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1.
Medicine (Baltimore) ; 103(17): e37980, 2024 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-38669362

RESUMO

The aim of this observational study was to investigate the effects of catechol-O-methyltransferase (COMT) and ATP-binding cassette transporter B1 (ABCB1) gene polymorphisms on the postoperative analgesic effect of sufentanil in Chinese Han pediatric patients with fractures. A total of 185 pediatric patients who underwent fracture surgery were included. Polymerase chain reaction-restriction fragment length polymorphism was used to detect the polymorphisms of COMT and ABCB1 genes. Sufentanil was used for postoperative analgesia. The pain level of the patients was evaluated using the face, legs, activity, cry, and consolability scale before surgery, during awakening, at 2, 6, 12, and 24 hours after surgery. The postoperative Ramsay sedation score, sufentanil consumption, and incidence of adverse reactions were also recorded. Pediatric patients with different genotypes of ABCB1 and COMT showed no statistically significant differences in general data such as age, gender, weight, height, surgical duration, and American Society of Anesthesiologists classification (P > .05). There were no statistically significant differences in sedation scores after surgery between different genotypes of ABCB1 and COMT (P > .05). Among patients with CC genotype in ABCB1, the pain scores and total consumption of sufentanil at awakening, 2 and 6 hours after surgery were higher compared to TT and CT genotypes (P < .05), while there were no statistically significant differences between TT and CT genotypes (P > .05). Among patients with AA genotype in COMT, the pain scores and total consumption of sufentanil at awakening, 2, 6, 12, and 24 hours after surgery were higher compared to AG and GG genotypes (P < .05), while there were no statistically significant differences between AG and GG genotypes (P > .05). There were no statistically significant differences in adverse reactions between different genotypes of ABCB1 and COMT (P > .05). The polymorphisms of COMT gene rs4680 and ABCB1 gene rs1045642 are associated with the analgesic effect and consumption of sufentanil in pediatric patients after fracture surgery.


Assuntos
Subfamília B de Transportador de Cassetes de Ligação de ATP , Analgésicos Opioides , Catecol O-Metiltransferase , Fraturas Ósseas , Dor Pós-Operatória , Sufentanil , Humanos , Sufentanil/uso terapêutico , Sufentanil/administração & dosagem , Catecol O-Metiltransferase/genética , Dor Pós-Operatória/tratamento farmacológico , Dor Pós-Operatória/genética , Masculino , Feminino , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Criança , Fraturas Ósseas/cirurgia , Fraturas Ósseas/genética , Analgésicos Opioides/uso terapêutico , Analgésicos Opioides/administração & dosagem , Analgésicos Opioides/efeitos adversos , Genótipo , Pré-Escolar , Medição da Dor , Polimorfismo Genético , Adolescente , Polimorfismo de Nucleotídeo Único
2.
Int Immunopharmacol ; 130: 111678, 2024 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-38368773

RESUMO

Aldosterone is a key mineralocorticoid involved in regulating the concentration of blood electrolytes and physiological volume balance. Activation of mineralocorticoid receptor (MR) has been recently reported to participate in adaptive and innate immune responses under inflammation. Here, we evaluated the role of aldosterone and MR in inflammation bowel diseases (IBD). Aldosterone elevated in the colon of DSS-induced colitis mice. Aldosterone addition induced IL17 production and ROS/RNS level in group 3 innate lymphoid cells (ILC3s) and exacerbated intestinal injury. A selective mineralocorticoid receptor antagonism, eplerenone, inhibited IL17-producing ILC3s and its ROS/RNS production, protected mice from DSS-induced colitis. Mice lacking Nr3c2 (MR coding gene) in ILC3s exhibited decreased IL17 and ROS/RNS production, which alleviated colitis and colitis-associated colorectal cancer (CAC). Further experiments revealed that MR could directly bind to IL17A promoter and facilitate its transcription, which could be enhanced by aldosterone. Thus, our findings demonstrated the critical role of aldosterone-MR-IL17 signaling in ILC3s and gut homeostasis, indicating the therapeutic strategy of eplerenone in IBD clinical trial.


Assuntos
Colite , Doenças Inflamatórias Intestinais , Camundongos , Animais , Receptores de Mineralocorticoides/genética , Receptores de Mineralocorticoides/metabolismo , Aldosterona/metabolismo , Eplerenona , Mineralocorticoides/metabolismo , Imunidade Inata , Espécies Reativas de Oxigênio/metabolismo , Linfócitos , Colite/induzido quimicamente , Colite/tratamento farmacológico , Inflamação/metabolismo
3.
J Bone Oncol ; 41: 100493, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37501717

RESUMO

Osteosarcoma (OS) is the most common primary bone cancer in children and young adults, patient survival rates have not improved in recent decades. To further understand the interrelationship between different cell types in the tumor microenvironment of osteosarcoma, we comprehensively analyzed single-cell sequencing data from six patients with untreated osteosarcoma. Nine major cell types were identified from a total of 46,046 cells based on unbiased clustering of gene expression profiles and canonical markers. Osteosarcoma from different patients display heterogeneity in cellular composition. Myeloid cells were the most commonly represented cell type, followed by osteoblastic and TILs. Copy number variation (CNV) results identified amplifications and deletions in malignant osteoblastic cells and fibroblasts. Trajectory analysis based on RNA velocity showed that osteoclasts in the OS microenvironment could be differentiated from myeloid cells. Furthermore, we explored the intercellular communications in OS microenvironment and identified multiple ligand-receptor pairs between myeloid cells, osteoblastic cells and their cells, including 21 ligand-receptor pair genes that significantly associated with survival outcomes. Importantly, we found chemotherapy may have an effect on cellular communication in the OS microenvironment by analyzing single-cell sequencing data from seven primary osteosarcoma patients who received chemotherapy. We believe these observations will improve our understanding of potential mechanisms of microenvironment contributions to OS progression and help identify potential targets for new treatment development in the future.

