Intrathecal betamethasone pain relief in cancer patients with vertebral metastasis: a pilot study

BACKGROUND: We have reported previously the usefulness of intrathecal betamethasone for pain relief in cancer patients who suffer from intractable pain caused by vertebral metastasis. The mechanism by which betamethasone relieves pain may be related to alterations in cerebrospinal fluid (CSF) concentrations of pro-inflammatory cytokines and prostanoids. METHODS: Thirteen cancer patients with intractable pain caused by vertebral metastasis received 2-3 mg betamethasone in the lumbar subarachnoid space. CSF concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-6, IL-8 and prostaglandin E(2) (PGE(2)) were measured with an enzyme-linked immunosorbent assay (ELISA) and a chemiluminescence enzyme immunoassay. Pain was measured using a numerical pain score (range, 0-10; 0, no pain; 10, worst pain imaginable). RESULTS: Intrathecal betamethasone was associated with a significant decrease in the pain score in six patients. In these cases, the pain score decreased from 6.7 +/- 0.5 (mean +/- standard error of the mean) to 3.3 +/- 0.3 (P < 0.05), and the CSF concentrations of IL-8 and PGE(2) decreased significantly compared with pre-treatment levels (IL-8, 183.3 +/- 21.2 to 116.5 +/- 10.6 pg/ml; PGE(2), 43.8 +/- 10.3 to 14.7 +/- 3.0 pg/ml). There were no significant changes in the CSF concentrations of cytokines and PGE(2) in the remaining seven patients. CONCLUSION: Pain relief with intrathecal betamethasone is related to decreases in the CSF concentration of IL-8 and PGE(2).     

Effects of short-term inhaled nitric oxide on interleukin-8 release after single-lung transplantation in pigs.

BACKGROUND: Lung transplantation has evolved to become an effective treatment for a variety of end-stage lung diseases. However, severe reperfusion injury is still a major cause for postoperative morbidity and mortality. Although lung reperfusion injury is complex and has not been fully comprehended yet, neutrophil infiltration and cytokine activation have been postulated to play a main role. Recent studies showed that nitric oxide (NO) therapy has salutary effects on lung chronic and acute pathologies because it inhibits interleukin-8 (IL-8) release, but no data have been found on its effects during organ harvest. The aim of this study was to assess whether low doses of inhaled NO pre-treatment at the time of harvesting improves allograft function during early reperfusion in a porcine model. METHODS: Twenty-two Landrace pigs were randomly assigned to NO-treated and control groups. In NO-treated pigs, NO at 20 ppm was administered 30 min before harvest. During the early allograft reperfusion period IL-8 content, dynamic and static compliance and gas exchange (Pa/FiO2 and PaO2) were measured in both control and NO-treated lungs. RESULTS: Pre-treatment with NO at the time of harvesting showed improvement of allograft function in terms of dynamic (92 +/- 8% in NO vs 72 +/- 7% in the control group, p < .05) and static (83 +/- 8% in NO vs 63 +/- 7% in the control group, p < 0.05) compliance and gas exchange (PaO2: 96 +/- 4% in NO vs 74 +/- 4.5% in the control group, p < 0.01; Pa/FiO2: 97 +/- 5% in NO vs 74 +/- 5% in the control group, p < 0.01) by diminishing IL-8 (66.5 +/- 4.7 pg/ml in NO versus 208 +/- 43 pg/ml in the control group, p < 0.05) release in pigs. CONCLUSION: These results show for the first time that NO pre-treatment at the time of harvesting reduces allograft reperfusion injury in part due to its effects on IL-8 release.     

Inducible nitric oxide production is an adaptation to cardiopulmonary bypass-induced inflammatory response

