RESUMO
At this centenary of the British Journal of Anaesthesia (BJA) in 2023, six of its 12 editors/editors-in-chief detail developments over the decades that have led to the BJA becoming a high-impact international scientific journal. As a charity, the BJA supports academic research and training in anaesthesia, critical care, and pain medicine including funding of research grants and postgraduate education. Building on this foundation, the BJA continues to innovate as it aims to become fully electronic, expand into open access publishing, and increase the diversity of its editorial board.
Assuntos
Anestesia , Anestesiologia , Humanos , Cuidados CríticosRESUMO
Microvascular inflammation occurs during sepsis and the endogenous opioid-like peptide nociceptin/orphanin FQ (N/OFQ) is known to regulate inflammation. This study aimed to determine the inflammatory role of N/OFQ and its receptor NOP (ORL1) within the microcirculation, along with anti-inflammatory effects of the NOP antagonist UFP-101 (University of Ferrara Peptide-101) in an animal model of sepsis (endotoxemia). Male Wistar rats (220 to 300 g) were administered lipopolysaccharide (LPS) for 24 h (-24 h, 1 mg kg(-1); -2 h, 1 mg kg(-1) i.v., tail vein). They were then either anesthetised for observation of the mesenteric microcirculation using fluorescent in vivo microscopy, or isolated arterioles (~200 µm) were studied in vitro with pressure myography. 200 nM kg(-1) fluorescently labelled N/OFQ (FITC-N/OFQ, i.a., mesenteric artery) bound to specific sites on the microvascular endothelium in vivo, indicating sparse distribution of NOP receptors. In vitro, arterioles (~200 µm) dilated to intraluminal N/OFQ (10(-5)M) (32.6 + 8.4%) and this response was exaggerated with LPS (62.0 +7.9%, p=0.031). In vivo, LPS induced macromolecular leak of FITC-BSA (0.02 g kg(-1) i.v.) (LPS: 95.3 (86.7 to 97.9)%, p=0.043) from post-capillary venules (<40 µm) and increased leukocyte rolling as endotoxemia progressed (p=0.027), both being reduced by 150 nmol kg(-1) UFP-101 (i.v., jugular vein). Firstly, the rat mesenteric microcirculation expresses NOP receptors and secondly, NOP function (ability to induce dilation) is enhanced with LPS. UFP-101 also reduced microvascular inflammation to endotoxemia in vivo. Hence inhibition of the microvascular N/OFQ-NOP pathway may have therapeutic potential during sepsis and warrants further investigation.
Assuntos
Inflamação/patologia , Antagonistas de Entorpecentes , Peptídeos Opioides/farmacologia , Animais , Células CHO , Sistema Cardiovascular/efeitos dos fármacos , Sistema Cardiovascular/patologia , Sistema Cardiovascular/fisiopatologia , Cricetinae , Cricetulus , Fluoresceína-5-Isotiocianato/metabolismo , Inflamação/metabolismo , Migração e Rolagem de Leucócitos/efeitos dos fármacos , Masculino , Microcirculação/efeitos dos fármacos , Microvasos/efeitos dos fármacos , Microvasos/patologia , Peptídeos Opioides/metabolismo , Ratos , Ratos Wistar , Receptores Opioides/metabolismo , Proteínas Recombinantes/metabolismo , Receptor de Nociceptina , NociceptinaRESUMO
The spleen has an important role in blood volume regulation and increased resistance of post-capillary hilar veins (in mesentery adjoining the spleen) can regulate this. This study investigated whether venular constriction to lipopolysaccharide (LPS) involved endothelin-1 (ET-1). Pressure myography was used to study isolated extra-splenic (hilar) vessels from male Wistar rats (n = 111). Arteries and veins were treated with LPS (50 microg ml(-1)) for 4 h. Extra-splenic veins constricted to LPS (p < 0.05), but there was no effect on arteries. Denudation did not abolish venular constriction to LPS, indicating an endothelial independent mechanism. However, the dual ET-1 receptor antagonist bosentan (10(-5) M) and specific ET(A) and ET(B) antagonists ABT-627 (atrasentan, 6.3 x 10(-6) M) and A-192621(1.45 x 10(-6) M) completely abolished constriction of LPS-treated veins. ET-1 alone also constricted the extra-splenic arteries and veins (p < 0.05), with a greater response observed in veins (p < 0.05). ELISA also confirmed that serum and spleen levels of ET-1 increased in response to LPS (p < 0.05). That LPS-induced constriction of extra-splenic veins is mediated by ET-1. Greater constriction of post- versus pre-capillary extra-splenic vessels to LPS would result in increased intra-splenic fluid extravasation and hypovolaemia in vivo.
