J Clin Med Res

Journal of Clinical Medicine Research, ISSN 1918-3003 print, 1918-3011 online, Open Access

Article copyright, the authors; Journal compilation copyright, J Clin Med Res and Elmer Press Inc

Journal website http://www.jocmr.org

Original Article

Volume 11, Number 12, December 2019, pages 789-797

Plasma Levels of Myocardial MicroRNA-133a Increase by Intraoperative Cytokine Hemoadsorption in the Complex Cardiovascular Operation

Figure 2. Changes of myogenic miRNAs after surgery. Con: control group; HA: hemoadsorption group. Time points: T1: after induction of anesthesia, T2: 3 h (after skin incision), T3: 6 h (after ICU arrival), T4: 18 h (the first postoperative day, 5 o’clock a.m.) and T5: 42 (the second postoperative day, 5 o’clock a.m.) after aortic declamping. Label a: The plasma levels of miRNA-133a were increased postoperatively in both groups but increased much more in the HA group at T2 (Con 2 versus HA 2) and T4 (Con 4 versus HA 4). Label b: The plasma levels of miRNA-1 were increased postoperatively in both groups, but without the difference between groups.

**Table 1.** Characteristics of the Two Groups
	HA (n = 15)	Control (n = 13)	P-value
Values are presented as the number (n), percentage (%) or median (min.; max.). P values indicating statistical significance were calculated using a t-test for continuous variables and a Mann-Whitney U-test for categorical variables. HA: hemoadsorption; BMI: body mass index; NYHA: New York Heart Association classification of heart failure; Euroscore: classification of early (1 month) mortality; LVEF: left ventricle ejection fraction; ACE: angiotensin-converting-enzyme. Bypass time: duration of the extracorporeal circulation; Aortic clamp time: duration of cardioplegic ischemic arrest.
Preoperative characteristics
Age (years)	50 ± 10	54 ± 15	0.526
Female	2	0	0.206
Male	13	13	0.26
BMI	30.5 ± 3.6	27.7 ± 3.8	0.065
NYHA classification	1 (1; 2)	2 (1; 2)	0.956
Euroscore II.	4.2 ± 2.1	4.1 ± 1.9	0.907
Previous myocardial infarction	0	2	0.484
Peripheral venous disease	1	0	0.464
Hypertension	3	2	0.491
Diabetes	1	4	0.333
Hypercholesterolemia	4	3	0.498
LVEF total (%)	52.9 ± 13.9	58.6 ± 7.2	0.218
30-40%	1	1	1
40-50%	2	0	0.206
More than 50%	10	14	0.153
Drug history
ACE inhibitors	4	7	0.46
Beta-blockers	3	2	0.491
Statins	5	6	1
Insulin	0	1	1
Sulphonylurea	1	2	1
Intraoperative characteristics
Ross operation (abs/%)	14 (93)	13 (93)	1
David operation (abs/%)	1 (7)	1 (7)	1
Bypass time (min)	198 ± 15	182 ± 44	0.69
Aortic clamp time (min)	132 ± 18	134 ± 25	0.846

Table 1. Characteristics of the Two Groups

HA (n = 15)

Control (n = 13)

P-value

Values are presented as the number (n), percentage (%) or median (min.; max.). P values indicating statistical significance were calculated using a t-test for continuous variables and a Mann-Whitney U-test for categorical variables. HA: hemoadsorption; BMI: body mass index; NYHA: New York Heart Association classification of heart failure; Euroscore: classification of early (1 month) mortality; LVEF: left ventricle ejection fraction; ACE: angiotensin-converting-enzyme. Bypass time: duration of the extracorporeal circulation; Aortic clamp time: duration of cardioplegic ischemic arrest.

Preoperative characteristics

Age (years)

50 ± 10

54 ± 15

0.526

Female

0.206

Male

0.26

BMI

30.5 ± 3.6

27.7 ± 3.8

0.065

NYHA classification

1 (1; 2)

2 (1; 2)

0.956

Euroscore II.