4.
Biochem Biophys Res Commun ; 640: 164-172, 2023 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-36512848

RESUMO

Osteoarthritis (OA) places a significant burden on society and finance, and there is presently no effective treatment beside late replacement surgery and symptomatic relief. The therapy of OA requires additional research. Gardenoside is a naturally compound extracted from Gardenia jasminoides Ellis, which has a variety of anti-inflammatory effects. However, few studies have been conducted to determine the role of gardenoside in OA. This study aimed to explore whether gardenoside has effect in OA treatment. Rat primary chondrocytes were treated with IL-1ß to simulate inflammatory environmental conditions and OA in vitro. We examined the effects of gardenoside at concentrations ranging from 0 to 200 µM on the viability of rat chondrocytes and selected 10 µM for further study. Via in vitro experiments, our study found that gardenoside lowers the gene expression of COX-2, iNOS, IL-6, and reduced the ROS production of chondrocytes induced by IL-1ß. Moreover, it effectively alleviates ECM degradation caused by IL-1ß and promotes the ECM synthesis in chondrocytes by upregulating collagen-II and the ACAN expression, downregulating the expression of MMP-3, MMP-13, and ADAMTS-5 expression. Further, our study showed that gardenoside inhibits NF-κB signaling pathway activated by IL-1ß in chondrocytes. We established an OA rat model by anterior cruciate ligament transection (ACLT). The animals were then periodically injected with gardenoside into the knee articular cavity. In vivo study suggested that gardenoside attenuates OA progression in rats. As a whole, in vitro and in vivo results highlight gardenoside is a promising OA treatment agent.


Assuntos
Matriz Extracelular , Iridoides , NF-kappa B , Osteoartrite , Animais , Ratos , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Inflamação/tratamento farmacológico , Interleucina-1beta/metabolismo , Iridoides/farmacologia , Iridoides/uso terapêutico , NF-kappa B/metabolismo , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Transdução de Sinais , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo
5.
Drug Des Devel Ther ; 16: 3793-3804, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36345305

RESUMO

Purpose: Osteoarthritis (OA) places a significant burden on society and finance, and there is presently no effective treatment besides late replacement surgery and symptomatic relief. The therapy of OA requires additional research. Madecassic acid (MA) is the first native triterpenoid compound extracted from Centella asiatica, which has a variety of anti-inflammatory effects. However, the role of MA in OA therapy has not been reported. This study aimed to explore whether MA could suppress the inflammatory response, preserve and restore chondrocyte functions, and ameliorate the progression of OA in vitro and in vivo. Methods: Rat primary chondrocytes were treated with IL-1ß to simulate inflammatory environmental conditions and OA in vitro. We examined the effects of MA at concentrations ranging from 0 to 200 µM on the viability of rat chondrocytes and selected 10 µM for further study. Using qRT-PCR, immunofluorescent, immunocytochemistry, and Western blotting techniques, we identified the potential molecular mechanisms and signaling pathways that are responsible for these effects. We established an OA rat model by anterior cruciate ligament transection (ACLT). The animals were then periodically injected with MA into the knee articular cavity. Results: We found that MA could down-regulate the IL-1ß-induced up-regulation of COX-2, iNOS and IL-6 and restore the cytoskeletal integrity of chondrocytes treated with IL-1ß. Moreover, MA protects chondrocytes from IL-1ß-induced ECM degradation by upregulating ECM synthesis related protein expression, including collagen-II and ACAN, and further down-regulating ECM catabolic related protein expression, including MMP-3 and MMP-13. Furthermore, we found that NF-κB/IκBα and PI3K/AKT signaling pathways were involved in the regulatory effects of MA on the inflammation inhibition and promotion of ECM anabolism on IL-1ß-induced chondrocytes. Conclusion: These findings suggest that MA appears to be a potentially small molecular drug for rat OA.


Assuntos
Osteoartrite , Triterpenos , Ratos , Animais , Fosfatidilinositol 3-Quinases/metabolismo , Células Cultivadas , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Condrócitos , Interleucina-1beta/metabolismo , Triterpenos/uso terapêutico , NF-kappa B/metabolismo , Inflamação/tratamento farmacológico
6.
Oxid Med Cell Longev ; 2022: 4824699, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36193063

RESUMO

Connexin43 (Cx43)-mediated gap junctions are vital in maintaining corneal endothelium homeostasis. Tumor necrosis factor-alpha (TNF-α) is among the most important inflammatory factors which cause corneal endothelial dysfunction in various eye diseases. However, the effect of TNF-α on Cx43-mediated gap junctions of the corneal endothelium remains undefined. In the current research, we determined the effect of TNF-α on gap junction intercellular communication (GJIC) in rabbit corneal endothelium. To evaluate alterations of GJIC, if any, we treated ex vivo cultured rabbit corneal endothelium with different concentrations of TNF-α (2-20 ng/ml). The localization of Cx43 was analyzed by immunostaining, while RT-qPCR and western blot were used to profile the expression of Cx43 and zonula occludens-1 (ZO-1). The association between ZO-1 and Cx43 was evaluated using immunoprecipitation and double staining. GJIC activity was determined by the scrap loading and dye transfer assay (SLDT). Our data demonstrated that a high concentration of TNF-α (10 ng/ml and 20 ng/ml) disrupts the Cx43 mediated gap junction distribution in rabbit corneal endothelium and suppresses the expression of Cx43 protein. Furthermore, rabbit corneal endothelial GJIC was inhibited due to the decreased association between the ZO-1 and Cx43 proteins. Current results demonstrate that TNF-α inhibits corneal endothelial GJIC via decreasing the association between ZO-1 and Cx43, disrupting the distribution of Cx43, and downregulating the expression of Cx43 protein. This study offers a new theoretical foundation for diagnosing and treating corneal endothelial cell decompensation induced by elevated TNF-α in various eye diseases.