BACKGROUND: Cardiopulmonary bypass (CPB) increases nitric oxide (NO) production by the activation of NO synthases (NOS). However, the role of NO from inducible NOS (iNOS) in CPB-induced inflammatory response remains unclear. We examined the effect of a selective iNOS inhibitor, aminoguanidine, on CPB-induced inflammatory response in a rat-CPB model. METHODS: Adult Sprague-Dawley rats underwent 60 minutes of CPB (100 mL x kg(-1) x min(-1), 34 degrees C). Group A (n = 10) received 100 mg/kg of aminoguanidine intraperitoneally 30 minutes before the initiation of CPB, and group B (n = 10) served as controls. RESULTS: There were significant time-dependent changes in plasma interleukin (IL)-6, IL-8, nitrate + nitrite, the percentage ratio of nitrotyrosine to tyrosine (%NO2-Tyr, an indicator of peroxynitrite formation), and respiratory index (RI). Three hours after CPB termination, IL-6, IL-8, and RI were significantly higher in group A (IL-6, 397.5+/-80.6 pg/mL; IL-8, 26.99+/-6.57 ng/mL; RI, 1.87+/-0.31) than in group B (IL-6, 316.5+/-73.9 pg/mL, p <0.05; IL-8, 17.21+/-3.12 ng/mL, p < 0.01; RI, 1.57+/-0.24, p < 0.05) although nitrate + nitrite (31.8+/-4.1 micromol/L) and %NO2-Tyr (1.15%+/-0.20%) were significantly lower in group A than in group B (nitrate + nitrite, 50.2+/-5.0 micromol/L, p < 0.01; %NO2-Tyr, 1.46%+/-0.21%, p < 0.01). Western immunoblot analysis from lung tissue of group A identified marked iNOS inhibition without inhibiting endothelial-constitutive NOS, and neutrophil accumulation in the lung specimens was significantly greater in group A (6.5+/-0.7/alveoli) than in group B (4.4+/-0.4/alveoli, p < 0.01). CONCLUSIONS: These results suggest that NO production from iNOS may be an adaptive response for attenuating the CPB-induced inflammatory response.     

Proinflammatory cytokines in cerebrospinal fluid in repair of thoracoabdominal aorta

BACKGROUND: Little is known about alterations of cytokine levels in cerebrospinal fluid (CSF) during thoracoabdominal aortic surgery. We measured perioperative CSF cytokine levels to determine their clinical significances. METHODS: Perioperative serum and CSF levels of cytokine were measured in 15 adult patients undergoing repair of the descending thoracic aorta (n = 4) or thoracoabdominal aorta (n = 11). All patients underwent prosthetic replacement and perioperative CSF drainage. Serum and CSF levels of tumor necrosis factor-alpha, Interleukin- (IL-) 1beta, IL-6, IL-8, IL-10, and IL-12 were measured before operation and at 0, 6, 12, 18, 24, 48, and 72 hours postoperatively using enzyme-linked immunosorbent assays. RESULTS: There were no hospital deaths, but 1 patient suffered paraplegia. Cerebrospinal fluid IL-8 levels peaked at immediately after operation (751.7 +/- 42.1 pg/mL versus preoperative levels, 54.9 +/- 24.6 pg/mL; p < 0.001), and the higher levels persisted for 72 hours. In contrast, serum IL-8 levels did not change and remained lower than CSF levels. The patient with paraplegia had the highest CSF IL-8 levels throughout the study period. Serum and CSF levels of tumor necrosis factor-alpha, IL-1beta, IL-6, and IL-12 did not significantly change. Serum and CSF levels of IL-10 were significantly elevated after operation compared with preoperative levels. In contrast to IL-8, serum IL-10 levels surpassed CSF levels. CONCLUSIONS: Cerebrospinal fluid IL-8 levels are significantly elevated in thoracoabdominal aortic operation, and may be the most sensitive to the inflammatory response in the ischemic spinal cord injury. Persistent elevation of CSF IL-8 levels may be predictive of further development of neurologic deficits, and a reduction of proinflammatory cytokine levels may be a beneficial effect of CSF drainage, but this requires further investigation.     

舒芬太尼、瑞芬太尼或芬太尼对食管癌根治术病人细胞免疫功能的影响

目的 探讨舒芬太尼、瑞芬太尼或芬太尼对食管癌根治术病人细胞免疫功能的影响.方法 择期行食管中、下段癌根治术病人45例,ASA Ⅰ或Ⅱ级,年龄45～64岁.随机分为3组(n=15):舒芬太尼组(SF组)、瑞芬太尼组(RF组)和芬太尼组(F组).麻醉诱导:靶控输注异丙酚,血浆靶浓度3μg/ml,三组分别靶控输注舒芬太尼(血浆靶浓度为0.5 ng/ml)、瑞芬太尼(血浆靶浓度为5 ng/ml)或芬太尼(血浆靶浓度为5 ng/ml),待BIS＜60时,静脉注射维库溴铵0.1 mg/kg,3 min后行双腔支气管插管,机械通气.麻醉维持:舒芬太尼、瑞芬太尼或芬太尼血浆靶浓度维持不变,停止输注异丙酚,持续吸入七氟醚0.7～1.5 MAC,按需间断静脉注射维库溴铵0.04 mg/kg.分别于麻醉诱导前(基础状态)、切皮后60min(T1)、术毕(T2)、术后24 h(T3)、72 h(T4)时采集肘静脉血样,检测CD3+、CD4+、CD8+及CD3- CD16+CD56+的百分率,计算CD4/CD8+,并检测血清白细胞介素-2(IL-2)和IL-10浓度.结果 与基础值比较,T2时三组CD4+百分率和CD4+/CD8+降低,CD3- CD16+CD56+百分率升高,SF组和RF组CD3+百分率降低,F组CD8+百分率升高,T3时三组CD3+和CIM+的百分率降低,RF组和F组血清IL-2浓度降低,T4时F组CD3+百分率和血清IL-2浓度降低,RF组和F组血清IL-10浓度升高(P＜0.05或0.01);与RF组比较,F组CD3+百分率升高,SF组血清IL-2浓度升高,血清IL-10浓度降低(P＜0.05).结论 舒芬太尼、瑞芬太尼或芬太尼均可抑制食管癌根治术病人的细胞免疫功能.     