Assuntos
Endotelina-1/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , Receptor de Endotelina A/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacos , Animais , Atrasentana , Bosentana , Endotelina-1/metabolismo , Ensaio de Imunoadsorção Enzimática , Hipovolemia/etiologia , Masculino , Mesentério/metabolismo , Miografia , Pirrolidinas/farmacologia , Ratos , Ratos Wistar , Receptor de Endotelina A/metabolismo , Baço/efeitos dos fármacos , Baço/metabolismo , Sulfonamidas/farmacologiaRESUMO
BACKGROUND: The spleen has an important physiological role in maintaining blood volume; this study aimed to determine whether during pathophysiological circumstances, namely endotoxemia, the extrasplenic pathway is dysfunctional. We hypothesize that increased 'leakiness' of lymphatics in response to lipopolysaccharide (LPS) provides a route for loss of protein-rich fluid into third spaces and prevents the spleen from maintaining blood volume homeostasis. METHODS AND RESULTS: Male Wistar rats (200-280 g, n = 24) were anesthetized with thiopental (40-90 mg x kg(-1) x hr(-1), i.v.) to study the extrasplenic (vessels in mesentery adjoining the spleen) and ileal mesenteric microcirculation using fluorescently labeled albumin (66 KDa FITC-BSA, 0.02 g.100 g(-1), i.v.) with intravital microscopy. LPS (150 microg x kg(-1) x hr(-1) i.v.) induced constriction of rat extrasplenic venules (-14 +/- 2.4% from 40.4 +/- 7.8 microm, p < 0.05) and no change in arteriolar diameter (-4.6 +/- 4.7% from 32.6 +/- 4.3 microm). As the spleen is freely permeable to protein, a greater increase in venular versus arteriolar extrasplenic resistance increases intrasplenic capillary hydrostatic pressure, leading to fluid efflux into the lymphatics, draining the spleen. In agreement we report here increased extrasplenic venular resistance with LPS and lymphatic dilation to accommodate this fluid (13.5 +/- 6% from 18.5 +/- 4.8 microm, p < 0.05). However, the extrasplenic pathway then appeared to dysfunction, with macromolecular leak from extrasplenic venules (24.6 +/- 6.4%, p < 0.05) and lymphatics (12.1 +/- 3.4%, p < 0.05), indicated by increased interstitial FITC-BSA fluorescence. This was less than from ileal mesenteric venules (324 +/- 32%, p < 0.05). There was a concurrent decrease in mean arterial pressure (T(180): -15.1 +/- 6.9% from MAP of 130.3 +/- 8.8 mmHg at T(0), p < 0.05). CONCLUSION: Lymphatics are generally considered to demonstrate unidirectional and inward uptake of large molecules. However, during endotoxemia, we have demonstrated that extrasplenic lymphatics also allow the leakage of large protein molecules out into interstitial spaces. Fluid losses from extrasplenic lymphatics could therefore contribute to hypovolemia and hypotension associated with sepsis.
Assuntos
Endotoxemia/fisiopatologia , Lipopolissacarídeos/química , Vasos Linfáticos/fisiopatologia , Animais , Pressão Sanguínea , Corantes Fluorescentes/farmacologia , Homeostase , Masculino , Microcirculação , Proteínas/química , Ratos , Ratos Wistar , Sepse/fisiopatologia , Baço/metabolismo , Baço/fisiologia , Fatores de TempoRESUMO
Nitric oxide (NO) induces vascular relaxation via cGMP in vascular smooth muscle (VSM) and is an important mediator of vascular tone during sepsis, as endothelial NO synthase (eNOS) may be upregulated during the early stages. Atrial natriuretic peptide (ANP) also stimulates cGMP via eNOS hence, this study aimed to investigate the role of NO in time-dependent altered vascular responses to ANP during the first 4h of exposure to bacterial lipopolysaccharide (LPS). We used male rat saphenous arteries [internal relaxed diameter 63-152 microm, n=48], mounted on a wire myograph and pre-constricted with phenylephrine. At 2h in the presence of LPS, there was increased relaxation to ANP in arteries exposed to LPS [16.3+/-2.4%, P<0.05]. However the response to ANP was not altered by the NOS inhibitor Nomega-nitro-l-arginine methyl ester (L-NAME, 10(-4)M) and following denudation (vessels without endothelium). At 4h there was no longer increased relaxation to ANP in the presence of LPS. Moreover the vasodilator response to ANP was significantly reduced following L-NAME or denudation [4.4+/-1.0% and 4.3+/-1.1% respectively, P<0.05]. However, the non-specific endothelin-1 (ET-1) receptor antagonist Bosentan [10(-5)M] increased dilatation in LPS exposed arteries at 1 and 2h, reaching significance at 4h [14.0+/-3.4%, P<0.05]. In summary, an endothelial and NO dependent mechanism is responsible for increased relaxation to ANP following 2h exposure to LPS. However after 4h an endothelial and NO independent process involving ET-1 is responsible for decreased relaxation to ANP. The enhanced response to ANP may exacerbate early systemic vasodilatation during early sepsis.
