inflammatory markers and anti- inflammatory effects of insulin
TRANSCRIPT
Inflammatory Markers and AntiInflammatory Markers and Anti-- Inflammatory Effects of InsulinInflammatory Effects of Insulin
Paresh Dandona, BSc, MD, DPhil, FRCP, FACP, FACC, FACE
Distinguished Professor of Medicine and Pharmacology School of Medicine and Biomedical SciencesDivision Head, Endocrinology and Metabolism
State University of New York at BuffaloFounder and Medical Director,
Diabetes-Endocrinology Center of Western New York Head, Division of Endocrinology
Kaleida Health/Millard Fillmore Hospital, Buffalo, New York
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2
Cum
ula
tive
Rat
e of Rec
urr
ent
MI
or
Coro
nar
y D
eath
Lowering CRP Lowering CRP with Statinwith Statin
Therapy is BeneficialTherapy is Beneficial
Follow-up (Years)
0.0 0.5 1.0 1.5 2.0 2.50
1
2
3
4
5
6
% C
V E
vents
at
8 Y
ears
Higher CRP Levels Higher CRP Levels are Associated with are Associated with
Adverse EventsAdverse Events
Adapted from Ridker PM et al. Circulation 2003;107:391-397. | Ridker PM et al. N Engl J Med 2005;352:20-28. Copyright Massachusetts Medical Society. All rights reserved.
CRP <1CRP 1–3
CRP >3
CRP 2 mg/L
CRP <2 mg/L
0.00
0.02
0.04
0.06
0.08
0.10
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3Clinical Relevance of Achieved LDLClinical Relevance of Achieved LDL--C and Achieved CRP Combined after Treatment C and Achieved CRP Combined after Treatment
with Statin Therapy: with Statin Therapy: PROVE ITPROVE IT––TIMI 22TIMI 22Rec
urr
ent
MI
or
Coro
nar
y D
eath
(%)
Follow-up (Years)0.0 0.5 1.0 1.5 2.0 2.5
LDL 70 mg/dl, CRP 2 mg/L
LDL 70 mg/dl, CRP <2 mg/LLDL <70 mg/dl, CRP 2 mg/L
LDL <70 mg/dl, CRP <2 mg/L
LDL <70 mg/dl, CRP <1 mg/L
0.00
0.02
0.04
0.06
0.08
0.10
Ridker PM et al. N Engl J Med 2005;352:20-28. Copyright 2005 Massachusetts Medical Society. All rights reserved.
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4
JUPITER: Primary endpoint
Years
Cumulative incidence
0.08
0.06
0.04
0.02
0.000 1 2 3 4
Placebo
Rosuvastatin
Ridker PM et al. N Engl J Med. 2008;359:2195-207.
HR 0.56 (0.46-0.69)P < 0.00001
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5
JUPITER: MI, stroke, or CV death
Ridker PM et al. N Engl J Med. 2008;359:2195-207.
Placebo
Rosuvastatin
Years
Cumulative incidence
0.08
0.06
0.04
0.02
0.000 1 2 3 4
HR 0.53 (0.40-0.69)P < 0.00001
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6
N Rate
Placebo 7832 1.11LDL-C achieved 70 mg/dL 2110 0.91 P < 0.0001LDL-C achieved <70 mg/dL 5606 0.51
Placebo 7832 1.11LDL-C reduction <50% 4181 0.74 P < 0.0001LDL-C reduction 50% 3535 0.47
Placebo 7832 1.11hsCRP achieved 2 mg/L 4305 0.77 P < 0.0001hsCRP achieved <2 mg/L 3411 0.42
Placebo 7832 1.11hsCRP reduction <50% 4143 0.70 P < 0.0001hsCRP reduction 50% 3573 0.51
JUPITER: Clinical events according to magnitude of reduction in LDL-C or hsCRP
Ridker PM et al. Lancet. 2009;373:1175-82.
