical difference in three groups, although the volume of EAT is increased with the development of coronary atherosclerosis. Thickness of EAT in CAD group is thicker than those in control group [（3.98士 1. 07 mm >3.32±0. 47ram）, p=0.03]. LM-LAD PCAT in CAD group increases more than that in control and CAS group [（11. 26 ± 1.84cm^>9.86±1.41cm3>8.93±1.10）cm3,p〈0.01,p=0.003]. RCA PCAT in CAD group increases more than that control group [（14,14±2.69cin^>ll. 28±1. 64cm^）,p=0. 003）.Conclusion: With the extent of atherosclerosis, the volume of LM-LAD and RCA PCAT is inceased in old people.Part III: Relationship between Coronary Artery Disease and Visfatin Expression of PCAT in old peopleObjective: To discuss the relationship between coronary artery disease and visfatin expression of PCAT in old people by comparing visfatin expression of PCAT between patients with and without coronary artery disease.Methods: 15 normal figured elderly patients who were performed with cardiac surgery, including 9 cases of coronary artery bypass surgery（CABG）, and 6 cases of congenital heart disease or valve replacement surgery. Subcutaneous adipose tissue of chest wall, pericardial adipose tissue, myocardial fat and PCAT were collected. Real-time PGR was used to detect visfatin gene transcript level in adipose tissue. Western blotting was used to detect visfatin protein expression level in adipose tissue. At the mean time, immunohistochemical stain was used to analyze visfatin expression in adipose.Results: Immunohistochemistry stain reveals that visfatin protein in PCAT is significantly higher than other parts which including subcutaneous adipose tissue of chest wall, pericardial adipose tissue and myocardial fat. Visfatin protein content of PCAT in patients who were suffered from coronary artery disease is obviously higher than those without coronary artery disease. Real-time PGR results showed that visfatin mRNA in subcutaneous adipose tissue of chest wall, pericardial adipose tissue,myocardial fat and PCAT as well. Among all the tissues above, visfatin mRNA expression in PCAT is the highest（p〈0. 01）. visfatin iriRNA in PCAT in coronary artery disease（CAD） patients is higher than those without CAD[（3. 22±0. 31 > 2. 31±0.17）,（p<0. 01） ]. This difference is also found in myocardial fat（p<0. 01） and subcutaneous adipose tissue（p<0. 05）. But there is no such obvious difference shown in pericardial adipose tissue（p>0.05）. Western blotting results showed that visfatin protein expression in PCAT is higher than subcutaneous adipose tissue of chest wall, pericardial adipose tissue and myocardial fat. PCAT visfatin protein expression in patients who were suffered from CAD is higher than those without CAD（p=0. 001）. This difference is also found in myocardial fat（p=0. 008） and subcutaneous adipose tissue（p=0. 006）. But there is no such obvious difference shown in pericardial adipose tissue（p=0. 37）.Conclusion: Visfatin mRNA and protein expression in PCAT are higher than those in subcutaneous adipose tissue of chest wall, pericardial adipose tissue and myocardial fat. Abnormal high of visfatin mRNA and protein expression in PCAT may be related to CAD.Part IV: The Role of Visfatin in Coronary Artery Endothelial Cell Vitality and VCAM-1 ExpressionObjective: To investigate time-dose effect of visfatin on coronary artery endothelial cell vitality, and time effect of visfatin on VCAM-1expression.Methods: Human coronary artery endothelial cells which were cultured invitro were applied different concentration of visfatin（Ong/ml, lOng/ml,50ng/ml, lOOng/ml,200ng/ml, 400ng/ml） for different duration. MTT colorimetry was used to observe impact of visfatin on coronary endothelial cell vitality. Western blotting was used to detect impact of visfatin on VCAM-1 protein expression.Results: Coronary artery endothelial cell vitality and OD490 increases constantly from Oh to 48 h in 0ng/ml-200ng/ml visfatin group, while increase range in 50ng/ml-200ng/ml visfatin group were slightly smaller in both 24 h and 48 h than that in lOng/ml group and the control group.Coronary artery endothelial cell vitality increases slowly in 400ng/ml visfatin group. And it appears a downward trend at 24 h and 48 h. VCAM-1protein expression reaches maximum when coronary artery endothelial cells are stimulated by 200ng/ml visfatin group for 24 h.Conclusion: Low and medium concentration of visfatin can enhance coronary artery endothelial cell vitality. This effect has time time dependencies.High concentration of visfatin shows suppression effect on coronary artery endothelial cell vitality. Coronary