Assuntos
Fator Natriurético Atrial/farmacologia , Endotelina-1/metabolismo , Lipopolissacarídeos/farmacologia , Óxido Nítrico/metabolismo , Vasodilatação/efeitos dos fármacos , Animais , Bosentana , Antagonistas do Receptor de Endotelina A , Humanos , Técnicas In Vitro , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Ratos , Ratos Wistar , Veia Safena/efeitos dos fármacos , Veia Safena/metabolismo , Veia Safena/fisiologia , Sulfonamidas/farmacologia , Fatores de Tempo , Vênulas/efeitos dos fármacos , Vênulas/metabolismo , Vênulas/fisiologiaRESUMO
Nitric oxide (NO) and endothelin 1 (ET-1) increase significantly during the first 4 h of Escherichia coli lipopolysaccharide (LPS) exposure. The aim of this study was to investigate the role of these mediators in the reduced response to phenylephrine treatment. We used male rat saphenous arteries (internal relaxed diameter, 63-152 microm; n = 48), mounted on a wire myograph and subsequently treated with LPS. At 1 h, LPS (dose, 50 microg mL(-1)) significantly (P < 0.05) inhibited constriction to phenylephrine (concentration, 10(-1)M to 10(-6)M) (LPS concentration required for half maximal response [EC50], 10.82 +/- 1.08microM; Control EC50, 5.07 +/- 0.34microM). However, by removing the endothelium (denuded) or adding Nomega-nitro-L-arginine methyl ester (L-NAME; concentration, 10(-4) microM), the response to phenylephrine treatment was significantly improved compared with LPS only-treated arteries (LPS + denuded EC50, 7.04 +/- 1.12microM; LPS + L-NAME EC50, 2.64 +/- 0.63microM). On the other hand, denudation did not restore constriction to phenylephrine at 2 and 4 h. However, L-NAME and the nonspecific ET-1 receptor antagonist bosentan (concentration, 10(-5)M) improved constriction to phenylephrine in LPS-treated arteries (P < 0.05) at 4 h (LPS EC50, 998.50 +/- 447.10microM; LPS + L-NAME EC50, 65.23 +/- 25.61microM; LPS + bosentan EC50, 63.65 +/- 25.33microM). We conclude that endothelium-dependent mechanisms have an early role in the reduced responsiveness of vascular smooth muscle to vasoconstrictors during simulated septic conditions. Shortly after exposure to LPS (duration, 1 h), endothelium-derived NO seemed to have a role in reduced arterial constriction to phenylephrine, but later (4 h) ET-1 and endothelium-independent increase in NO seemed to contribute further to the loss of response.