MI, stroke, admission for UA, arterial revascularization, or CV death
Rosuvastatinbetter
Rosuvastatinworse
0.25 0.5 1.0 2.0 4.0
Hazard ratio (95% CI)
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7JUPITER: Importance of achieving dual LDL-C and hsCRP reductionCumulative incidence of MI, stroke, admission for UA, arterial revascularization, or CV death
1 2 3 4Follow-up (years)
Cumulative incidence
0.08
0.06
0.04
0.02
0
Rosuvastatin (LDL-C 70 mg/dL or hsCRP 2 mg/L)
Placebo Rosuvastatin (LDL-C <70 mg/dL and hsCRP <2 mg/L)
Ridker PM et al. Lancet. 2009;373:1175-82.
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8Summary of Studies of the Effect of C-Reactive Protein (CRP) Genotype Combination and Apolipoprotein E
Genotype on the Risk of Ischemic Heart Disease (IHD)
Zacho J et al. N Engl J Med 2008;359:1897-1908
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9Summary of Studies of the Effect of C-Reactive Protein (CRP) Genotype Combination on the Risk of Ischemic
Cerebrovascular Disease (ICVD)
Zacho J et al. N Engl J Med 2008;359:1897-1908
Effect of Insulin on NEEffect of Insulin on NE--Induced Induced VenoconstrictionVenoconstriction
A B
DC
Grover A, et al. Hypertension. 1995;25:779-784.
A. Normal vein before cuff inflation
C. Constriction induced by norepinephrine
B. The same vein after inflation
D. Attenuation of the norepin- ephrine effect by insulin
Effect of Insulin on NEEffect of Insulin on NE--Induced Induced Venoconstriction: ControlsVenoconstriction: Controls
Diameter of Vein (%)
Relative to Basal Inflated
Grover A, et al. Hypertension. 1995;25:779-784.
NE (ng/min) NE (100 ng/min) +Insulin (U/min)
Basal
P<0.05
Inflated 12.5 25 50 100 8 16 24 320
20
40
60
80
100
120 100%97%
74%
67%60%
77%80%
87% 97%
Effect of Methylene Blue (MB)Effect of Methylene Blue (MB)
Grover, et al. Hypertension. 1995;25:779-784.
Diameter of Vein (mm)
NE (100 ng/min) NE (100 ng/min) +IN (32 U/min) +MB (0.125 g/min)
NE (100 ng/min) +Insulin (32 U/min)
0.0
0.5
1.0
1.5
2.0
2.5
3.0
Mean ±
SEM
Inability of Insulin to Inhibit Platelet Inability of Insulin to Inhibit Platelet Aggregation in DiabeticsAggregation in Diabetics
Dandona P. Presented at :American Diabetes Association 57th Scientific Session; June, 1997. Boston, Mass.
400 nM U46619 Agonist
CD50 of Insulin(µU/mL)
Control IDDM NIDDM0
10
20
30
40
50
60
70
P<0.001
Factor VIII Ulex Europaeus
CD31 EN4
-Actin Negative Control
Aljada A, Dandona P. Metabolism. 2000;49:147-150.
A) A representative standard curve is shown (r2 = 0.99). B) Representative output generated from an experiment where NO production was measured in response to
increasing concentrations of insulin. Current generated by the NO electrode is recorded as a function of time. The concentrations of insulin shown represent cumulative concentrations.
Direct Measurement of NO From HUVECDirect Measurement of NO From HUVEC
B
Basal
50 pA200 sec
1 50 100 500 1000 5000 10,000 50,000Insulin Dose(nM)
Nitric Oxide(nM)
A 200150100500
0 50 100 150 200 250 300Current (pA)
Zeng G, Quon MJ. J Clin Invest. 1996;98:894-898.
A) Western blot showing the induction of eNOS in human aortic endothelial cells by insulin. The induction is observed at 25 U/mL.