Assuntos
Conexina 43 , Fator de Necrose Tumoral alfa , Animais , Comunicação Celular/fisiologia , Células Cultivadas , Junções Comunicantes/metabolismo , Coelhos , Fator de Necrose Tumoral alfa/metabolismo
7.
Mol Neurobiol ; 59(9): 5734-5749, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35781632

RESUMO

Ischemic stroke is a clinical syndrome caused by the disruption of blood flow into cerebral tissues and is associated with high disability and mortality rates. Studies have established the pathological role of platelets in cerebral ischemia/reperfusion (I/R) injury, although the underlying mechanism of action remains largely unclear. In this study, we created an I/R mouse model via middle cerebral artery occlusion and reperfusion (MCAO/R) and analyzed the transcriptomic profiles of the ipsilateral and contralateral cortices using RNA-seq. We found that cerebral I/R injury induced platelet invasion and accumulation in the cerebral cortex by stimulating TNF-α secretion from activated astrocytes in the ischemic region, while TNF-α expression enhanced platelet reactivity through the RIP1/RIP3/AKT pathway. Furthermore, the inoculation of TNF-α-stimulated platelets aggravated I/R injury in mice, whereas the administration of anti-TNF-α antibodies at the onset of reperfusion alleviated ischemic damage. The RNA-seq results further showed that AP-1 transcriptionally activated TNF-α in the I/R-injured cortex by directly binding to the promoter region. These findings provide novel insights into the pathological role of platelets activated by reactive astrocyte-derived TNF-α in cerebral I/R injury.


Assuntos
Isquemia Encefálica , Traumatismo por Reperfusão , Animais , Astrócitos/metabolismo , Isquemia Encefálica/patologia , Proteínas Ativadoras de GTPase , Infarto da Artéria Cerebral Média/patologia , Camundongos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteína Serina-Treonina Quinases de Interação com Receptores , Traumatismo por Reperfusão/patologia , Transdução de Sinais , Inibidores do Fator de Necrose Tumoral , Fator de Necrose Tumoral alfa/farmacologia
8.
Int J Gen Med ; 15: 4635-4647, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35535142

RESUMO

Background: Spindle and kinetochore-associated complex subunit 3 (SKA3) plays important roles in promoting the migration and the invasion of various human cancer cells. There are a few studies on SKA3 in lung adenocarcinoma (LUAD), but the in-depth analysis of the expression of SKA3 and the correlated possible immune mechanism of SKA3 in LUAD are not clear. Methods: In our study, the expression and survival data of SKA3 were analyzed in LUAD using TIMER, Oncomine, UALCAN, cBioPortal, LinkedOmics, Human Protein Atlas, and Kaplan-Meier plotter. Then, quantitative PCR was used to verify the expression differences of SKA3 between LUAD tissues of mice and the normal tissues. Results: We established that the expression of SKA3 in the LUAD group was remarkably higher than that in the normal group. Additionally, high SKA3 expression was linked to poorer survival in LUAD. Moreover, SKA3 expression had a remarkable negative correlation with the immune infiltration of B cells, macrophages, and CD4+ T cells. SKA3 was markedly negatively related to the immune type biomarkers of T cells and B cells in LUAD. The elevated expression of SKA3 with LUAD in enriched B cells, CD4+ T cells, CD8+ T cells, macrophages and Treg cells had worse prognosis, respectively. Functional network analysis showed that SKA3 regulated the mitotic cell cycle, mitosis, chromosome segregation and cell division via pathways. Conclusion: In summary, our study suggested that SKA3 was highly expressed in LUAD and SKA3 might function as a prognostic biomarker in LUAD. Besides, SKA3 may be a candidate oncogene, which correlates with poor prognosis and immune infiltration in lung adenocarcinoma.

9.
Phytomedicine ; 100: 154089, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35398736

RESUMO

BACKGROUND: Rabdosia japonica (Burm. f.) var. glaucocalyx (Maxim.) is a perennial herb, and is traditionally used as folk medicine for treating inflammatory diseases and cancer. Gaucocalyxin A (GLA) is an ent­kaurane diterpenoid that is isolated from the aerial parts of R. japonica (Burm. f.) var. glaucocalyx (Maxim.). In a recent study, we found that GLA protects against acute liver dysfunction induced by Escherichia coli, which is likely related to its anti-inflammatory effects. However, the mechanism by which GLA protects liver injury during sepsis is unknown. AIM: To evaluate the anti-inflammatory function of GLA and its regulatory effect on platelet function. METHOD: An in vivo model of sepsis was established by inoculating mice with E. coli. Live function and platelet activation were evaluated through standard assays. The levels of pro-inflammatory factors were measured through ELISA and qRT-PCR. RESULTS: GLA alleviated liver dysfunction in the mouse model of sepsis. GLA-treated mice displayed lower complement activation and liver dysfunction after E. coli infection. GLA alleviated the decrease in peripheral platelet counts by inhibiting their clearance by Kupffer cells in liver. Furthermore, GLA inhibited platelet activation through the RIP1/RIP3/AKT pathway and downregulated C3aR expression on the platelets, thereby inhibiting liver injury and dysfunction due to excessive complement activation. CONCLUSION: GLA can inhibit platelet activation by reducing surface expression of C3aR, which protect the liver from injury induced by excessive complement activation. GLA is a novel therapeutic agent for controlling sepsis-related liver dysfunction.