4.2 ± 2.1

4.1 ± 1.9

0.907

Previous myocardial infarction

0.484

Peripheral venous disease

0.464

Hypertension

0.491

Diabetes

0.333

Hypercholesterolemia

0.498

LVEF total (%)

52.9 ± 13.9

58.6 ± 7.2

0.218

30-40%

40-50%

0.206

More than 50%

0.153

Drug history

ACE inhibitors

0.46

Beta-blockers

0.491

Statins

Insulin

Sulphonylurea

Intraoperative characteristics

Ross operation (abs/%)

14 (93)

13 (93)

David operation (abs/%)

1 (7)

Bypass time (min)

198 ± 15

182 ± 44

0.69

Aortic clamp time (min)

132 ± 18

134 ± 25

0.846

**Table 2.** MiRNA Quantification
Group	T	miRNA-23b right atrium	P1	P2	miRNA-1 right atrium	P1	P2	miRNA-1 blood	P1	P2	miRNA-133a blood	P1	P2	miRNA-223 blood	P1	P2	miRNA-126 blood	P1	P2
The data are presented as median (min.; max). P1: Comparisons of continuous parameters between groups were computed using a Mann-Whitney U-test. P2: A Wilcoxon paired test was used to assess the significance of time related changes within groups. Plasma and tissue miRNA levels were presented as the fold-change relative to T1 (preoperative value), defined as 1,0. Statistically significant differences are indicated by an asterisk (*) between groups and a hash tag (#) within groups. HA: hemoadsorption group. T1: after induction of anesthesia; T2: 3 h; T3: 6 h; T4: 18 h; T5: 42 h after myocardial reperfusion.
HA	1	1			1			1			1	1	1	1			1
	2	1.081 (0.437; 14.432)	0.243	0.422	1.126 (0.824; 1.821)	0.821	0.075	24.022 (0.374; 625)	0.50	0.013#	63.135 (0.684; 790.322)	0.037*	0.004#	0.684 (0.298; 10.491)	0.271	0.552	0.925 (0.446; 23.136)	0.077	0.65
	3							16.105 (0.723; 9,753)	0.451	0.004#	38.315 (1.338; 1,024)	0.135	0.003#	1.25 (0.375; 47.266)	0.105	0.239	0.951 (0.468; 35.055)	0.316	0.308
	4							9.413 (0.116; 6,323)	0.369	0.013#	62.938 (0.783; 1,008)	0.017*	0.004#	2.143 (0.561; 10.337)	0.57	0.005#	0.727 (0.269; 4.018)	0.443	0.875
	5							4.265 (0.08; 1,148)	0.414	0.116	5.215 (0.014; 110.887)	0.214	0.028#	1.987 (0.555; 9.282)	0.67	0.01#	0.631 (0.006; 3.298)	0.877	0.182
Control	1	1			1			1			1	1	1	1			1
	2	0.778 (0.144; 2.889)		0.888	1.180 (0.737; 2.683)		116	45.87 (1.926; 39,471)		0.012#	2.24 (0.408; 71.073)		0.036#	0.645 (0.194; 1.265)		0.036#	0.435 (0.263; 2.911)		0.214
	3							101.264 (1.203; 905,424)		0.018#	13.681 (1.234; 77.345)		0.018#	0.561 (0.140; 1.725)		0.398	0.62 (0.186; 2.287)		0.208
	4							24.759 (0.802; 1,041,863)		0.017#	3.786 (0.366; 62.938)		0.036#	2.082 (0.728; 4.415)		0.036#	0.551 (0.169; 2.949)		0.327
	5							7.412 (0.08; 1,148)		0.049#	0.811 (0.174; 6.980)		0.779	3.16 (0.430; 16.599)		0.025#	0.619 (0.270; 6.137)		0.208