Sevoflurane mimics ischemic preconditioning effects on coronary flow and nitric oxide release in isolated hearts.

BACKGROUND: Like ischemic preconditioning, certain volatile anesthetics have been shown to reduce the magnitude of ischemia/ reperfusion injury via activation of K+ adenosine triphosphate (ATP)-sensitive (K(ATP)) channels. The purpose of this study was (1) to determine if ischemic preconditioning (IPC) and sevoflurane preconditioning (SPC) increase nitric oxide release and improve coronary vascular function, as well as mechanical and electrical function, if given for only brief intervals before global ischemia of isolated hearts; and (2) to determine if K(ATP) channel antagonism by glibenclamide (GLB) blunts the cardioprotective effects of IPC and SPC. METHODS: Guinea pig hearts were isolated and perfused with Krebs-Ringer's solution at 55 mm Hg and randomly assigned to one of seven groups: (1) two 2-min total coronary occlusions (preconditioning, IPC) interspersed with 5 min of normal perfusion; (2) two 2-min occlusions interspersed with 5 min of perfusion while perfusing with GLB (IPC+GLB); (3) SPC (3.5%) for two 2-min periods; (4) SPC+GLB for two 2-min periods; (5) no treatment before ischemia (control [CON]); (6) CON+GLB; and (7) no ischemia (time control). Six minutes after ending IPC or SPC, hearts of ischemic groups were subjected to 30 min of global ischemia and 75 min of reperfusion. Left-ventricular pressure, coronary flow, and effluent NO concentration ([NO]) were measured. Flow and NO responses to bradykinin, and nitroprusside were tested 20-30 min before ischemia or drug treatment and 30-40 min after reperfusion. RESULTS: After ischemia, compared with before (percentage change), left-ventricular pressure and coronary flow, respectively, recovered to a greater extent (P<0.05) after IPC (42%, 77%), and treatment with SPC (45%, 76%) than after CON (30%, 65%), IPC+GLB (24%, 64%), SPC+GLB (20%, 65%), and CON+GLB (28%, 64%). Bradykinin and nitroprusside increased [NO] by 30+/-5 (means +/- SEM) and 29+/-4 nM, respectively, averaged for all groups before ischemia. [NO] increased by 26+/-6 and 27+/-7 nM, respectively, in SPC and IPC groups after ischemia, compared with an average [NO] increase of 8+/-5 nM (P<0.01) after ischemia in CON and each of the three GLB groups. Flow increases to bradykinin and nitroprusside were also greater after SPC and IPC. CONCLUSIONS: Preconditioning with sevoflurane, like IPC, improves not only postischemic contractility, but also basal flow, bradykinin and nitroprusside-induced increases in flow, and effluent [NO] in isolated hearts. The protective effects of both SPC and IPC are reversed by K(ATP) channel antagonism.     

Interleukin-8 serum levels in patients with various types of open heart surgical procedures performed under extracorporeal circulation

Serum concentrations of proinflammatory cytokine interleukin-8 (IL-8) in 15 patients with surgically revascularized myocardium by triple venous cardiopulmonary bypass (CPB x 3) and in 10 patients with implanted artificial aortic valves (AV) were measured. Average IL-8 concentrations in (CPB x 3) patients and in those with artificial aortic valve were 7.3 +/- 11.6 pg/mL 24 hours before surgery, i.e. 3.3 +/- 3.4 pg/mL; six hours after surgical procedure 32.7 +/- 71.4 pg/mL, i.e. 8.9 +/- 9.9 pg/mL; and 24 hours after surgery 10.9 +/- 9.7 pg/mL, i.e. 8.3 +/- 4.6 pg/mL. Extracorporeal circulation (ECC) caused significant increase of IL-8 serum concentration in both investigated groups six hours after surgery. Comparing preoperative values of the both groups, as well as those of 6 and 24 hours after surgery, no significant values of IL-8 were found. Various types of open heart surgical procedures had no influence on the extent of the production and secretion of proinflammatory IL-8 cytokine measured in patients during 24 hours after surgery.     