B) No i-NOS bands were observed in HAEC induced with different concentrations of insulin even at very high X-ray film exposure.
A)
Insulin
µU/m
L
µU/m
L
µU/m
L
µU/m
L
0 25 100
1000EC N
OS
140 KD-
B) Insulin
µU/m
L
µU/m
L
µU/m
L
µU/m
L
0 25 100
1000
130 kDa-
LPS-
Stim
ulated
Mono
cytes
Induction of eNOS in Induction of eNOS in Endothelial Cells by InsulinEndothelial Cells by Insulin
Adapted from: Aljada A, Dandona P. Metabolism. 2000;49:147-150.
% C
hang
e Fro
m Ba
selin
e eNO
S Pr
otein
Leve
ls0
50
100
150
200
250
300
350
Insulin (U/ml, Mean
SD)0 25 100 1000
*
*
*
*P<0.05
NFNF--B ActivationB Activation
IB Kinases
Cytoplasm
Nucleus
Inflammatory Gene
mRNA
Degradation
NF-B
Cell Membrane
Activation Signals
Inflammatory Proteins
Barnes and Karin. N Engl J Med. 1997;336:1066-1071.
Barnes and Karin. N Engl J Med. 1997;336:1066-1071.
Effects of Glucocorticoids Effects of Glucocorticoids on NFon NF--B ActivationB Activation
Cytokine
Cytoplasm
Coding Sequence
Inflammatory Gene
mRNA
Nucleus
NF-B
Cell Membrane Glucocorticoid Inflammatory
ProteinsCytokine Receptor
Glucocorticoid Receptor
Glucocorticoid Response ElementIB
Gene
Protein
Plasma Insulin and Glucose Concentrations Following Insulin or Plasma Insulin and Glucose Concentrations Following Insulin or Glucose or Saline Infusions*Glucose or Saline Infusions*
*Insulin infusion in 10 obese human subjects.
Adapted from: Dandona P, et al. J Clin Endocrinol Metab. 2001;86:3257-3265.
Time (hours)0 2 4 6
Insuli
n (μU
/mL)
0
5
10
15
20
25
30
35
40
*P<0.05*
*
Insulin InfusionDextrose InfusionSaline Infusion
Time (hours)0 2 4 6
Plas
ma G
lucos
e (mg
/dL)
0102030405060708090
100110
Insulin InfusionDextrose InfusionSaline Infusion
ROS Generation by MNC in Obese Subjects Following ROS Generation by MNC in Obese Subjects Following Insulin or Dextrose or Saline InfusionsInsulin or Dextrose or Saline Infusions
Adapted from: Dandona P, et al. J Clin Endocrinol Metab. 2001;86:3257-3265.
%RO
S Ge
nera
tion
By M
NC
Time (hours)0 2 4 6
0102030405060708090
100110120130140150160
*P<0.05
*
*
*
Insulin InfusionDextrose InfusionSaline Infusion
Relative Expression of p47Relative Expression of p47phoxphox
Subunit Protein LevelsSubunit Protein Levels
Time (hours): 0 2 4 6
Time (hours): 0 2 4 6
p47phox
p47phox
Mean
SE.
Adapted from: Dandona P, et al. J Clin Endocrinol Metab. 2001;86:3257-3265.
Relative Expression of p47phox
Subunit in MNC Following Insulin Infusion (Upper Gel) or Dextrose
Infusion (Lower Gel)
Densitometry
% C
hang
e In p
47ph
oxSu
bunit
Time (hours)0 2 4 6
0
20
40
60
80
100
120
140
160
180
200
*P<0.05
**
Insulin InfusionDextrose Infusion
Relative NFRelative NF--B Binding to DoubleB Binding to Double--Stranded OligonucleotideStranded Oligonucleotide-- Containing NFContaining NF--B DNA Binding Site Following Insulin or Dextrose B DNA Binding Site Following Insulin or Dextrose
InfusionInfusionGel Shift Assay
Insulin Infusion
Dextrose Infusion
Adapted from Dandona P, et al. J Clin Endocrinol Metab. 2001;86:3257-3265.
