Background
Methods
Anesthesia and instrumentation
Experimental protocol
Gene expression analysis and immunoblotting
PCR assay | Oligonucleotide sequence (5′-3′) | GenBank no. |
---|---|---|
IL-1β | S: ACCCTgCAgCTggAggAT | NM_214055 |
A: CCTTTggAgTTTCCCAggA | ||
IL-6 | S: CCAATCTgggTTCAATCAggA | NM_214399 |
A: gTggTggCTTTgTCTggATTC | ||
FL: TgTCgAggCTgTgCAgATTAgTACCA(--FL) | ||
CY5: (L670-)gCACTgATCCAgACCCTgAggCAA(--PH) | ||
TNF-α | S: CCCAgAAggAAgAgTTTCCA | NM_214022 |
A: CggCTTTgACATTggCTACA | ||
FL: ggCCCAAggACTCAgATCATCgTC(--FL) | ||
CY5: (L670-)CAAACCTCAgATAAgCCCgTCgC(--PH) | ||
iNOS | S: gATggCACCATCATAggggAC | NM_001143690 |
A: ggCACCCTgggAACTCAA | ||
FL: TGGAACACCCCAAATACGAGTGGTTCC(--FL) | ||
CY5: (L670-)GGAGCTGGAGCTGAAGTGGTACGCCC(--PH) | ||
COX-2 | S: CCCCTTCTgCCTgACgC | NM_214321 |
A: CTCTgCTCTggTCgATTgAgg | ||
FL: TCTATCTTACTggAACATggCATCACCCA(--FL) | ||
CY5: (L670-)TTTgTTgAATCATTTAgCAggCAAATTgCT(--PH) | ||
AREG | S: ATTATGCTGCTGGACTGGAC | NM_214376 |
A: TCGCTACCAGAAGGCATTT | ||
TNC | S: GAGACCTGACTGCTACAGAG | NM_214230 |
A: CACAACGACTTCCTTGAGTG | ||
PPIA | S: CTTTCACAgAATAATTCCAggATT | NM_214353 |
A: ggACAAgATgCCAggACC | ||
FL. ATgCTTCAggATAAAATTCTCATCATCAAA(--FL) | ||
CY5: (L670-)TTCTCTCCATAgATggACTTgCCACCA(--PH) | ||
sENACg | S: ACTTCACCCCCATCTTCCAC | XM_003357614 |
A: TTCAGGCCAAACTCAGCTC |
Hematological parameters
Histopathological and lung water content assessment
Statistical analysis
Results
Pulmonary and systemic inflammatory markers
Physiological data
AP318 | CTRL | |||||||
---|---|---|---|---|---|---|---|---|
Parameter | Baseline | Sepsis /VILI | 3 h | 6 h | Baseline | Sepsis /VILI | 3 h | 6 h |
Ventilation
| ||||||||
V
t
(mL kg
−1
)
| 8.8 ± 1.4 | 25.4 ± 2.0 | 8.7 ± 0.6 | 8.7 ± 0.7 | 8.6 ± 0.9 | 25.8 ± 0.6 | 8.5 ± 0.4 | 8.2 ± 0.6 |
P
endinsp
(cmH
2
O)
| 14 ± 2 | 21 ± 3 | 21 ± 2 | 19 ± 4 | 14 ± 1 | 21 ± 2 | 21 ± 3 | 19 ± 3 |
RR (min
−1
)
| 28 ± 8 | 9 ± 2 | 33 ± 7 | 31 ± 5 | 33 ± 6 | 9 ± 1 | 36 ± 6 | 33 ± 9 |
PEEP (cmH
2
O)
| 6 ± 1 | 1 ± 0 | 5 ± 1 | 5 ± 1 | 6 ± 1 | 1 ± 0 | 5 ± 0 | 6 ± 2 |
FiO
2
| 0.4 | 1.0 | 0.4 | 0.4 | 0.4 | 1.0 | 0.4 | 0.4 |
I:E
| 1:2 | 1:2 | 1:2 | 1:2 | 1:2 | 1:2 | 1:2 | 1:2 |
PaCO
2
(mmHg)
| 43 ± 4 | 35 ± 3 | 44 ± 5 | 42 ± 5 | 45 ± 3 | 36 ± 5 | 43 ± 5 | 42 ± 7 |
pH
| 7.41 ± 0.03 | 7.50 ± 0.08 | 7.33 ± 0.08 | 7.34 ± 0.08 | 7.41 ± 0.04 | 7.47 ± 0.07 | 7.34 ± 0.05 | 7.32 ± 0.08 |
Hemodynamics
| ||||||||
MAP (mmHg)
| 93 ± 13 | 103 ± 11 | 66 ± 13 | 68 ± 8 | 99 ± 13 | 115 ± 12 | 69 ± 12 | 62 ± 12 |
CO (L min
−1
)
| 4.6 ± 0.7 | 4.7 ± 0.7 | 3.2 ± 0.9 | 3.9 ± 1.2 | 4.5 ± 0.5 | 4.6 ± 0.7 | 3.2 ± 0.5 | 3.4 ± 0.7 |
CVP (mmHg)
| 12 ± 3 | 11 ± 2 | 12 ± 3 | 13 ± 2 | 11 ± 3 | 11 ± 3 | 13 ± 3 | 13 ± 3 |
MPAP (mmHg)
| 22 ± 3 | 23 ± 3 | 33 ± 5 | 30 ± 8 | 21 ± 5 | 26 ± 4 | 37 ± 8 | 33 ± 12 |
NA (μg kg
−1
min
−1
) | 0 | 0 | 0.8 ± 1.3 | 2.7 ± 5.0 | 0 | 0 | 0.3 ± 0.5 | 3.1 ± 5.7 |
Pathologic parameters
non-dependent | dependent | |||
---|---|---|---|---|
Parameter | AP318 | CTRL | AP318 | CTRL |
Alveolar edema
| 0 ± 0 | 0.2 ± 0.6 | 0 ± 0 | 0.1 ± 0.2 |
Interstitial edema
| 1.0 ± 0.7 | 1.2 ± 1.1 | 1.0 ± 0.5 | 0.9 ± 0.7 |
Hemorrhage
| 0.3 ± 0.6 | 1.3 ± 1.2* | 0.5 ± 0.8 | 1.2 ± 1.1* |
Inflammatory infiltration
| 3.7 ± 0.9 | 3.5 ± 0.8 | 3.6 ± 1.0 | 3.7 ± 0.8 |
Epithelial destruction
| 0 ± 0 | 0.4 ± 0.9 | 0.1 ± 0.3 | 0.1 ± 0.2 |
Microatelectasis
| 3.6 ± 1.1 | 3.9 ± 1.2 | 3.3 ± 1.4 | 3.4 ± 1.3 |
Overdistension
| 3.9 ± 0.8 | 3.6 ± 1.1 | 3.5 ± 1.0 | 3.5 ± 0.9 |