Coagulation abnormalities associated with severe isolated traumatic brain injury: cerebral arterio-venous differences in coagulation and inflammatory markers

Although coagulopathy is known to be the major contributor to a poor outcome of traumatic brain injury (TBI), the mechanisms that trigger coagulation abnormalities have not been studied in detail. We undertook a prospective observational study at a neurosurgical ICU (NICU) in a university hospital. We examined 11 patients with severe isolated TBI, at admittance to the hospital and during the next 3 days. We collected cerebrovenous blood samples from a jugular bulb catheter, arterial blood, and cerebrospinal fluid (CSF) samples. We measured concentrations of thrombin-antithrombin complex (TAT), fibrin D-dimer (DD), prothrombin fragment 1 + 2 (F1 + 2), interleukin-6 (IL-6), and complement complex (C5b-9). All patients had some degree of consumption coagulopathy at the study start and a tendency to thrombocytopenia during the next few days. Levels of DD (3.6 +/- 2.7 mg/L), TAT (86 +/- 72 microg/L) and F1 + 2 (5.9 +/- 6.8 nmol/L) were significantly increased shortly after the trauma compared to reference values, with considerable transcranial gradients for TAT (49 microg/L) and F1 + 2 (3.2 nmol/L). Compared to controls, IL-6 levels were increased more than a hundredfold in both blood (283 +/- 192 ng/L) and CSF (424 +/- 355 ng/L) samples, with a transcranial gradient at the study start (107 ng/L). C5b-9 levels were moderately increased in blood samples, 270 +/- 114 microg/L, versus controls, 184 +/- 39 (p < 0.05). We conclude that activation of the coagulation system takes place during the passage of blood through the damaged brain, and is already evident hours after the trauma. IL-6 and activation of the complement system (C5b-9) co-vary with hemostatic parameters in TBI patients.     

一氧化氮在门静脉高压症发病中的作用

目的　探讨一氧化氮在门静脉高压症发病中的作用。　方法　 75例门静脉高压症患者 ,术中胃网膜静脉插管测定门静脉压力 ,检测外周动静脉和门静脉血中内毒素和NO2 -/NO3-的含量。　结果　 ( 1)门静脉高压症患者的血中内毒素和NO2 -/NO3-的水平 [( 0 2 4 9± 0 112 )Eu/ml和( 5 5 9± 2 6 2 ) μmol/L]均显著高于对照组 ,且门静脉血中水平最高。 ( 2 )门静脉高压症患者门静脉压力[( 3 5 5± 4 4 )cmH2 O]与门静脉血NO2 -/NO3-的水平呈显著正相关 (n =2 5 ,r =0 5 5 ,P <0 0 1) ,二者在术后的变化量也呈正相关 (r =0 5 7,P <0 0 5 )。 ( 3 )门静脉高压症患者白蛋白水平与NO2 -/NO3-呈负相关 (n =75 ,r=- 0 3 5 ,P <0 0 1) ,且有腹水组的NO2 -/NO3-水平 [( 72 4± 2 0 3 ) μmol/L]较无腹水组 [( 5 0 3± 2 1 0 ) μmol/L]为高。　 结论　门静脉高压症患者血中内毒素和NO的水平升高 ,后者可能参与了门静脉压力的异常升高且与肝功能损害有关。     

Effect of acute ketosis on the endothelial function of type 1 diabetic patients: the role of nitric oxide.

