Supplementary Data
Vasoplegia in sepsis depends on the vascular system, vasopressor, and time-point: a comparative evaluation in vessels from rats subjected to the cecal ligation puncture model
Angélica K. Bernardelli1, 2, Rita de C. V.de A.F. Da Silva1, 2, Thiago Corrêa2 and José Eduardo Da Silva-Santos2*
1Department of Pharmacology, Universidade Federal do Paraná, Curitiba, PR, Brazil;
2Laboratory of Cardiovascular Biology, Department of Pharmacology, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil.
*Corresponding author: Department of Pharmacology, Room 116, Universidade Federal de Santa Catarina, Campus Universitário, Trindade – Florianópolis, SC, 88.040-900, Brazil
Tel: +55 (48) 3721-4847
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Supplementary Methods
Animal survival assessment
The animals underwent ligation surgery and perforation of the cecum and the false-operated animals (sham) underwent the same surgery without ligation or cecum perforation. All animals were evaluated for a period of 96 hours, and survival was recorded at 6, 24, 36, 48 and 96 hours.
Evaluation of systolic pressure
The measurement of systolic blood pressure (SBP) was performed by the tail cuff method. To reduce stress-induced variations in blood pressure, all animals were preconditioned for blood pressure measurements one week before each experiment and the ambient temperature was maintained of 29 ºC. A ten-minute interval for adapting the animal to the procedure was observed, then the sensor (model LE5160R, Panlab, Spain.) was placed in the proximal region of the tail to obtain the SBP values, which was recorded in a digital polygraph (model Power Lab 8/35, AD Instruments, Australia). Three to five measurements were performed in each session, and the average of these measures was used as the systolic blood pressure.
Supplementary Figure Captions
Fig. S1. Survival curve of rats submitted to the cecal ligation and puncture model. The animals were monitored for 96 hours after the procedure (10 animals per group). The results are expressed as percentage (%) surviving.
Fig. S2. Lack of change in vascular reactivity of mesenteric arteries from sham-operated or naïve animals. Mesenteric arteries from naïve or sham-operated animals at 6 h or 18 h after the surgery were stimulated by cumulative concentrations of phenylephrine (A) or norepinephrine (B), or non-cumulative 120 mmol/L KCl (C). The values show the mean ± standard error of the mean of 6–9 preparations per group.
Fig. S3. The mesenteric vascular bed from CLP-subjected rats shows reduced responses to phenylephrine, norepinephrine and angiotensin II, but normal or increased reactivity to vasopressin. The bars show the area under the curve of the effects of non-cumulative doses of phenylephrine (A), norepinephrine (B), angiotensin II (C), and vasopressin (D) obtained in perfused mesenteric arteries from non-septic (sham) and CLP groups at 6 and 18 h after the surgery. The values show the mean ± standard error of the mean of 6 preparations per group. * indicates p < 0.05 compared with the sham group.
Supplementary Figures
Supplementary Table
Table S1. Systolic blood pressure measured by the tail cuff method.
Time (h)Basal values / 3 / 6 / 18
Control / 119.3 ± 4.2 (n=5) / 126.6 ± 4.6 (n=5) / 126.6 ± 4.6 (n=5) / 125.6 ± 5.4 (n=5)
Sham-operated / 117.2 ± 4.4 (n=6) / 104.8 ± 7.8 †‡ (n=6) / 113.6 ± 4.8‡ (n=6) / 115.0 ± 10.0‡ (n=4)
CLP / 118.7 ± 7.8 (n=15) / 81.8 ± 3.3 * (n=8) / 82.2 ± 5.8 * (n=10) / 90.5 ± 4.7 * (n=6)
The values represent the mean ± standard error of the mean. Statistical analyses were performed through two-way ANOVA followed by Tukey's multiple comparison test. * indicates p < 0.05 for CLP compared with the control group; † indicates p < 0.05 for sham-operated compared with the control group; ‡ indicates p < 0.05 for sham-operated compared with the CLP group.