Table 2. MiRNA Quantification

Group

miRNA-23b right atrium

miRNA-1 right atrium

miRNA-1 blood

miRNA-133a blood

miRNA-223 blood

miRNA-126 blood

The data are presented as median (min.; max). P1: Comparisons of continuous parameters between groups were computed using a Mann-Whitney U-test. P2: A Wilcoxon paired test was used to assess the significance of time related changes within groups. Plasma and tissue miRNA levels were presented as the fold-change relative to T1 (preoperative value), defined as 1,0. Statistically significant differences are indicated by an asterisk (*) between groups and a hash tag (#) within groups. HA: hemoadsorption group. T1: after induction of anesthesia; T2: 3 h; T3: 6 h; T4: 18 h; T5: 42 h after myocardial reperfusion.

1.081 (0.437; 14.432)

0.243

0.422

1.126 (0.824; 1.821)

0.821

0.075

24.022 (0.374; 625)

0.50

0.013#

63.135 (0.684; 790.322)

0.037*

0.004#

0.684 (0.298; 10.491)

0.271

0.552

0.925 (0.446; 23.136)

0.077

0.65

16.105 (0.723; 9,753)

0.451

0.004#

38.315 (1.338; 1,024)

0.135

0.003#

1.25 (0.375; 47.266)

0.105

0.239

0.951 (0.468; 35.055)

0.316

0.308

9.413 (0.116; 6,323)

0.369

0.013#

62.938 (0.783; 1,008)

0.017*

0.004#

2.143 (0.561; 10.337)

0.57

0.005#

0.727 (0.269; 4.018)

0.443

0.875

4.265 (0.08; 1,148)

0.414

0.116

5.215 (0.014; 110.887)

0.214

0.028#

1.987 (0.555; 9.282)

0.67

0.01#

0.631 (0.006; 3.298)

0.877

0.182

Control

0.778 (0.144; 2.889)

0.888

1.180 (0.737; 2.683)

116

45.87 (1.926; 39,471)

0.012#

2.24 (0.408; 71.073)

0.036#

0.645 (0.194; 1.265)

0.036#

0.435 (0.263; 2.911)

0.214

101.264 (1.203; 905,424)

0.018#

13.681 (1.234; 77.345)

0.018#

0.561 (0.140; 1.725)

0.398

0.62 (0.186; 2.287)

0.208

24.759 (0.802; 1,041,863)

0.017#

3.786 (0.366; 62.938)

0.036#

2.082 (0.728; 4.415)

0.036#

0.551 (0.169; 2.949)

0.327

7.412 (0.08; 1,148)

0.049#

0.811 (0.174; 6.980)

0.779

3.16 (0.430; 16.599)

0.025#

0.619 (0.270; 6.137)

0.208

**Table 3.** Postoperative Characteristics
Postoperative characteristics	HA (n = 15)	Control (n = 13)	P-value
Values are presented as number (abs), percentage (%), median (min.; max). 3-1 dif: the difference between T3 and T1. Laboratory data are compared between the groups as the difference between the peek value (T3) and the preoperative value (T1). The listed P values of statistical tests were calculated by using Mann-Whitney U-test. HA: hemoadsorption; Neutrophil-SFL (septic marker): the fluorescent light intensity of the neutrophil area on the white blood cell differential scattergram; CRP: C-reactive protein; ICU: intensive care unit; MoCA: Montreal cognitive assessement; PCT: procalcitonin.
Platelets, g/L (3-1 diff)	-31 (-21 - 60)	-29 (-74 - 87)	0.983
Leukocytes, g/L (3-1 diff)	6.7 (2 - 14.9)	3.6 (1.5 - 13.9)	0.34
Neutrophils, g/L (3-1 diff)	4.6 (2.5 - 13.8)	4 (1.9 - 14.3)	0.389
Immature granulocyt, g/L (3-1 diff)	0.04 (0 - 0.10)	0.03 (0.01 - 0.13)	0.576
Neutrophil-SFL, ch (3-1 dif)	4.6 (2.3 - 7.2)	5.5 (-25 - 7)	0.389
Free Hb, g/L (2-1 dif)	383 (181 - 463)	369 (172 - 477)	0.599
C-reactive protein, mg/L (3-1 dif)	163 (41 - 326)	169 (92 - 251)	0.747
PCT, µg/L (3-1 dif)	0.3 (0.1 - 1)	0.63 (0.1 - 1.1)	0.062
Fibrinogen, g/L (3-1 dif)	0.9 (0.4 - 1.9)	1 (0.2 - 1.8)	0.631
Blood loss (mL)	420 (260; 1,200)	440 (280; 1,330)	0.504
Extubation (h)	4 (2; 10)	5 (3; 8)	0.176
Length of stay in ICU (days)	2 (1; 3)	3 (1; 4)	0.085
Hospitalization time (days)	9 (7; 14)	8 (7; 15)	0.419
MoCA (preoperation - discharge dif.)	1 (-2; 5)	1 (-3; 5)	0.817
Postoperation delirium (abs./%)	0	2 (14)	0.224
Atrial fibrilation (abs/%)	4 (27)	3 (20)	0.682
Vasopresors (abs/%)	4 (27)	3 (20)	0.682
Inotropics (abs/%)	4 (27)	4 (29)	0.977
Wound infection (abs/%)	1 (7)	2 (14)	0.598
Mortality (3 months)	0	0