In type 1 diabetic patients, acute loss of metabolic control is associated with increased blood flow, which is believed to favor the development of long-term complications. The mechanisms for inappropriate vasodilation are partially understood, but a role of endothelium-derived nitric oxide (NO) production can be postulated. We assessed, in type 1 diabetic patients, the effect of the acute loss of metabolic control and its restoration on forearm endothelial function in 13 type 1 diabetic patients who were studied under conditions of mild ketosis on two different occasions. In study 1, after basal determination, a rapid amelioration of the metabolic picture was obtained by insulin infusion. In study 2, seven type 1 diabetic patients underwent the same experimental procedure, except that fasting plasma glucose was maintained constant throughout. Basal plasma venous concentrations of nitrites/nitrates (NO2- + NO3-) were determined both before and after intravenous insulin infusion. Endothelium-dependent and -independent vasodilation of the brachial artery was assessed by an intra-arterial infusion of N(G)-monomethyl-L-arginine (L-NMMA) and sodium nitroprusside (SNP), respectively. The same parameters were determined in 13 control subjects at baseline conditions and during a hyperinsulinemic-euglycemic glucose clamp. Baseline forearm blood flow (4.89 +/- 0.86 vs. 3.65 +/- 0.59 ml x (100 ml tissue)(-1) x min(-1)) and NO2- + NO3- concentration (30 +/- 8 vs. 24 +/- 3 micromol/l) were higher in type 1 diabetic patients than in control subjects (P < 0.05). Insulin infusion was associated with lower forearm blood flow and plasma (NO2- + NO3-) concentration (P < 0.05), irrespective of the prevailing glucose levels, as compared with patients under ketotic conditions. The responses to L-NMMA were significantly lower in type 1 diabetic patients during euglycemia and hyperglycemic hyperinsulinemia (-11 +/- 5 and -10 +/- 4%, respectively, of the ratio of the infused arm to the control arm) than in control subjects at baseline (-18 +/- 6%, P < 0.05) and during hyperinsulinemia (-32 +/- 11%, P < 0.01). We conclude that the acute loss of metabolic control is associated with a functional disturbance of the endothelial function characterized by hyperemia and increased NO release during ketosis and blunted NO-mediated vasodilatory response during restoration of metabolic control by intravenous insulin. This functional alteration is unlikely to be explained by hyperglycemia itself.     

Thiazide-induced hypocalciuria is accompanied by a decreased expression of Ca2+ transport proteins in kidney

INTRODUCTION: Thiazide diuretics have the unique characteristic of increasing renal Na+ excretion, while decreasing Ca2+ excretion. However, the molecular mechanism responsible for this thiazide-induced hypocalciuria remains unclear. The present study investigates the effect of thiazides on the expression of the proteins involved in active Ca2+ transport as well as the role of extracellular volume (ECV) status. METHODS: Hydrochlorothiazide (HCTZ), 12 mg/24 hours, was administered during 7 days to Wistar rats by osmotic minipumps. In addition, ECV contraction was either prevented by Na+ repletion or induced by a low-salt diet. Expression levels of the proteins involved in active Ca2+ transport [i.e., epithelial Ca2+ channel (TRPV5/ECaC1), calbindin-D28K, Na+/Ca2+ exchanger (NCX1)], as well as the thiazide-sensitive Na+ Cl- cotransporter (NCC) were determined by real-time quantitative polymerase chain reaction (PCR) and semiquantitative immunohistochemistry. RESULTS: HCTZ significantly reduced urinary Ca2+ excretion (22%+/- 5% relative to controls). Hematocrit was significantly increased, confirming ECV contraction. In addition, Na+ depletion virtually abolished Ca2+ excretion (8%+/- 1%), while Na+ repletion during HCTZ treatment prevented both ECV contraction and hypocalciuria. HCTZ significantly decreased mRNA expression of TRPV5 (71%+/- 6%), calbindin-D28K (53%+/- 6%), NCX1 (51%+/- 8%) and NCC (50%+/- 11%), regardless of ECV status or calciuresis. Immunohistochemistry revealed reduced TRPV5 (43%+/- 2%), calbindin-D28K (59%+/- 1%) and NCC (56%+/- 4%) abundance. Furthermore, during HCTZ treatment, the subset of tubules coexpressing NCC and calbindin-D28K was significantly reduced (43%+/- 5%) and a disturbed cellular localization of NCC was observed. CONCLUSION: These data suggest that ECV contraction is a critical determinant of the thiazide-induced hypocalciuria, which is accompanied by a decreased expression of Ca2+ transport proteins.     

Autocrine effects of nitric oxide on HCO(3)(-) transport by rat thick ascending limb

BACKGROUND: In vivo and in vitro studies have shown that nitric oxide (NO) is an important modulator of transport processes along the nephron. The thick ascending limb (TAL) plays a significant role in the urine-concentrating mechanism and in the maintenance of acid/base balance. METHODS: TALs from male Sprague-Dawley rats were isolated and perfused, and net bicarbonate flux (J(HCO3)(-) was determined. RESULTS: In perfused TALs, 0.5 mmol/L L-arginine (L-Arg), the substrate for NO synthase, significantly lowered J(HCO3)(-) from 35.4 +/- 4.6 to 23.2 +/- 2.9 pmol. mm(-1). min(-1), a decrease of 36.9 +/- 11.6% (P < 0.025). D-Arg (0.5 mmol/L) had no effect on J(HCO3)(-) (N = 7). In the presence of 5 mmol/L L-NAME, an NO synthase (NOS) inhibitor, the addition of L-Arg did not affect TAL J(HCO3)(-) (43.4 +/- 4.4 vs. 44.6 +/- 5.0 pmol. mm(-1). min(-1)). L-NAME alone (5 mmol/L) did not affect TAL J(HCO3)(-). After removing L-Arg from the bath, J(HCO3)(-) increased from 26.2 +/- 3.9 to 34.8 +/- 3.2 pmol. mm(-1). min(-1) (P < 0.01), indicating no cytotoxicity of NO. We next investigated the effect of cGMP analogues on TAL J(HCO3)(-). 8-Br-cGMP (50 micromol/L) and db-cGMP (50 micromol/L) significantly decreased J(HCO3)(-) by 26.3 +/- 9.1% and 35.1 +/- 11.6%, respectively. In the presence of cGMP (50 micromol/L), the addition of L-Arg had no effect on J(HCO3)(-). In the presence of KT-5823 (2 mircromol/L), a protein kinase G inhibitor, the addition of L-Arg did not change TAL J(HCO3)(-) (N = 5). CONCLUSIONS: We conclude that (1) endogenously produced NO inhibits TAL J(HCO3)(-) in an autocrine manner, (2) cGMP mediates all the effects of NO, and (3) this effect is mediated by protein kinase G activation.     

