Introduction
Methods
Clinical study
Study population
Validation cohort
Biochemical measurements
Clinical follow up for adverse events
In vitrostudy
Human EPC isolation and culture
Transfection of siRNA
Migration assay
Tube formation assay
Adhesion assay
Western blot analysis
Statistical analysis
Results
Clinical study
Baseline clinical characteristics of study cohort
Characteristics | All participants (n = 1071) | Plasma endocan (pg/mL) | p-value | ||
---|---|---|---|---|---|
Tertile 1 (< 446.5) (n = 356) | Tertile 2 (446.5–666.7) (n = 358) | Tertile 3 (> 666.7) (n = 357) | |||
Age (years) | 65.5 ± 12.0 | 61.1 ± 10.6 | 65.8 ± 11.7 | 69.4 ± 12.3 | < 0.001 |
Male, no. (%) | 913 (85.2) | 316 (88.8) | 307 (85.8) | 290 (81.2) | 0.017 |
Body mass index (kg/m2) | 26.3 ± 4.4 | 26.8 ± 3.6 | 26.5 ± 3.8 | 25.6 ± 5.6 | < 0.001 |
Hypertension, no. (%) | 698 (65.2) | 221 (62.1) | 232 (64.8) | 245 (68.6) | 0.183 |
Diabetes mellitus, no. (%) | 408 (38.1) | 132 (37.1) | 134 (37.4) | 142 (39.8) | 0.722 |
Current smoker, no. (%) | 603 (56.3) | 206 (57.9) | 201 (56.2) | 196 (54.9) | 0.726 |
Use of anti-platelet, no. (%) | 992 (92.6) | 334 (93.8) | 337 (94.1) | 321 (89.9) | 0.056 |
Use of anti-coagulant, no. (%) | 28 (2.6) | 3 (0.8) | 11 (3.1) | 14 (3.9) | 0.029 |
Use of ACEi/ARB, no. (%) | 667 (62.3) | 228 (64.0) | 227 (63.4%) | 212 (59.4) | 0.379 |
Use of statin, no. (%) | 746 (69.7) | 267 (75.0) | 252 (70.4) | 227 (63.6) | 0.004 |
Fasting glucose (mg/dL) | 121.3 ± 45.0 | 121.6 ± 41.7 | 123.9 ± 45.3 | 118.3 ± 42.5 | 0.247 |
eGFR (mL/min/1.73 m2) | 74.5 ± 31.4 | 81.4 ± 25.6 | 77.6 ± 36.2 | 64.5 ± 29.0 | < 0.001 |
WBC count (CUMM) | 7389 ± 2317 | 7420 ± 2207 | 7406 ± 2418 | 7339 ± 2326 | 0.888 |
Total cholesterol (mg/dL) | 163.1 ± 35.7 | 164.3 ± 34.9 | 164.4 ± 36.6 | 160.5 ± 35.5 | 0.256 |
Triglyceride (mg/dL) | 134.5 ± 83.7 | 145.6 ± 90.2 | 139.5 ± 87.1 | 118.4 ± 70.2 | < 0.001 |
LDL-c (mg/dL) | 94.1 ± 28.8 | 94.7 ± 28.3 | 94.7 ± 29.7 | 93.0 ± 28.4 | 0.667 |
HDL-c (mg/dL) | 41.9 ± 10.7 | 41.3 ± 10.2 | 41.5 ± 10.5 | 43.0 ± 11.3 | 0.078 |
hsCRP (mg/dL) | 0.371 ± 0.955 | 0.343 ± 0.948 | 0.301 ± 0.531 | 0.458 ± 1.211 | 0.147 |
NT-pro-BNP (pg/mL) | 392.4 ± 854.9 | 218.4 ± 292.4 | 309.3 ± 415.6 | 649.3 ± 1354.5 | < 0.001 |
TNF-α (pg/mL) | 4.56 ± 5.27 | 3.30 ± 3.37 | 5.05 ± 6.45 | 5.37 ± 5.45 | < 0.001 |
Endocan (pg/mL) | 656.2 ± 500.6 | 338.5 ± 78.3 | 547.7 ± 60.2 | 1081.7 ± 670.2 | < 0.001 |
Variables | Cardiovascular events | p-value | |
---|---|---|---|
No (n = 864) | Yes (n = 207) | ||
Age (years) | 65.8 ± 11.8 | 64.0 ± 13.1 | 0.065 |
Male, no. (%) | 739 (85.5) | 174 (84.1) | 0.591 |
Body mass index (kg/m2) | 26.3 ± 4.6 | 26.4 ± 3.9 | 0.727 |
Hypertension, no. (%) | 566 (65.5) | 132 (63.8) | 0.637 |
Diabetes mellitus, no. (%) | 321 (37.2) | 87 (42.0) | 0.194 |
Current smoker, no. (%) | 468 (54.2) | 135 (65.2) | 0.004 |
Use of anti-platelet, no. (%) | 797 (92.3) | 195 (94.2) | 0.333 |
Use of anti-coagulant, no. (%) | 23 (2.7) | 5 (2.4) | 0.842 |
Use of ACEi/ARB, no. (%) | 543 (62.9) | 124 (59.9) | 0.433 |
Use of statin, no. (%) | 602 (69.7) | 144 (69.6) | 0.975 |