Assuntos
Diterpenos do Tipo Caurano , Sepse , Animais , Diterpenos do Tipo Caurano/farmacologia , Escherichia coli , Fígado , Camundongos , Ativação Plaquetária , Sepse/tratamento farmacológico , Transdução de Sinais
10.
Am J Physiol Renal Physiol ; 320(3): F505-F517, 2021 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33522410

RESUMO

Recent evidence suggests that dipeptidyl peptidase-4 (DPP4) inhibition with saxagliptin (Saxa) is renoprotective under comorbid conditions associated with activation of the renin-angiotensin-aldosterone system (RAAS), such as diabetes, obesity, and hypertension, which confer a high cardiovascular risk. Immune system activation is now recognized as a contributor to RAAS-mediated tissue injury, and, importantly, immunomodulatory effects of DPP4 have been reported. Accordingly, we examined the hypothesis that DPP4 inhibition with Saxa attenuates angiotensin II (ANG II)-induced kidney injury and albuminuria via attenuation of immune activation in the kidney. To this end, male mice were infused with either vehicle or ANG II (1,000 ng/kg/min, s.c.) for 3 wk and received either placebo or Saxa (10 mg/kg/day, p.o.) during the final 2 wk. ANG II infusion increased kidney, but not plasma, DPP4 activity in vivo as well as DPP4 activity in cultured proximal tubule cells. The latter was prevented by angiotensin receptor blockade with olmesartan. Further, ANG II induced hypertension and kidney injury characterized by mesangial expansion, mitochondrial damage, reduced brush border megalin expression, and albuminuria. Saxa inhibited DPP4 activity ∼50% in vivo and attenuated ANG II-mediated kidney injury, independent of blood pressure. Further mechanistic experiments revealed mitigation by Saxa of proinflammatory and profibrotic mediators activated by ANG II in the kidney, including CD8+ T cells, resident macrophages (CD11bhiF4/80loLy6C-), and neutrophils. In addition, Saxa improved ANG II suppressed anti-inflammatory regulatory T cell and T helper 2 lymphocyte activity. Taken together, these results demonstrate, for the first time, blood pressure-independent involvement of renal DPP4 activation contributing to RAAS-dependent kidney injury and immune activation.NEW & NOTEWORTHY This work highlights the role of dipeptidyl peptidase-4 (DPP4) in promoting ANG II-mediated kidney inflammation and injury. Specifically, ANG II infusion in mice led to increases in blood pressure and kidney DPP4 activity, which then led to activation of CD8+ T cells, Ly6C- macrophages, and neutrophils and suppression of anti-inflammatory T helper 2 lymphocytes and regulatory T cells. Collectively, this led to kidney injury, characterized by mesangial expansion, mitochondrial damage, and albuminuria, which were mitigated by DPP4 inhibition independent of blood pressure reduction.


Assuntos
Dipeptidil Peptidase 4/metabolismo , Hipoglicemiantes/farmacologia , Macrófagos/metabolismo , Angiotensina II/farmacologia , Animais , Inibidores da Dipeptidil Peptidase IV/farmacologia , Hipertensão/tratamento farmacológico , Hipertensão/fisiopatologia , Rim/efeitos dos fármacos , Rim/metabolismo , Túbulos Renais Proximais/efeitos dos fármacos , Túbulos Renais Proximais/metabolismo , Macrófagos/efeitos dos fármacos , Masculino , Camundongos
11.
Asia Pac J Clin Nutr ; 29(2): 234-238, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32674229

RESUMO

BACKGROUND AND OBJECTIVES: We investigated the effects of a carbohydrate-whey protein solution on aged patients undergoing hip fracture surgery. METHODS AND STUDY DESIGN: Forty patients were randomly assigned to the carbohydrate-whey protein (CHP) group or the control group (CTL). In the CHP group, a mixed solution of CHP was orally administered to patients before surgery: 400 mL was administered on the day before surgery, and 200 mL was administered 3 h before surgery. The size of the liquid dark area in the gastric antrum was measured by ultrasound, and the bleeding volume during surgery was assayed. The incidence of nausea, vomiting, thirst, hunger, and days of hospitalization and the levels of blood glucose, C-reactive protein (CRP) and serum albumin were assessed. RESULTS: There was no obvious liquid dark space in the gastric antrum. CHP administration improved postoperative thirst and hunger and resulted in increased albumin levels and decreased CRP concentrations and blood glucose fluctuations. CONCLUSIONS: Oral CHP before hip fracture surgery reduces the incidence of postoperative thirst and hunger and improves recovery in the aged.