artery endothelial cells can be induced to express VCAM—1,and peak in 24 h.Part V: Mechanism of Visfatin on Coronary Endothelial Cell Dysfunction and its Intervention ApproachesObjective: To discuss the mechanism of visfatin on coronary artery endothelial cell dysfunction, and protection effect of visfatin inhibitor（FK866）, NF- KB inhibitor（BAYl1-7082） and atorvastatin.Methods; Human coronary artery endothelial cells which were cultured in vitro were divided into different groups by adding different medications:visfatin group（visfatin 200 ng/ml）; visfatin inhibitor group（FK86610nfflol+ visfatin 200ng/ml）: NF- K B inhibitor group（BAYl1-7082 50umol+visfatin 200ng/ml）: atorvastatin group（atorvastatinlOumol+visfatin 200ng/ral） : control group（regular broth） ? MTT detection method was used to detect coronary endothelial cell proliferation vitality.Scratch test was used to calculate the number of migrating cells.Colorimetry was used to detect reactive oxygen species content in the cells. ELISA was used to measure the content of IL₆ and IL-8. Real-time PGR was used to detect mRNA expression of NO, ICAM-l, VCAM-1, NF-KB and ERK in coronary artery endothelial cells. Western blotting was used to detect protein expression of NO, ICAM-l, VCAM-1, NF-KB, ERK and P-ERK in cells. Irnmunohistochemical stain was used to analyze P65-NF- K B expression status.Results: 1) Effects on coronary endothelial cell proliferation : Coronary endothelial cell proliferation vitality can be enhanced by visfatin,while it can be inhibited by visfatin inhibitor, NF- KB inhibitor and atorvastatin. Atorvastatin has the strongest inhibition effect.Inhibition effect of visfatin inhibitor is similar with that of NF- KB inhibitor. 2) Effects on mRNA expression of ROS, IL-6, IL-8, ICAM-l and VCAM-1 in coronary artery endothelial cells: Expression of ROS, IL-6 and IL-8 in coronary endothelial cells can be enhanced by visfatin, while it can be significantly inhibited by visfatin inhibitor, NF- KB inhibitor and atorvastatin. Visfatin can induce the expression of inflammatory cytokines in coronary artery endothelial cells. Among all of them,visfatin inhibitor has the strongest inhibition effect. 3) Effects on mRNA and protein expression of adhesion molecules（ICAM-l and VCAM-1） in coronary artery endothelial cells: mRNA and protein expression of adhesion molecules in coronary artery endothelial cells can be remarkably increased by visfatin, while it can be significantly inhibited by visfatin inhibitor, NF- KB inhibitor and atorvastatin. Among all of them,visfatin inhibitor has the strongest inhibition effect. 4) Effects on mRNA and protein expression of eNOS in coronary artery endothelial cells: mRNA expression of eNOS in coronary artery endothelial cells can be inhibited by visfatin, while this inhibition effect can be reversed by visfatin inhibitor, NF- KB inhibitor and atorvastatin. Among all of them, visfatin inhibitor has the strongest reversal effect, 5) Effects on NF-KB approach in coronary artery endothelial cells: Fluorescence intensity of P65-NFKB mRNA and protein expression in coronary artery endothelial cells can be increased by visfatin, while it can be inhibited by visfatin inhibitor,NF- KB inhibitor and atorvastatin. Among all of them, NF- KB inhibitor has the strongest inhibition effect. 6) Effects on ERK signal pathway in coronary artery endothelial cells: ERK mRNA expression and P-ERK protein expression in coronary artery endothelial cells can be enhanced by visfatin, while it can be inhibited by visfatin inhibitor, NF- KB inhibitor and atorvastatin. At the mean time, expression of ERK protein does not increase, 7) Effects on coronary artery endothelial cell migration: The number of migrating coronary artery endothelial cell can be obviously enhanced by visfatin, while it can be inhibited by visfatin inhibitor, NF— KB inhibitor and atorvastatin. Among all of them, visfatin inhibitor has the strongest inhibition effect.Conclusion: By activating ERK signal pathway, visfatin can activate NFK B pathway（the nuclear transcription factor） and inflammatory reaction in coronary artery endothelial cells. Therefore, visfatin damages coronary artery endothelial cells. Through inhibiting ERK signal pathway,NF-KB activation can be reduced by visfatin inhibitor, NF— KB inhibitor and atorvastatin. Thus inflammatory reaction and cell migration will be suppressed, and coronary artery endothelial cells can be protected.Meanwhile, coronary artery endothelial cell protection effect may performed by FK866 by different ways other than ERK/NF-K B approach.