Fasting glucose (mg/dL) | 119.0 ± 41.5 | 130.7 ± 56.6 | 0.005 |
eGFR (mL/min/1.73 m2) | 75.0 ± 28.7 | 72.4 ± 40.7 | 0.384 |
WBC count (CUMM) | 7308 ± 2264 | 7707 ± 2495 | 0.039 |
Total cholesterol (mg/dL) | 162.5 ± 34.7 | 165.5 ± 39.5 | 0.317 |
Triglyceride (mg/dL) | 131.2 ± 78.8 | 148.5 ± 100.6 | 0.022 |
LDL-C (mg/dL) | 93.4 ± 28.8 | 97.1 ± 28.6 | 0.097 |
HDL-C (mg/dL) | 42.4 ± 10.8 | 40.0 ± 9.9 | 0.003 |
hsCRP (mg/dL) | 0.317 ± 0.769 | 0.570 ± 1.430 | 0.029 |
NT-pro-BNP (pg/mL) | 347.9 ± 858.5 | 578.1 ± 815.7 | < 0.001 |
TNF-α (pg/mL) | 4.31 ± 5.17 | 5.47 ± 5.54 | 0.016 |
Endocan (pg/mL) | 619.6 ± 424.2 | 808.9 ± 720.3 | < 0.001 |
Clinical outcomes
Outcomes, no. (%) | Plasma endocan (pg/mL) | p-value | ||
---|---|---|---|---|
Tertile 1 (< 446.5) (n = 356) | Tertile 2 (446.5–666.7) (n = 358) | Tertile 3 (> 666.7) (n = 357) | ||
Total CV events | 49 (13.8) | 58 (16.2) | 100 (28.0) | < 0.001 |
Hard CV events | 6 (1.7) | 12 (3.4) | 24 (6.7) | 0.002 |
CV mortality | 0 (0.0) | 1 (0.3) | 11 (3.1) | < 0.0001 |
Non-fatal MI | 5 (1.4) | 9 (2.5) | 10 (2.8) | 0.413 |
Non-fatal stroke | 1 (0.3) | 2 (0.6) | 3 (0.8) | 0.606 |
Unstable angina | 41 (11.5) | 42 (11.7) | 49 (13.7) | 0.613 |
Heart failure | 2 (0.6) | 2 (0.6) | 15 (4.2) | < 0.001 |
PAD | 0 (0.0) | 2 (0.6) | 12 (3.4) | < 0.001 |
Plasma endocan was an independent predictor for CV events
Variables | Univariate | Multivariate* | ||
---|---|---|---|---|
HR [95% CI] | P value | HR [95% CI] | p-value | |
A. Hard cardiovascular events | ||||
Endocan (pg/mL) | 1.001 [1.000–1.001] | < 0.001 | 1.000 [1.000–1.001] | 0.062 |
Endocan tertile | 2.049 [1.351–3.1074] | < 0.001 | 1.774 [1.118–2.813] | 0.015 |
Age (years) | 1.004 [0.979–1.030] | 0.749 | ||
Current smoking | 1.257 [0.674–2.344] | 0.471 | ||
Fasting glucose (mg/dL) | 1.009 [1.005–1.014] | < 0.001 | 1.011 [1.005–1.017] | < 0.001 |
WBC(CUMM) | 1.000 [1.000–1.000] | 0.085 | 1.000 [1.000–1.000] | 0.609 |
Triglyceride (mg/dL) | 1.000 [0.996–1.004] | 0.996 | ||
HDL-c (mg/dL) | 0.987 [0.958–1.018] | 0.407 | ||
hsCRP (mg/dL) | 1.157 [0.982–1.364] | 0.082 | 1.072 [0.872–1.317] | 0.509 |
NT-proBNP (pg/mL) | 1.000 [1.000–1.000] | 0.006 | 1.000 [1.000–1.000] | 0.113 |
TNF-α (pg/mL) | 1.023 [0.975–1.073] | 0.349 | ||
B. Total cardiovascular events | ||||
Endocan (pg/mL) | 1.000 [1.000–1.001] | < 0.001 | 1.000 [1.000–1.001] | < 0.001 |
Endocan tertile | 1.555 [1.306–1.851] | < 0.001 | 1.797 [1.452–2.225] | < 0.001 |
Age (years) | 0.988 [0.977–0.999] | 0.040 | 0.978 [0.964–0.992] | 0.003 |
Current smoking | 1.514 [1.137–2.015] | 0.005 | 1.438 [1.022–2.024] | 0.037 |
Fasting glucose (mg/dL) | 1.005 [1.002–1.007] | < 0.001 | 1.003 [1.000–1.006] | 0.045 |
WBC(CUMM) | 1.000 [1.000–1.000] | 0.015 | 1.000 [1.000–1.000] | 0.468 |
Triglyceride (mg/dL) | 1.002 [1.001–1.003] | 0.007 | 1.001 [0.999–1.003] | 0.158 |
HDL-c (mg/dL) | 0.980 [0.966–0.994] | 0.004 | 0.992 [0.976–1.009] | 0.365 |
hsCRP (mg/dL) | 1.170 [1.070–1.280] | < 0.001 | 1.083 [0.958–1.224] | 0.204 |
NT-proBNP (pg/mL) | 1.000 [1.000–1.000] | 0.001 | 1.000 [1.000–1.000] | 0.187 |
TNF-α (pg/mL) | 1.029 [1.007–1.052] | 0.010 | 1.013 [0.989–1.038] | 0.289 |