Assuntos
Bebidas , Idoso Fragilizado , Fraturas do Quadril/cirurgia , Náusea/prevenção & controle , Idoso de 80 Anos ou mais , Feminino , Glucose , Humanos , Masculino , Complicações Pós-Operatórias/prevenção & controle , Período Pré-Operatório , Resultado do Tratamento , Proteínas do Soro do Leite
12.
Exp Ther Med ; 18(6): 4935-4941, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31798715

RESUMO

Hypothyroidism is associated with profound left ventricular dysfunction. Triiodothyronine (T3) supplementation may improve cardiac function after ischemic reperfusion (I/R) injury. In the present study, the effect of T3 on major calcium cycling proteins and high-energy phosphate content during I/R was evaluated. Isolated perfused rat hearts were divided into 5 groups: Sham Control (Sham, n=10), Control (n=8), T3 10 nM (T3-10, n=10), T3 25 nM (T3-25, n=10) and T3 50 nM (T3-50, n=10). T3 was administrated for 60 min before 30 min of ischemia and 120 min of reperfusion. The protein contents of Ca2+-release channels (RyR2), Ca2+-adenosine triphosphatase (SERCA2a), phospholamban (PLB), sarcolemmal Ca2+-adenosine triphosphatase (PMCA) and sodium-calcium exchanger (NCX), as well as the high-energy phosphate content in heart tissues were measured by western blot analysis. The results revealed that T3 improved the contractile recovery (left ventricular developed pressure; +dP/dt, -dP/dt) after I/R. Western blotting assays demonstrated that I/R depressed the contents of RYR2, SERCA2a and phosphorylated RYR2 and PLB; there were no effects on the contents of PLB, PMCA and NCX. T3 reversed I/R-induced degradation of RyR2 and SERCA2a, restored the phosphorylation of RyR2 and PLB, and preserved the high-energy phosphate contents of ATP and creatine phosphate. T3 supplementation protected the heart against I/R injury via the preservation of Ca2+-cycling proteins and high-energy phosphate content.

13.
Cardiorenal Med ; 8(2): 113-122, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29617003

RESUMO

BACKGROUND: Anesthetic preconditioning (APC) of the myocardium is mediated in part by reversible alteration of mitochondrial function. Nitric oxide (NO) inhibits mitochondrial respiration and may mediate APC-induced cardioprotection. In this study, the effects of isoflurane on different states of mitochondrial respiration during the oxidation of complex I-linked substrates and the role of NO were investigated. METHODS: Mitochondria were isolated from Sprague-Dawley rat hearts. Respiration rates were measured polarographically at 28ºC with a computer-controlled Clark-type O2 electrode in the mitochondria (0.5 mg/mL) with complex I substrates glutamate/malate (5 mM). Isoflurane (0.25 mM) was administered before or after adenosine diphosphate (ADP)-initiated state 3 respiration. The NO synthase (NOS) inhibitor L-N5-(1-iminoethyl)-ornithine (L-NIO, 10 µM) and the NO donor S-nitroso-N-acetylpenicillamine (SNAP, 1 µM) were added before or after the addition of ADP. RESULTS: Isoflurane administered in state 2 increased state 2 respiration and decreased state 3 respiration. This attenuation of state 3 respiration by isoflurane was similar when it was given during state 3. L-NIO did not alter mitochondrial respiration or the effect of isoflurane. SNAP only, added in state 3, decreased state 3 respiration and enhanced the isoflurane-induced attenuation of state 3 respiration. CONCLUSION: Isoflurane has clearly distinguishable effects on different states of mitochondrial respiration during the oxidation of complex I substrates. The uncoupling effect during state 2 respiration and the attenuation of state 3 respiration may contribute to the mechanism of APC-induced cardioprotection. These effects of isoflurane do not depend on endogenous mitochondrial NO, as the NOS inhibitor L-NIO did not alter the effects of isoflurane on mitochondrial respiration.


Assuntos
Precondicionamento Isquêmico Miocárdico/métodos , Isoflurano/farmacologia , Mitocôndrias Cardíacas/metabolismo , Isquemia Miocárdica/metabolismo , Miocárdio/metabolismo , Óxido Nítrico/biossíntese , Anestésicos Inalatórios/farmacologia , Animais , Modelos Animais de Doenças , Masculino , Mitocôndrias Cardíacas/efeitos dos fármacos , Isquemia Miocárdica/patologia , Oxirredução , Ratos , Ratos Sprague-Dawley
14.
Shock ; 37(3): 325-32, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22089203

RESUMO

Recently, we reported that Brown Norway (BN) rats were more resistant to lipopolysaccharide (LPS)-induced myocardial dysfunction than Dahl S (SS) rats. This differential sensitivity was exemplified by reduced production of proinflammatory cytokines and diminished nuclear factor-κB pathway activation. To further clarify the mechanisms of different susceptibility of these two strains to endotoxin, this study was designed to examine the alterations of cardiac and mitochondrial bioenergetics, proinflammatory cytokines, and signaling pathways after hearts were isolated and exposed to LPS ex vivo. Isolated BN and SS hearts were perfused with LPS (4 µg/mL) for 30 min in the Langendorff preparation. Lipopolysaccharide depressed cardiac function as evident by reduced left ventricular developed pressure and decreased peak rate of contraction and relaxation in SS hearts but not in BN hearts. These findings are consistent with our previous in-vivo data. Under complex I substrates, a higher oxygen consumption and hydrogen peroxide (H2O2) production were observed in mitochondria from SS hearts than those from BN hearts. Lipopolysaccharide significantly increased H2O2 levels in both SS and BN heart mitochondria; however, the increase in oxygen consumption and H2O2 production in BN heart mitochondria was much lower than that in SS heart mitochondria. In addition, LPS significantly decreased complex I activity in SS hearts but not in BN hearts. Furthermore, LPS induced higher levels of tumor necrosis factor-α and increased phosphorylation of IκκB and p65 more in SS hearts than in BN hearts. Our results clearly demonstrate that less mitochondrial dysfunction combined with a reduced production of tumor necrosis factor-α and diminished activation of nuclear factor-κB are involved in the mechanisms by which isolated BN hearts were more resistant to LPS-induced myocardial dysfunction.