Table 3. Postoperative Characteristics

Postoperative characteristics

HA (n = 15)

Control (n = 13)

P-value

Values are presented as number (abs), percentage (%), median (min.; max). 3-1 dif: the difference between T3 and T1. Laboratory data are compared between the groups as the difference between the peek value (T3) and the preoperative value (T1). The listed P values of statistical tests were calculated by using Mann-Whitney U-test. HA: hemoadsorption; Neutrophil-SFL (septic marker): the fluorescent light intensity of the neutrophil area on the white blood cell differential scattergram; CRP: C-reactive protein; ICU: intensive care unit; MoCA: Montreal cognitive assessement; PCT: procalcitonin.

Platelets, g/L (3-1 diff)

-31 (-21 - 60)

-29 (-74 - 87)

0.983

Leukocytes, g/L (3-1 diff)

6.7 (2 - 14.9)

3.6 (1.5 - 13.9)

0.34

Neutrophils, g/L (3-1 diff)

4.6 (2.5 - 13.8)

4 (1.9 - 14.3)

0.389

Immature granulocyt, g/L (3-1 diff)

0.04 (0 - 0.10)

0.03 (0.01 - 0.13)

0.576

Neutrophil-SFL, ch (3-1 dif)

4.6 (2.3 - 7.2)

5.5 (-25 - 7)

0.389

Free Hb, g/L (2-1 dif)

383 (181 - 463)

369 (172 - 477)

0.599

C-reactive protein, mg/L (3-1 dif)

163 (41 - 326)

169 (92 - 251)

0.747

PCT, µg/L (3-1 dif)

0.3 (0.1 - 1)

0.63 (0.1 - 1.1)

0.062

Fibrinogen, g/L (3-1 dif)

0.9 (0.4 - 1.9)

1 (0.2 - 1.8)

0.631

Blood loss (mL)

420 (260; 1,200)

440 (280; 1,330)

0.504

Extubation (h)

4 (2; 10)

5 (3; 8)

0.176

Length of stay in ICU (days)

2 (1; 3)

3 (1; 4)

0.085

Hospitalization time (days)

9 (7; 14)

8 (7; 15)

0.419

MoCA (preoperation - discharge dif.)

1 (-2; 5)

1 (-3; 5)

0.817

Postoperation delirium (abs./%)

2 (14)

0.224

Atrial fibrilation (abs/%)

4 (27)

3 (20)

0.682

Vasopresors (abs/%)

4 (27)

3 (20)

0.682

Inotropics (abs/%)

4 (27)

4 (29)

0.977

Wound infection (abs/%)

1 (7)

2 (14)

0.598

Mortality (3 months)