谷氨酸受体拮抗剂在严重烧伤大鼠脑组织中的作用

目的 探讨烧伤后谷氨酸对脑组织的影响及谷氨酸受体拮抗剂的作用。方法 采用30％TBSAⅢ度烧伤大鼠模型,伤后2、6、12和24h测定了脑水含量、脑组织K^ 、Na^ 、Ca^2 、Mg^2 和-氧化氮(NO)代谢产物NO2^-／NO3^-浓度,在光镜和电镜下进行形态学和组织化学研究。结果 烧伤后脑水含量升高,脑组织Na^ 、Ca2^ 、NO2^-／NO3^-浓度高于对照组;脑内ATP酶减少;电镜观察毛细血管内皮细胞、神经细胞及胞浆部分线粒体肿胀,毛细血管壁出现胞饮泡。给予谷氨酸受体拮抗剂D-2-氨基-7-磷庚酸脂(D—AP7),可使脑水含量、脑组织Na^ 、Ca^2 、NO2^-／NO3^-浓度降低,毛细血管壁胞饮泡减少,神经细胞肿胀减轻。结论 烧伤后脑微血管通透性增加、组织细胞有缺血性改变和水肿发生。烧伤后脑组织形态及代谢变化与谷氨酸过量释放有关。兴奋毒性作用通过其受体介导,谷氨酸受体拮抗剂D—AP7可减轻烧伤后脑损伤。     

Interleukin-4 ameliorates crescentic glomerulonephritis in Wistar Kyoto rats

BACKGROUND: Activated macrophages play a central role in crescentic glomerulonephritis. Interleukin-4 (IL-4) down-regulates many macrophage proinflammatory activities. We therefore studied the effect of IL-4 on glomerular injury in a model of crescentic glomerulonephritis in the Wistar Kyoto rat. METHODS: Glomerulonephritis was induced by i.v. administration of rabbit antirat glomerular basement membrane antiserum (nephrotoxic serum, NTS). In experiment 1, IL-4 was given from two hours before NTS until day 6. In experiment 2, rats were treated from day 0 to 7 and were then monitored until killed on day 28. In experiment 3, IL-4 was given from day 4 to 7. RESULTS: Continuous IL-4 treatment (experiment 1) significantly (P = 0.001) reduced proteinuria (3 +/- 1 mg per 24 hr vs. 56 +/- 7), fibrinoid necrosis (0.06 +/- 0.04 quadrants/glomulus vs. 1.2 +/- 0.1), macrophage infiltration (6.7 +/- 2.6 cells/glom vs. 33 +/- 2.5), CD8+ cells (1.5 +/- 0.6 cells/glom vs. 6.2 +/- 1.1), inducible nitric oxide synthase positive cells (0.04 +/- 0.04 cells/glom vs. 3.7 +/- 0.6), proliferating cell nuclear antigen positive cells (3.2 +/- 1 cells/glom vs. 15 +/- 2.3), and glomerular intercellular adhesion molecule-1 expression. Follow-up after seven days of treatment (experiment 2) showed that at four weeks, creatinine clearance was higher in treated rats (1.1 +/- 0.1 ml/min vs. 0.4 +/- 01, P = 0.011), and both glomerular scarring (P = 0.006) and tubular atrophy (P = 0.006) were less. Delayed treatment (experiment 3) reduced proteinuria (41 +/- 5 mg per 24 hr vs. 97 +/- 9, P = 0.004) and fibrinoid necrosis (0.39 +/- 0.05 quadrants/glom vs. 1.6 +/- 0.1, P = 0.004). There was no difference in macrophage infiltration, but inducible nitric oxide synthase positive cells were reduced (0.6 +/- 0.1 cells/glom vs. 1.8 +/- 0.4, P = 0.01) as were ED3+ cells (0.18 +/- 0.06 cells/glom vs. 1.86 +/- 0.21, P = 0.004). CONCLUSION: In this model of crescentic glomerulonephritis, early IL-4 treatment abolished proteinuria and markedly reduced glomerular inflammation. If treatment was stopped after seven days, there was continuing benefit on glomerular and tubulointerstitial scarring and creatinine clearance at four weeks. If treatment was delayed until inflammation was established, there was still a reduction of injury, but without an alteration of macrophage numbers, suggesting that IL-4 may be acting, in part, to reduce macrophage activation.     

