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Supplementary Figure 1. Flowchart for the screening, enrolment and follow-up of the study population.

Supplementary tables

Supplementary Table S1. Effect of liraglutide vs. placebo on mean 24-h BP at four intervals of the day

Treatment effect Difference 95% CI p-value

Liraglutide Placebo 24-h blood pressure parameters

Morning systolic BP (7-12) -4.8 (11.5) -3.3 (15.6) -1.5 (17.5) -9.0 to 6.1 0.686

Afternoon systolic BP (13-18) -0.6 (12.9) -3.4 (17.6) 2.8 (25.7) -8.1 to 13.6 0.602

Evening systolic BP (19-24) 4.3 (15.3) -4.8 (11.9) 9.2 (19.1) 1.1 to 17.2 0.028

Late Night systolic BP (01-06) 0.5 (18.4) -1.0 (14.9) 1.5 (26.7) -10.0 to 13.0 0.790

Morning diastolic BP (7-12) 2.4 (8.3) -0.4 (12.5) 2.8 (16.0) -4.1 to 9.7 0.406

Afternoon diastolic BP (13-18) 2.1 (10.4) -2.4 (11.7) 4.4 (17.0) -2.8 to 11.6 0.215

Evening diastolic BP (19-24) 5.2 (9.3) -4.5 (9.2) 9.7 (13.8) 3.9 to 15.5 0.002

Late Night diastolic BP (01-06) 1.9 (8.5) 0.1 (10.3) 1.8 (12.9) -3.8 to 7.3 0.523

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Data are expressed as mean (SD) unless stated otherwise. BP, blood pressure

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Supplementary Table S2. Treatment Effect by Subgroup

Subgroup Measurement Treatment effect Differenc

e 95% CI p-value N

Liragluti

de Placebo Baseline visit 1

mean 24-h systolic BP

<133.9 mmHg Mean 24-h Systolic BP,

mmHg -2.1

(4.6) 1.0

(11.6) -3.1

(14.4) -12.2 to

6.1 0.477 12

≥ 133.9 mmHg Mean 24-h Systolic BP, mmHg

-0.7 (10.8)

-7.4

(9.2) 6.7 (13.5) -1.9 to

15.2 0.116 12

Baseline visit 1 mean 24-h diastolic BP

<77.7 mmHg Mean 24-h Diastolic BP, mmHg

-2.2 (4.5)

-1.4 (10.9)

-0.8 (13.5)

-9.4 to

7.8 0.842 12

≥ 77.7 mmHg Mean 24-h Diastolic BP,

mmHg 2.0 (8.4) -2.7

(8.2) 4.7 (13.0) -3.6 to

13.0 0.240 12

Data are expressed as mean (SD) unless stated otherwise. BP, blood pressure

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Supplementary Table S3. List of Total Adverse Events in Study.

Group Event Liraglutide

period Placebo period Washout period Abnormal blood

test Amylase increased 1 (1.1) 0 (0) 0 (0)

Abnormal blood

test Anemia 1 (1.1) 0 (0) 0 (0)

Abnormal blood

test Creatinine Increased 1 (1.1) 0 (0) 1 (7.7)

Abnormal blood

test PSA Elevated 1 (1.1) 1 (2.6) 0 (0)

Cardiac Atrial Fibrillation 0 (0) 1 (2.6) 1 (7.7)

Cardiac Non Cardiac Chest Pain 0 (0) 0 (0) 2 (15.4)

Cardiac Chest Pain Cardiac 2 (2.2) 3 (7.9) 1 (7.7)

Cardiac Dyspnea 1 (1.1) 1 (2.6) 0 (0)

Cardiac Myocardial Infarction 2 (2.2) 0 (0) 0 (0)

Cardiac Palpitations 1 (1.1) 1 (2.6) 0 (0)

Cardiac Sinus Tachycardia 2 (2.2) 0 (0) 0 (0)

Gastrointestinal Anorexia 12 (13.2) 1 (2.6) 0 (0)

Gastrointestinal Bloating 1 (1.1) 0 (0) 0 (0)

Gastrointestinal Constipation 5 (5.5) 2 (5.3) 0 (0)

Gastrointestinal Dehydration 0 (0) 1 (2.6) 0 (0)

Gastrointestinal Diarrhea 7 (7.7) 4 (10.5) 1 (7.7)