Assuntos
Insuficiência Cardíaca/induzido quimicamente , Coração/efeitos dos fármacos , Lipopolissacarídeos , Mitocôndrias Cardíacas/fisiologia , Miocárdio/metabolismo , NF-kappa B/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Animais , Complexo I de Transporte de Elétrons/efeitos dos fármacos , Complexo II de Transporte de Elétrons/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Técnicas In Vitro , Interleucina-1beta/biossíntese , Interleucina-6/biossíntese , Lipopolissacarídeos/farmacologia , Masculino , Mitocôndrias Cardíacas/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Contração Miocárdica/efeitos dos fármacos , Consumo de Oxigênio , Perfusão , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos Dahl
15.
Shock ; 33(3): 332-6, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20160611

RESUMO

We previously demonstrated that hearts from Brown Norway (BN) rats were more resistant to ischemic injury than hearts from Dahl S (SS) rats. Here we determined the susceptibility to LPS-induced cardiomyopathy in these rats and examined the involvement of inflammatory signaling. Both strains were treated with LPS (20 mg/kg) via i.p. injection for 6 h. Myocardial function was assessed by the Langendorff system, and proinflammatory cytokines were measured by the enzyme-linked immunosorbent assay. LPS significantly reduced left ventricular developed pressure in both strains. Interestingly, the decrease of left ventricular developed pressure in BN rat hearts was approximately 25% less than that in SS rat hearts. Furthermore, LPS significantly reduced the peak rate of contraction and the peak rate of relaxation in SS hearts but not in BN hearts. No differences in LPS-induced decreases in coronary flow rate were observed between BN and SS rats. In addition, LPS-induced increases in proinflammatory cytokines, TNF-alpha, IL-1beta, and IL-6, were significantly lower in both plasma and hearts of BN rats compared with production in SS rats. LPS notably up-regulated the expression of proinflammatory enzymes, iNOS and cyclooxygenase 2, in SS hearts but not in BN hearts. Interestingly, LPS did not stimulate Toll-like receptor 4 or its adaptor myeloid differentiation factor 88 expression in the hearts of either strain but did increase IkappaB and P65 phosphorylation, less prominently in BN hearts than in SS hearts. These data indicate that reduced production of proinflammatory cytokines and diminished nuclear factor kappaB activation are major mechanisms by which BN hearts are more resistant to LPS-induced myocardial dysfunction than SS hearts.


Assuntos
Cardiomiopatias/induzido quimicamente , NF-kappa B/fisiologia , Ratos Endogâmicos BN/metabolismo , Ratos Endogâmicos Dahl/metabolismo , Animais , Cardiomiopatias/metabolismo , Ciclo-Oxigenase 2/biossíntese , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos , Masculino , Contração Miocárdica/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/biossíntese , Ratos , Fator de Necrose Tumoral alfa/metabolismo
16.
Am J Physiol Cell Physiol ; 297(3): C742-9, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19553566

RESUMO

In inflammatory diseases, tissue damage is critically associated with nitric oxide ((*)NO) and cytokines, which are overproduced in response to cellular release of endotoxins. Here we investigated the inhibitory effect of roscovitine, a selective inhibitor of cyclin-dependent kinases (CDKs) on (*)NO production in mouse macrophages. In RAW264.7 cells, we found that roscovitine abolished the production of (*)NO induced by lipopolysaccharide (LPS). Moreover, roscovitine significantly inhibited LPS-induced inducible nitric oxide synthase (iNOS) mRNA and protein expression. Our data also showed that roscovitine attenuated LPS-induced phosphorylation of IkappaB kinase beta (IKKbeta), IkappaB, and p65 but enhanced the phosphorylation of ERK, p38, and c-Jun NH(2)-terminal kinase (JNK). In addition, roscovitine dose dependently inhibited LPS-induced expression of cyclooxygenase-2 (COX)-2, IL-1beta, and IL-6 but not tumor necrosis factor (TNF)-alpha. Tetrahydrobiopterin (BH(4)), an essential cofactor for iNOS, is easily oxidized to 7,8-dihydrobiopterin (BH(2)). Roscovitine significantly inhibited LPS-induced BH(4) biosynthesis and decreased BH(4)-to-BH(2) ratio. Furthermore, roscovitine greatly reduced the upregulation of GTP cyclohydrolase-1 (GCH-1), the rate-limiting enzyme for BH(4) biosynthesis. Using other CDK inhibitors, we found that CDK1, CDK5, and CDK7, but not CDK2, significantly inhibited LPS-induced (*)NO production in macrophages. Similarly, in isolated peritoneal macrophages, roscovitine strongly inhibited (*)NO production, iNOS, and COX-2 upregulation, activation of NFkappaB, and induction of GCH-1 by LPS. Together, our data indicate that roscovitine abolishes LPS-induced (*)NO production in macrophages by suppressing nuclear factor-kappaB activation and BH(4) biosynthesis, which might be mediated by CDK1, CDK5, and CDK7. Our results also suggest that roscovitine may inhibit inflammation and that CDKs may play important roles in the mechanisms by which roscovitine attenuates inflammation.