Modulation of IL-1 effects on cartilage by NO synthase inhibitors: pharmacological studies in rats.

Objective To compare the ability of L-arginine (L-arg) analog nitric oxide synthase (NOS) inhibitors and isothioureas to restore the interleukin-1 (IL-1) induced inhibition of proteoglycan (PG) synthesis in rat.Methods Chondrocytes beads and patellae were challenged with IL-1betain vitro and monitored for NO production and proteoglycan synthesis. Rats injected with IL-1beta in knee joints were monitored for NO(2)( - )+NO(3)( - )levels in joint tissues and ex-vivo(35)S sulfate incorporation in patellae. NOS inhibitors were either added to culture medium or injected concomittantly to IL-1beta.Results Ability of NOS inhibitors to reduce NO(2)( - )levels decreased from chondrocytes beads to patellae. Partial restoration of PG synthesis was restricted to L-arg analogs in patellae. After IL-1 injection, NO was produced from patella and synovium. L-arg analogs restored partly PG synthesis when decreasing significantly NO(2)( - )+NO(3)( - )levels in synovial fluid. Isothioureas were ineffective.Conclusions NO accounts importantly for IL-1 induced inhibition of cartilage anabolism in rat. L-arg analog NOS inhibitors are more effective than isothioureas in restoring PG synthesis and have chondroprotective potency when administered locally in diseased joint.     

Stimulated response of peripheral lymphocytes may distinguish cyclosporine effect in renal transplant recipients receiving a cyclosporine+rapamycin regimen

BACKGROUND: Clinically, cyclosporine (CSA, Neoral) is titrated to concentrations, and not to pharmacological effect. METHODS: Intracellular interleukin- (IL) 2 was measured in phorbol myristic acid-ionomycin-stimulated peripheral lymphocytes by flow cytometry, after isolation from 14 renal transplant recipients receiving CSA+prednisone, and double-blind rapamycin (rapamycin:placebo=4:1). RESULTS: The proportion (%) of CD4+IL-2+ lymphocytes corresponding to CSA levels (mean+/-SD ng/ml) measured preoperatively (TO=O), and on postoperative day 8, before (356+/-63), and 2 hr after the morning dose (Cmax=1567+/-669), decreased from 39+/-16 to 15+/-8 and 3+/-1.6, respectively. Reciprocally, unresponsive lymphocytes (%CD4+IL-2-) increased with increasing CSA levels and predicted an EC50 of 249 ng/ml (CSA concentration at which CD4+IL-2- cells increased by 50% over baseline) in an Emax pharmacodynamic model. CONCLUSIONS: Clinically, the pharmacological effect of CSA is quantifiable, and lies in the upper end of the predicted range. In our Neoral-treated sample population, Cmax was associated with the least variable "cyclosporine effect." Such information could potentially individualize immunosuppression, and lead to rational dosing strategies.     

Sevoflurane Mimics Ischemic Preconditioning Effects on Coronary Flow and Nitric Oxide Release in Isolated Hearts

Background: Like ischemic preconditioning, certain volatile anesthetics have been shown to reduce the magnitude of ischemia/ reperfusion injury viaactivation of K+ adenosine triphosphate (ATP)-sensitive (KATP) channels. The purpose of this study was (1) to determine if ischemic preconditioning (IPC) and sevoflurane preconditioning (SPC) increase nitric oxide release and improve coronary vascular function, as well as mechanical and electrical function, if given for only brief intervals before global ischemia of isolated hearts; and (2) to determine if KATP channel antagonism by glibenclamide (GLB) blunts the cardioprotective effects of IPC and SPC.