Assuntos
Endotelina-1/fisiologia , Lipopolissacarídeos/toxicidade , Óxido Nítrico/fisiologia , Fenilefrina/farmacologia , Vasoconstrição/efeitos dos fármacos , Vasoconstrição/fisiologia , Animais , Inibidores Enzimáticos/farmacologia , Técnicas In Vitro , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Ratos , Ratos Wistar , Vasoconstritores/farmacologiaRESUMO
Sepsis causes changes in vascular resistance and hypovolemia. Previous studies have demonstrated that the spleen regulates blood volume via atrial natiuretic peptide (ANP). We hypothesized that LPS alters extrasplenic responses to ANP via endothelial-dependent mechanisms and studied the role of NO and endothelin 1 (ET-1). Isolated extrasplenic arteries and veins (vessels in mesentery adjoining spleen) were obtained from male Wistar rats weighing 200 to 280 g (n = 102) and mounted on a pressure myograph to determine intraluminal diameter for 4 h. Isolated vessels constricted in response to the half-maximum response of ANP (veins, 30% +/- 1.7%; arteries, 34.5 +/- 1.7%; P < 0.05), and this was abolished by the NO donor S-nitroso-N-acetylpenicillamine (SNAP 75 microM). Arteries and veins incubated with LPS (50 microg mL(-1) for 4 h) were unresponsive to ANP, and constriction was not restored by the NOS inhibitor N omega-nitro-L-arginine methyl ester (L-NAME 100 microM). However, venular constriction returned in the presence of the ET-1 antagonist Bosentan, increasing from -1.5 +/- 1.2 (10 min) to -10 +/- 2.5% (4 h) with LPS + Bosentan (3 x 10(-6) M) compared with -2.3 +/- 1.2 and 0% with LPS alone. In conclusion, LPS abolished endothelial-dependent extrasplenic venular constriction to ANP partially due to increased ET-1, whereas NO seemed to modulate vascular responses to ANP.
Assuntos
Fator Natriurético Atrial/metabolismo , Endotelina-1/metabolismo , Lipopolissacarídeos/toxicidade , Óxido Nítrico/metabolismo , Sepse/metabolismo , Baço/metabolismo , Animais , Anti-Hipertensivos/farmacologia , Artérias/metabolismo , Artérias/fisiopatologia , Volume Sanguíneo/efeitos dos fármacos , Bosentana , Inibidores Enzimáticos/farmacologia , Hipovolemia/induzido quimicamente , Hipovolemia/metabolismo , Hipovolemia/fisiopatologia , Masculino , Mesentério/metabolismo , Mesentério/fisiopatologia , NG-Nitroarginina Metil Éster/farmacologia , Doadores de Óxido Nítrico/farmacologia , Penicilamina/análogos & derivados , Penicilamina/farmacologia , Ratos , Ratos Wistar , Sepse/induzido quimicamente , Sepse/fisiopatologia , Baço/fisiopatologia , Sulfonamidas/farmacologia , Resistência Vascular/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacos , Veias/metabolismo , Veias/fisiopatologiaRESUMO
UNLABELLED: Hemorrhage increases adhesion of leukocytes to the venular endothelium, mediated by increased expression of the Mac-1 integrin complex (CD18/CD11b) present on leukocytes. Anesthetic agents may possess anti-inflammatory properties. Hence, this study determined the effects of i.v. anesthesia on leukocyte adhesion after hemorrhage in relation to expression of CD11b. METHODS: Male Wistar rats were (n = 57) anesthetized i.v. with propofol (Diprivan) and fentanyl, ketamine, or thiopental. During anesthesia, 10% of total blood volume was removed and intravital microscopy used to observe the rat mesentery and measure leukocyte (neutrophils) rolling and adhesion in postcapillary venules (15 - 25 microm). Flow cytometry was also used to determine CD11b expression on neutrophils from blood removed at the end of these experiments (n = 25) or blood incubated with anesthetic agents and activated with platelet activating factor ex vivo (0.1 micromol/L) (n = 24). RESULTS: Hemorrhage increased leukocyte adhesion (stationary count per 150 microm) in rats anesthetized with thiopental (baseline, 3.4 +/- 1.2; hemorrhage, 6.7 +/- 2.0; P < 0.05) but not in those receiving either ketamine (baseline, 3.6 +/- 1.3; hemorrhage, 3.3 +/- 1.3) or propofol/fentanyl (baseline, 6.2 +/- 2.0; hemorrhage, 5.8 +/- 0.8). Neutrophils collected from thiopental-treated rats had elevated CD11b expression with thiopental (mean fluorescence baseline, 67.5 +/- 1.3; hemorrhage, 83.6 +/- 5.3; P < 0.05) but not with propofol/fentanyl (mean fluorescence baseline, 69.1 +/- 1.3; hemorrhage, 65.9 +/- 1.6), and ketamine-treated rats (mean fluorescence baseline, 74.3 +/- 2.1; hemorrhage, 74.8 +/- 1.1). Ketamine also inhibited upregulation of CD11b with platelet activating factor ex vivo. CONCLUSIONS: After hemorrhage, leukocyte adhesion and CD11b expression increased during thiopental anesthesia, but propofol/fentanyl and ketamine protected against hemorrhage-induced leukocyte adhesion. The anti-inflammatory effect of ketamine was mediated by direct inhibition of CD11b expression on leukocytes.