Assuntos
Biopterinas/análogos & derivados , Quinases Ciclina-Dependentes/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , NF-kappa B/metabolismo , Óxido Nítrico/biossíntese , Purinas/farmacologia , Animais , Biopterinas/biossíntese , Linhagem Celular , Quinases Ciclina-Dependentes/metabolismo , Relação Dose-Resposta a Droga , GTP Cicloidrolase/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico Sintase Tipo II/metabolismo , Roscovitina
17.
Anesth Analg ; 105(5): 1207-13, table of contents, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17959942

RESUMO

INTRODUCTION: Anesthetic preconditioning (APC) with volatile anesthetics improves recovery of contractile function and reduces calcium overload after ischemia/reperfusion (I/R). Mitochondrial and sarcolemmal K(ATP) channel openings have been implicated in APC-induced cardioprotection. In this study, we investigated the effect of APC on major calcium cycling proteins and its relation to K(ATP) channels. METHODS: Isolated perfused rat hearts were divided into seven groups: Time control (n = 10), ischemia control (n = 8), APC (n = 8), Mitochondrial K(ATP) inhibitor 5-hydroxydecanoate (5-HD, 200 microM, n = 8), Sarcolemmal K(ATP) inhibitor HMR1098 (HMR, 20 microM, n = 8), and APC plus 5-HD or APC plus HMR1098 (n = 8 each). APC was initiated by administering 1.5% isoflurane for 15 min, followed by a 15 min washout before 30 min of myocardial ischemia and 60 min of reperfusion. Ca2+-release channels (RyR2), Ca2+-adenosine triphosphatase (SERCA2a), phospholamban, plasma membrane Ca2+ ATPase, and sodium-calcium exchanger in the homogenate were determined by Western blot assay. RESULTS: APC improved contractile recovery (left ventricular developed pressure, +dP/dt, -dP/dt) after I/R, which was blocked by 5-HD and HMR. I/R depressed the density of RyR2, SERCA2a, and phospholamban, with no changes in the density of plasma membrane Ca2+ ATPase and sodium-calcium exchanger. APC reversed I/R-induced degradation of RyR2 and SERCA2a in the presence or absence of 5HD and HMR. CONCLUSIONS: I/R-induced depression in cardiac performance is associated with a down-regulation of the major sarcoplasmic reticulum Ca2+-cycling proteins. Anesthesia preconditioning with isoflurane prevents I/R-related degradation of the RyR2 and SERCA2a in the sarcoplasmic reticulum. However, this effect was independent of its activation of K(ATP) channels.


Assuntos
Cálcio/metabolismo , Precondicionamento Isquêmico Miocárdico/métodos , Isoflurano/uso terapêutico , Mitocôndrias Cardíacas/metabolismo , Canais de Potássio/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Sarcolema/metabolismo , Animais , Técnicas In Vitro , Masculino , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/prevenção & controle , Ratos , Ratos Wistar , Canal de Liberação de Cálcio do Receptor de Rianodina/análise
18.
Anesthesiology ; 105(3): 541-9, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16931987

RESUMO

BACKGROUND: Anesthetic preconditioning (APC) is well known to protect against myocardial ischemia-reperfusion injury. Studies also show the benefit of Na+-Ca2+ exchange inhibition on ischemia-reperfusion injury. The authors tested whether APC plus Na+-Ca2+ exchange inhibitors given just on reperfusion affords additive protection in intact hearts. METHODS: Cytosolic [Ca2+] was measured by fluorescence at the left ventricular wall of guinea pig isolated hearts using indo-1 dye. Sarcoplasmic reticular Ca2+-cycling proteins, i.e., Ca2+ release channel (ryanodine receptor [RyR2]), sarcoplasmic reticular Ca2+-pump adenosine triphosphatase (SERCA2a), and phospholamban were measured by Western blots. Hearts were assigned to seven groups (n = 8 each): (1) time control; (2) ischemia; (3, 4) 10 microM Na+-Ca2+ exchange inhibitor KB-R7943 (KBR) or 1 microM SEA0400 (SEA), given during the first 10 min of reperfusion; (5) APC initiated by sevoflurane (2.2%, 0.41 +/- 0.03 mm) given for 15 min and washed out for 15 min before ischemia-reperfusion; (6, 7) APC plus KBR or SEA. RESULTS: The authors found that APC reduced the increase in systolic [Ca2+], whereas KBR and SEA both reduced the increase in diastolic [Ca2+] on reperfusion. Each intervention improved recovery of left ventricular function. Moreover, APC plus KBR or SEA afforded better functional recovery than APC, KBR, or SEA alone (P < 0.05). Ischemia-reperfusion-induced degradation of major sarcoplasmic reticular Ca2+-cycling proteins was attenuated by APC, but not by KBR or SEA. CONCLUSIONS: APC plus Na+-Ca2+ exchange inhibition exerts additive protection in part by reducing systolic and diastolic Ca2+ overload, respectively, during ischemia-reperfusion. Less degradation of sarcoplasmic reticular Ca2+-cycling proteins may also contribute to cardiac protection.


Assuntos
Anestésicos/farmacologia , Compostos de Anilina/farmacologia , Cálcio/metabolismo , Precondicionamento Isquêmico Miocárdico , Miocárdio/metabolismo , Éteres Fenílicos/farmacologia , Trocador de Sódio e Cálcio/antagonistas & inibidores , Tioureia/análogos & derivados , Animais , ATPases Transportadoras de Cálcio/antagonistas & inibidores , Cobaias , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático , Tioureia/farmacologia
19.
Anesthesiology ; 100(3): 575-80, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15108971