% C
hang
e In I
ntran
uclea
r NFk
B
Time (hours)0 2 4 6
0
20
40
60
80
100
120
140Insulin InfusionDextrose Infusion
*P<0.05
*
*
Plasma sICAMPlasma sICAM--1 Concentrations Following Insulin or 1 Concentrations Following Insulin or Dextrose or Saline InfusionsDextrose or Saline Infusions
Adapted from Dandona P, et al. J Clin Endocrinol Metab. 2001;86:3257-3265.
% Change in sICAM-1
(ng/mL)
Time (hours)0 2 4 6
50
60
70
80
90
100
110
120
130
140
**
Insulin InfusionDextrose InfusionSaline Infusion
*P<0.05
Plasma MCPPlasma MCP--1 Concentrations Following Insulin or 1 Concentrations Following Insulin or Dextrose or Saline InfusionsDextrose or Saline Infusions
Adapted from Dandona, et al. J Clin Endocrinol Metab. 2001;86:3257-3265.Time (hours)
0 2 4 650
60
70
80
90
100
110
120
130
140
150
160
% C
hang
e In M
CP-1
*P<0.05
*
*
Insulin InfusionDextrose InfusionSaline Infusion
Plasma MMPPlasma MMP--2 and MMP2 and MMP--9 Concentrations 9 Concentrations Following Insulin or Dextrose InfusionsFollowing Insulin or Dextrose Infusions
Dandona P, Aljada A, et al. Diabetes Care. 2003;26:3310-3314.
Time (hours)0 2 4 6
% C
hang
e in M
MP-2
0
20
40
60
80
100
120
140
160
**
Insulin InfusionDextrose Infusion
*P<0.05
% C
hang
e in M
MP-9
0
20
40
60
80
100
120
140
160
*P<0.05
Time (hours)0 2 4 6
* *
Insulin InfusionDextrose Infusion
Plasma TF and PAIPlasma TF and PAI--1 Concentrations Following Insulin or 1 Concentrations Following Insulin or Dextrose InfusionsDextrose Infusions
Time (hours)0 2 4 6
% C
hang
e in P
lasma
TF
0
20
40
60
80
100
120
140
160
180
200
**
Insulin InfusionDextrose Infusion
*P<0.05
% C
hang
e in P
lasma
PAI
-1
0
20
40
60
80
100
120
140
160
180
200
*P<0.05
Time (hours)0 2 4 6
* **
Insulin InfusionDextrose Infusion
Plasma VEGF Concentrations Following Insulin or Plasma VEGF Concentrations Following Insulin or Dextrose InfusionsDextrose Infusions
% Changein VEGF
0
20
40
60
80
100
120
140
160
Time (hours)0 2 4 6
*P<0.05
* *
Insulin InfusionDextrose Infusion
Dandona P, Aljada A, et al. Diabetes Care. 2003;26:3310-3314.