Methods: Guinea pig hearts were isolated and perfused with Krebs-Ringer's solution at 55 mmHg and randomly assigned to one of seven groups: (1) two 2-min total coronary occlusions (preconditioning, IPC) interspersed with 5 min of normal perfusion; (2) two 2-min occlusions interspersed with 5 min of perfusion while perfusing with GLB (IPC+GLB); (3) SPC (3.5%) for two 2-min periods; (4) SPC+GLB for two 2-min periods; (5) no treatment before ischemia (control [CON]); (6) CON+GLB; and (7) no ischemia (time control). Six minutes after ending IPC or SPC, hearts of ischemic groups were subjected to 30 min of global ischemia and 75 min of reperfusion. Left-ventricular pressure, coronary flow, and effluent NO concentration ([NO]) were measured. Flow and NO responses to bradykinin, and nitroprusside were tested 20-30 min before ischemia or drug treatment and 30-40 min after reperfusion.

Results: After ischemia, compared with before (percentage change), left-ventricular pressure and coronary flow, respectively, recovered to a greater extent (P < 0.05) after IPC (42%, 77%), and treatment with SPC (45%, 76%) than after CON (30%, 65%), IPC+GLB (24%, 64%), SPC+GLB (20%, 65%), and CON+GLB (28%, 64%). Bradykinin and nitroprusside increased [NO] by 30 +/- 5 (means +/- SEM) and 29 +/-4 nM, respectively, averaged for all groups before ischemia. [NO] increased by 26 +/- 6 and 27 +/- 7 nM, respectively, in SPC and IPC groups after ischemia, compared with an average [NO] increase of 8 +/- 5 nM (P < 0.01) after ischemia in CON and each of the three GLB groups. Flow increases to bradykinin and nitroprusside were also greater after SPC and IPC.     

Catecholamines decrease nitric oxide production by cytokine-stimulated hepatocytes

BACKGROUND: Catecholamines are significantly elevated in inflammatory responses and play a regulatory role in sepsis. Nitric oxide (NO), also a key inflammatory mediator in sepsis, is produced in large amounts by the inducible nitric oxide synthase (iNOS) in the liver. The purpose of this study was to test the hypothesis that catecholamines play a role in the regulation of NO production by hepatocytes. METHODS: Primary hepatocytes were isolated from healthy male Sprague-Dawley rats and either cultured with normal medium or stimulated with cytomix (interleukin-1 beta, interferon-gamma, and tumor necrosis factor-alpha) in the presence or absence of epinephrine or norepinephrine at varying concentrations. Total RNA was isolated 6 hours after treatment and analyzed by Northern blotting for iNOS mRNA. Protein extracts were obtained at 12 hours and were analyzed by Western immunoblotting for iNOS. Cell culture supernatants were analyzed for NO, determined as the stable end-product NO(2)(-), at 24 hours. RESULTS: Epinephrine and norepinephrine significantly decreased NO(2)(-) levels in stimulated hepatocytes but had no effect on iNOS mRNA or protein levels. The decrease in NO(2)(-) was reproduced by the adenylate cyclase stimulator, forskolin. The catecholamine-induced decrease in NO(2)(-) was completely reversed by the protein kinase A inhibitor Rp-8-Br-cyclic adenosine monophosphate. CONCLUSIONS: Catecholamines decrease hepatocyte production of NO in response to cytokine stimulation. This effect seems to be due to post-translational events and appears to be mediated in part by cyclic adenosine monophosphate.     

Analysis of interleukin-8, interleukin-10, and tumor necrosis factor-alpha in the cerebrospinal fluid of patients with cervical spondylotic myelopathy

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: The expression of interleukin-8 (IL-8), IL-10, and tumor necrosis factor-alpha (TNF-alpha) were measured in the cerebrospinal fluid (CSF) of patients with cervical myelopathy. The purpose of this study was to examine whether the CSF levels of those 3 cytokines differ significantly among 3 groups of patients with different diseases. METHODS: IL-8, IL-10, and TNF-alpha levels were analyzed using enzyme-linked immune assay. CSF samples were collected from 3 groups of patients. The cervical spondylotic myelopathy (CSM) group consisted of 35 patients. The ossification of the posterior longitudinal ligament group [(OPLL) group] consisted of 7 patients, and the control group consisted of 12 patients. The concentration of IL-8 was 69.0+/-35.2 pg/mL in the CSM group, 82.1+/-46.7 pg/mL in the OPLL group, and 43.5+/-20.9 pg/mL in the control group. The concentration of IL-8 was significantly higher in the CSM and OPLL groups than in the control group (P<0.05). There was no significant difference between the CSM group and OPLL group. The concentration of IL-10 and TNF-alpha in all groups was below the sensitivity of the measurements. CONCLUSIONS: In this study, the concentration of IL-8 was high in CSM and OPLL patients. However, the concentration of IL-10 and TNF-alpha was below the sensitivity of the measurements.