RESUMO

BACKGROUND: Myocardial protection by volatile anesthetics involves activation of cardiac adenosine triphosphate-sensitive potassium (K(ATP)) channels. The authors have previously shown that isoflurane enhances sensitivity of the sarcolemmal K(ATP) channel to the opener, pinacidil. Because reactive oxygen species seem to be mediators in anesthetic preconditioning, the authors investigated whether they contribute to the mechanism of the sensitization effect by isoflurane. METHODS: Ventricular myocytes were isolated from guinea pig hearts for the whole cell patch clamp recordings of the sarcolemmal K(ATP) channel current (I(KAPT)). Free radical scavengers N-acetyl-L-cysteine, carnosine, superoxide dismutase, and catalase were used to investigate whether reactive oxygen species mediate isoflurane facilitation of the channel opening by pinacidil. A possible role of the mitochondrial K(ATP) channels was tested using a blocker of these channels, 5-hydroxydecanoate. RESULTS: The mean density (+/- SEM) of I(KAPT) elicited by pinacidil (20 microM) was 18.9 +/- 1.8 pA/pF (n = 11). In the presence of isoflurane (0.55 mM), the density of pinacidil-activated I(KAPT) increased to 38.5 +/- 2.4 pA/pF (n = 9). Concurrent application of isoflurane and N-acetyl-L-cysteine decreased the sensitization effect by isoflurane in a concentration-dependent manner, whereby the densities of I(KAPT) were 32.6 +/- 1.4 (n = 6), 26.2 +/- 2.3 (n = 6), and 19.4 +/- 2.1 pA/pF (n = 8) at 100, 250, and 500 microM N-acetyl-L-cysteine, respectively. Concurrent application of isoflurane and carnosine (100 microM), superoxide dismutase (100 U/ml), or catalase (100 U/ml) attenuated the densities of I(KAPT) to 27.9 +/- 2.6, 27.2 +/- 2.9, and 25.9 +/- 2.2 pA/pF, respectively. None of the scavengers affected activation of I(KAPT) by pinacidil alone. 5-Hydroxydecanoate (100 microM) did not alter the sensitization effect by isoflurane, and the density of I(KAPT) in this group was 37.1 +/- 3.8 pA/pF (n= 6). CONCLUSION: These results suggest that reactive oxygen species contribute to the mechanism by which isoflurane sensitizes the cardiac sarcolemmal K(ATP) channel to the opener, pinacidil.


Assuntos
Anestésicos Inalatórios/farmacologia , Coração/efeitos dos fármacos , Isoflurano/farmacologia , Proteínas de Membrana/efeitos dos fármacos , Pinacidil/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Sarcolema/metabolismo , Vasodilatadores/farmacologia , Acetilcisteína/farmacologia , Animais , Carnosina/farmacologia , Catalase/metabolismo , Separação Celular , Cobaias , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Proteínas de Membrana/agonistas , Miocárdio/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Técnicas de Patch-Clamp , Canais de Potássio , Sarcolema/efeitos dos fármacos , Superóxido Dismutase/metabolismo
20.
Eur J Cardiothorac Surg ; 24(6): 974-85, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14643817

RESUMO

OBJECTIVE: Cold cardioplegia (CP) protects against ischemic damage in part by reducing [Ca(2+)](i) overload on reperfusion. Hyperkalemic cardioplegic solutions are widely used in coronary artery bypass procedures, and the specific ionic composition of these solutions may contribute to their variable myocardial protective effects secondary to reduced Ca(2+)(i) loading. We reported previously that CP decreased the rise in cardiac diastolic (dia) [Ca(2+)](i) observed during 4 h cold storage at 3 degrees C in Krebs-Ringer's (KR) solution and decreased dia[Ca(2+)](i) and increased systolic (sys) [Ca(2+)](i) and function on reperfusion after cold storage. Our aim here was to determine if low Ca(2+)(o) and high Mg(2+)(o) adds to the protective effects of high K(+)(o) by decreasing [Ca(2+)](i) during ischemia and reperfusion. METHODS: We compared effects of 4.5 mM K(+)(o), 2.5 mM Ca(2+)(o) and 2.4 mM Mg(2+)(o) KR solution with a higher K(+)(o) (18 mM), a lower Ca(2+)(o) (1.25 mM) and/or higher Mg(2+)(o) (7.2 mM) CP solutions on cardiac mechanic function and sys and dia[Ca(2+)](i) during and after moderate hypothermic global ischemia (17 degrees C for 4 h) in guinea pig intact hearts isolated by the Langendorff technique. Isovolumetric left ventricular pressure (LVP) was measured with a transducer connected to a fluid-filled balloon placed in the LV and [Ca(2+)](i) was measured using indo-1 fluorescence and a fiberoptic cable placed on the LV free wall. RESULTS: For all CP groups compared to the KR control group after 60 min reperfusion, we observed significant lowering of dia[Ca(2+)](i) by 47%, left ventricular diastolic pressure (diaLVP) by 55%, and infarct size by 43%. We also found significant elevation of sys[Ca(2+)](i) by 25%, d[Ca(2+)](i)/dt(max) and d[Ca(2+)](i)/dt(min) by 33 and 34%, sys-diaLVP by 55%, dLVP/dt(max) and dLVP/dt(min) by 34 and 40%, coronary flow by 31%, cardiac efficiency by 21%, and MVO(2) by 25%. These results indicate that CP reduces myoplasmic Ca(2+) loading and improves mechanical and metabolic function on warm reperfusion compared to KR. However, there were no differences in these indices of Ca(2+)(i) cardiac function or metabolism among any CP group after warm reperfusion with KR solution. CONCLUSION: Increasing K(+)(o) to produce cardiac arrest was the most cardioprotective effect of CP against ischemia reperfusion injury; lowering Ca(2+)(o) or raising Mg(2+)(o) did not add to this protective effect or additionally alter [Ca(2+)](i).


Assuntos
Cloreto de Cálcio/farmacologia , Soluções Cardioplégicas/farmacologia , Parada Cardíaca Induzida/métodos , Coração/efeitos dos fármacos , Cloreto de Magnésio/farmacologia , Animais , Cálcio/metabolismo , Cobaias , Traumatismo por Reperfusão Miocárdica/patologia , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/metabolismo , Técnicas de Cultura de Órgãos , Potássio/farmacologia , Função Ventricular Esquerda/efeitos dos fármacos
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