Gastrointestinal Gastroesophageal Reflux

Disease 8 (8.8) 1 (2.6) 0 (0)

Gastrointestinal Gastroenteritis 0 (0) 1 (2.6) 1 (7.7)

Gastrointestinal Nausea 18 (19.8) 3 (7.9) 0 (0)

Gastrointestinal Stomach Pain 1 (1.1) 2 (5.3) 0 (0)

Gastrointestinal Vomiting 4 (4.4) 0 (0) 0 (0)

Infection Chills 1 (1.1) 0 (0) 0 (0)

Infection Infection eye 0 (0) 1 (2.6) 0 (0)

Infection Infections Other Cold 1 (1.1) 3 (7.9) 1 (7.7)

Infection Sinusitis 2 (2.2) 1 (2.6) 0 (0)

Infection Skin Infection 0 (0) 0 (0) 2 (15.4)

Muscle Myalgia 0 (0) 1 (2.6) 1 (7.7)

Neurological Dizziness 5 (5.5) 3 (7.9) 2 (15.4)

Neurological Erectile Dysfunction 0 (0) 1 (2.6) 0 (0)

Neurological Headache 2 (2.2) 0 (0) 0 (0)

Neurological Insomnia 0 (0) 2 (5.3) 0 (0)

Neurological Libido Decreased 1 (1.1) 0 (0) 0 (0)

Neurological Concussion 1 (1.1) 0 (0) 0 (0)

Neurological Somnolence 2 (2.2) 0 (0) 0 (0)

Other Fat Biopsy Related

Complication 2 (2.2) 0 (0) 0 (0)

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Renal, urine Proteinuria 1 (1.1) 0 (0) 0 (0)

Renal, urine Discoloration of Urine 0 (0) 1 (2.6) 0 (0)

Skin Pruritus 2 (2.2) 0 (0) 0 (0)

Skin Maculopapular Rash 1 (1.1) 1 (2.6) 0 (0)

Vascular Allergic reaction 0 (0) 1 (2.6) 0 (0)

Vascular Hot flash 1 (1.1) 1 (2.6) 0 (0)

Vascular Vascular Disorders -

Hypotension 1 (1.1) 0 (0) 0 (0)

Total events 91 38 13

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Supplementary Table S4. Period, sequence and carry over effects for primary and secondary outcomes

Parameter

P-value for period effect

P-value for sequence effect

P-value for carry over effect

Weight 0.415 0.528 0.519

Waist 0.599 0.813 0.934

HbA1C 0.007 0.058 0.041

Resting Heart rate 0.729 0.452 0.426

24-h systolic BP 0.236 0.264 0.371

24-h diastolic BP 0.286 0.444 0.618

24-h HR 0.778 0.999 1.000

Data are expressed as p-values for period, sequence and carry over effects. Analysis of variance for 2x2 crossover study was used (pkcross command in Stata 13.1).

HbA1C, glycated hemoglobin; BP, blood pressure; HR, heart rate

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Supplementary Table S5. Main characteristics of the studies included in the meta-analysis Study Design Liraglutide

subcutaneo us dosage

Subjects Duration Age, years Mean BMI HbA1C, %

Scholten,

2014 Non-

randomized , no

placebo- control

Titrated to

1.8 mg n=31;

T2D 7 weeks Median

age 64 (range: 37- 78)

31.9±4.4 7.7±1.3 %

Meier, 2015 Randomize d, no- placebo control

1.2 mg arm;

1.8 mg arm 1.2 mg arm (n=44) and 1.8 mg arm (n=46);

T2D

8 weeks 1.2 mg arm:

61.4±7.9 1.8 mg arm:

62.6±9.4

1.2 mg arm:30.5±

4.0 1.8 mg arm:

31.2±4.3

1.2 mg arm:

7.8±0.8%

1.8 mg arm:

7.9±0.8%

Lovshin, 2015

Randomize d, placebo- controlled, cross-over

Titrated to 1.8 mg

n=17;

T2D

3 weeks Median age 62 years

29.5 7%

Present study

Randomize d, placebo- controlled, cross-over

Titrated to 1.8 mg

n=24;

T2D

12 weeks 62.5±7.2 30.9±3.9 6.5±0.5%

BMI, body mass index; HbA1C, glycated hemoglobin; T2D, type 2 diabetes

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