Toll-Like Receptors (TLRs) Family
Effect of Insulin Infusion in T2DM onTLR4 Expression
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Hours0 2 4 6
% C
hang
e in
TLR
4 m
RN
A E
xpre
ssio
n
40
60
80
100
120
140 InsulinDextrose
*#
*#
Hours0 2 4 6
% C
hang
e in
TLR
2 m
RN
A E
xpre
ssio
n
60
80
100
120
140
160
InsulinDextrose
*# *
Effect of Insulin Infusion in T2DM onTLR2 Expression
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
GIKGIK-- MIMI-- GlucoseGlucose
Time (hours)0h 2h 4h 6h 24h 48h
Gluc
ose (
mg/dl
)
40
60
80
100
120
140
160
180
200
220
Control Insulin
GIKGIK-- MIMI-- InsulinInsulin
*p< 0.05, TWANOVA, † p<0.05, paired t-test
Insuli
n (µU
/ml)
Time (hours)0h 2h 4h 6h 24h 48h
20
40
60
80
100
*††
†
††
Control Insulin
GIKGIK-- MIMI-- Plasma CRPPlasma CRP
*p< 0.01, TWANOVA, † p< 0.05, t-testTime (hours)
0h 2h 4h 6h 24h 48h
Incre
ase i
n CRP
(ng/m
l) fro
m ba
selin
e
0
-1000
1000
2000
3000
4000
5000
6000
7000*†
†Control Baseline: 2700ng/ml Insulin Baseline: 3800ng/ml
*p< 0.01, TWANOVA, † p< 0.05, t-test
GIK GIK –– MI Serum Amyloid AMI Serum Amyloid AInc
reas
e in S
AA (n
g/ml) f
rom
base
line
0
10x103
20x103
30x103
Time (hours)0h 2h 4h 6h 24h 48h
*†
†Control Baseline : 12 x 103
Insulin Baseline: 19 x 103
*p< 0.05, TWANOVA
GIKGIK-- MIMI-- Plasma PAIPlasma PAI--11Inc
reas
e in P
AI-1
(ng/m
l) Fro
m Ba
selin
e
Time (hours)0h 2h 4h 6h 24h 48h
-10
0
10
20
*
Control Baseline: 47ng/ml Insulin Baseline: 54 ng/ml
*p< 0.05, TWANOVA, † p<0.05, t-test, ‡ p< 0.05, paired t - test
GIKGIK-- MIMI-- p47p47phoxphox SubunitSubunit
0h 2h 4h 6h 24h 48h0
200
400
600
% In
creas
e in P
47 ph
ox P
rotei
n in M
NC
Time (hours)
*
†
‡
Control Insulin
*p<0.01, Log CK, TWANOVA
GIKGIK-- MIMI-- Inferior Wall CKInferior Wall CKCK
(U/L)
Time (hours)0h 2h 4h 6h 8h 16h 24h 48h
0
500
1000
1500
2000
*
Control Insulin
*p<0.01, Log CKMB, TWANOVA
GIKGIK-- MIMI-- Inferior Wall CKMBInferior Wall CKMBCK
MB (U
/L)
Time (hours)0h 2h 4h 6h 8h 16h 24h 48h
-100
0
100
200
300
400
500
*
Control Insulin
GIKGIK-- MIMI-- Myoglobin
Time (hours)
0h 2h 4h 6h 24h 48h
Cha
nge
in M
yogl
obin
(ng/
ml)
from
bas
elin
e
-300-250-200-150-100-50
050
100150200250300350400
insulin control
*
Time (hours)
0h 2h 4h 6h 24h 48h
Free
Fat
ty A
cids
(mic
rom
ol/l)
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
control Insulin
*
GIKGIK-- MIMI-- FFA
GIK = glucose-insulin-potassium; CABG = coronary artery bypass graft.Visser L et al. Br J Anaesth. 2004;109:849-854.
Effects of Insulin Infusion Clamp on Effects of Insulin Infusion Clamp on Inflammatory Responses During CABGInflammatory Responses During CABG
*†
†
ControlGIK
250
200
150
100
50
0
CR
P (m
g/L)
Baseli
neRep
erfusio
n2-h
Rep
erfus
ionIC
U 6-8 P
MPOD1 6
-8 AM
POD1 6-8
PMPOD2 6
-8 AM
POD2 6-8
PM
Time dependence of CRP production. Data are presented as mean (SEM). *P<0.05 between groups; 2-way ANOVA for repeated measurements with Bonferroni correction; †P<0.05 between groups at separate time points. Note that at some measurement points the error bars are too small to be visible on the scale used.