Assuntos
Anestesia Intravenosa , Antígeno CD11b/biossíntese , Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Hemorragia/metabolismo , Leucócitos/metabolismo , Animais , Adesão Celular , Endotélio Vascular/patologia , Inflamação , Masculino , Propofol/farmacologia , Ratos , Ratos Wistar , Tiopental/farmacologia , Fatores de TempoRESUMO
Determining the effects of analgesia on the microcirculation is difficult because the surgery needed to allow in vivo observation often requires anesthesia. In this study, we used the dorsal microcirculatory chamber (DMC) to determine the effects of large (LF) and small (SF) dose IV fentanyl on the microcirculation compared with a conscious control. Male Wistar rats (130 g, n = 5) were implanted with the DMC to enclose a single layer of striated muscle. Animals were allowed 3 wk to recover from surgery and then, over the following 2 wk (1 infusion/wk) using intravital microscopy, the microcirculation was viewed in conscious animals (t = 0-30 min), followed by an induction bolus dose (t = 40-45 min), then a "step-up" maintenance infusion of one of the following, LF (40-90 microg x kg(-1) x h(-1)), SF (10-60 microg x kg(-1) x h(-1)), or saline (5-10 microg x kg(-1) x h(-1)) (t = 45-105 min). Small arterioles (<30 micro m) dilated (23.6% +/- 7.1%) after induction with LF, but constricted (-21.3% +/- 7.1%) with SF (P < 0.05). During maintenance, constriction increased with increasing dose of LF (-21.9% +/- 4.0%) and SF (-16.7% +/- 9.1%) (t = 105 min, P < 0.05). Similar patterns were observed in all arterioles (10-120 microm) and venules (15-250 microm). We conclude that the DMC provides an excellent technique for observing microcirculatory responses to fentanyl, and in rat skeletal muscle in vivo, an i.v. infusion of fentanyl produces significant constriction of arterioles.
Assuntos
Analgésicos Opioides/farmacologia , Fentanila/farmacologia , Músculo Esquelético/irrigação sanguínea , Animais , Arteríolas/anatomia & histologia , Arteríolas/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Relação Dose-Resposta a Droga , Frequência Cardíaca/efeitos dos fármacos , Masculino , Microcirculação/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , Medição da Dor/efeitos dos fármacos , Condicionamento Físico Animal/fisiologia , Ratos , Ratos Wistar , Fluxo Sanguíneo Regional/efeitos dos fármacos , Restrição Física , Vênulas/anatomia & histologia , Vênulas/efeitos dos fármacosRESUMO
Anesthetic agents are known to have differential effects on both the systemic circulation and the microcirculation. The aim of this study was to compare the effects of several intravenous (i.v.) agents on the microcirculatory response to hemorrhage. Male Wistar rats (n = 52) were anesthetized i.v. either with propofol and fentanyl (propofol fentanyl), ketamine, or thiopental. Cardiovascular variables were monitored. The cremaster muscle was observed by using fluorescent intravital microscopy. FITC-BSA was administered (0.25 mL/100 g, i.a.) to determine macromolecular leak, an index of vessel integrity. Animals were further allocated into control (C), 10% hemorrhage (H), or hemorrhage re-infusion (H-R, removal of 10% blood volume and then re-infusion of saline and blood) groups. When systolic arterial pressure (SAP) was maintained after hemorrhage, constriction of A3 and A4 arterioles (5-30 microm) was accompanied by no change in the diameter of A1 (80-130 microm): most frequent with ketamine (A1: -1.7 +/- 1.2; A4: -13.9 +/- 2.7%; H and H-R: n = 9/11). With lower SAP, dilation of the A3 and A4 was accompanied by constriction of the A1: most frequent with propofol/fentanyl (A1: -8.0 +/- 2.5; A4; 35.1 +/- 9.4%; H and H-R: n = 6/11). No increases in macromolecular leak occurred with any anesthetic agent or in H or H-R groups. The response of cremaster muscle microcirculation to hemorrhage differs with different i.v. anesthetic agents. Dilation of small arterioles is the predominant response with propofol/fentanyl and constriction of small arterioles with ketamine.