Admission Glucose Dependent Mortality in the CREATE-ECLA
6.6
8.5
14.0
<126126to
144
>144
Glucose LevelsGlucose LevelsCONTROL(mg/dl)
GIK(mg/dl)
Baseline 162 162
6 hrs 145 187
24hrs 135 160
Estimated 30 day mortality versus Baseline Blood Glucose Level iEstimated 30 day mortality versus Baseline Blood Glucose Level in n
CREATECREATE--ECLAECLA
Estimated 30 Day Mortality versus Baseline Blood Glucose Level
4
5
6
7
8
9
10
11
12
13
14
15
5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11
Blood Glucose (mmol/L)
Mor
taili
ty (%
)
% mortality = 100 * {1 - 1.03*exp(-0.0167*BG) }
Based on JAMA 1/26/05-Vol 293, No 4, pg 443
30 day mortaility = 13.4% at BG = 187mg/dl
30 day mortaility = 10.8% at BG = 145mg/dl
Chaudhuri A et al: Targeting Glucose in AMI: Has GIK missed the target? (Diabetes Care, 2007)
Hyperglycemia related mortality inHyperglycemia related mortality in CREATE ECLA based CREATE ECLA based on relationship of admission glucose to mortality in on relationship of admission glucose to mortality in
controls.controls.
Control GIK
Time BG*(mg/dl)
% Mortality BG*(mg/dl)
% Mortality
0 162 11.4 162 11.4
6h 145 10.2 187 13.4
24h 135 9.1 160 10.8
Chaudhuri A et al: Targeting Glucose in AMI: Has GIK missed the target? (in press), 2007
Observed vs Estimated Mortality based on blood Observed vs Estimated Mortality based on blood glucose during the 24 hrs in CREATE ECLAglucose during the 24 hrs in CREATE ECLA
02468
101214
Control GIK
ObservedEstimated
9.7% 9.9% 10.0%
12.2%
Chaudhuri A et al: Targeting Glucose in AMI: Has GIK missed the target? (Diabetes Care, 2007
INTENSIVE Study: Intensive Insulin Therapy and Size of INTENSIVE Study: Intensive Insulin Therapy and Size of Infarct as a Visual Endpoint by Cardiac MRIInfarct as a Visual Endpoint by Cardiac MRI
TIMI = thrombolysis in MI; TMPG = TIMI myocardial perfusion grade; STR = ST segment recovery.
RevascularizationI
Inflammatory Markers
Intensive Insulin Therapy (n = 294) BG goal 90-130
Hour 0 Day 6096 hours
Men or Women
35-80 y/oPrimary AWMI
Glucose 140 mg/dL<6 hours of symptoms
TIMI flow, TMPG, STR
Primary Endpoint: Final Myocardial Infarct Size at Day 60 (DE- CMR)
Randomization
Standard Care (n = 294) BG goal <180
MACE : Major Adverse Coronary Events
DE-CMR = Delayed Enhancement Cardiovascular Magnetic Resonance
Time (Hours)-2 0 2 4 6 8 10 12 14 16 18 20 22 24 26
% C
hang
e in
Pla
sma
NO
2/NO
3
60
80
100
120
140
160
180
200
LPS+ SalineLPS+Insulin
Ins/Sal
LPS
*#
*#
*#
*#
Insulin Infusion Suppresses LPS-
Induced Increase in NO2
/NO3
Concentrations
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Insulin Infusion Suppresses LPS-
Induced Increase in ROS Generation by PMN
Tim e (H ours)-1 0 1 2 4 6 24
% C
hang
e in
RO
S G
ener
atio
n (P
MN
)
0255075
100125150175200225250275300325350375400
LPS (2ng/Kg)LPS (2ng/Kg)+ Ins
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Insulin Infusion Suppresses LPS-
Induced Increase in Plasma TBARS
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVAT im e (H ours)
0 1 2 4 6 24
% C
hang
e in
Pla
sma
TB
AR
S
50
100
150
200
250
300
350
400L P SL P S +In s
Insulin Infusion Suppresses LPS-
Induced
Increase in MIF Concentrations
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Time (Hours)0 1 2 4 6 24
% C
hang
e in
MIF
Con
cent
ratio
ns
60
80
100
120
140
160
180
200
220
240
260 EndotoxinEndotoxin+Insulin
Insulin Infusion Suppresses LPS-
Induced
Increase in Myoglobin
Concentrations
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVATime (Hours)
0 1 2 4 6 24
% C
hang
e in
Myo
glob
in C
once
ntra
tions
40
60
80
100
120
140
160
180
200
220
EndotoxinEndotoxin+Insulin
Insulin Infusion Suppresses LPS-
Induced
Increase in FFA Concentrations
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVATim e (H ours)
0 1 2 4 6 24
Pla
sma
FFA
(%
Ch
ange
)
0
20
40
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340EndotoxinEndotoxin+Insulin
Effect of Low Dose Insulin Infusion in T2DM onAmyloid-
Precursor mRNA Expression in MNC
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVAHours
0 2 4 6
% C
hang
e in
APP
m
RN
A E
xpre
ssio
n
50
60
70
80
90
100
110
120
130
140InsulinDextroseSaline
*#
Effect of Low Dose Insulin Infusion in T2DM onIL4 mRNA Expression in MNC
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Infusion time (hours)0 2 4 6
% C
hang
e in
IL-4
mR
NA
Exp
ress
ion
40
60
80
100
120
140 InsulinDextroseSaline
*#
*#
Effect of Low Dose Insulin Infusion in T2DM onADAM-33 Expression in MNC
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVAHours
0 2 4 6
% C
hang
e in
AD
AM
33 m
RN
A E
xpre
ssio
n
50
60
70
80
90
100
110
120
130
140InsulinDextroseSaline
*# *
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Effect of Low Dose Insulin Infusion in T2DM onCCR2 mRNA Expression in MNC
Hours0 2 4 6
% C
hang
e in
CC
R-2
mR
NA
Exp
ress
ion
40
50
60
70
80
90
100
110
120 InsulinDextrose
*# *#
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Infusion time (hours)0 2 4 6
% C
hang
e in
CC
R5
mR
NA
Exp
ress
ion
40
60
80
100
120
140 InsulinDextrose
*# *#
Effect of Low Dose Insulin Infusion in T2DM onCCR5 mRNA Expression in MNC
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Effect of Low Dose Insulin Infusion in T2DM onPlasma MCP-1 Concentrations
Hours0 2 4 6
% C
hang
e in
pla
sma
MC
P-1
con
cent
ratio
ns
60
70
80
90
100
110
120
130InsulinDextroseSaline
*# *
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Effect of Low Dose Insulin Infusion in T2DM onPlasma Eotaxin Concentrations
Hours0 2 4 6
Eota
xin
Con
cent
ratio
ns (%
Cha
nge)
70
75
80
85
90
95
100
105
110
115
120 InsulinDextroseSaline
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Effect of Low Dose Insulin Infusion in T2DM onPlasma RANTES Concentrations
Hours0 2 4 6
Plas
ma
RA
NTE
S C
once
ntra
tions
(ng/
ml)
20
25
30
35
40
45 InsulinDextroseSaline
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA
Effect of Low Dose Insulin Infusion in T2DM onPlasma MMP-9 Concentrations
Hours0 2 4 6
% C
hang
e in
MM
P-9
Plas
ma
conc
entr
atio
ns
60
70
80
90
100
110
120InsulinDextroseSaline
*#
*
Hours0 2 4 6
% C
hang
e in
Pla
sma
NO
2/NO
3
40
50
60
70
80
90
100
110
120
130
140InsulinsalineDextrose
*#
Effect of Low Dose Insulin Infusion in T2DM onPlasma NO2 /NO3 Concentrations
* P < 0.05 by One Way RM ANOVA # P < 0.05 by Two Way RM ANOVA