Prevention and management of diabetic retinopathy in STZ diabetic rats by Tinospora cordifolia and its molecular mechanisms

We investigated the potential of Tinospora cordifolia (TC) in treatment of diabetic retinopathy in STZ-induced rats due to its antihyperglycemic, angiogenic, antiinflammatory and antioxidant effects. The diabetic rats, treated for 24 weeks with TC extract (250 mg/kg), were evaluated for lenticular and fundus changes. Biochemical parameters were estimated and histopathological studies performed. TC significantly reduced blood glucose and glycated hemoglobin in treated rats. It prevented cataract development in treated group. Angiogenic markers VEGF and PKC increased in diabetic retina, which reduced significantly with TC. Anti-inflammatory parameters TNF-α and IL-1β elevated in diabetic group unlike that in treated group. TC also provided defense against depletion of antioxidant enzymes- glutathione and catalase. Histopathological studies revealed thickening of basement membrane of the retinal and glomerular vasculature of diabetic rat, but no basement membrane widening was seen in treated animals. Destruction of pancreatic islet structure was observed in diabetic group, but not in treated. Thus, TC reduces blood glucose and inhibits overexpression of angiogenic and inflammatory mediators, which are distinct markers of diabetic retinopathy. It also prevents retinal oxidative stress and restores antioxidant enzyme levels. These data provide evidence for the safety and potential effect of TC in the management of experimental diabetic retinopathy.     

A preliminary evaluation of a novel method to monitor a triple antioxidant combination (vitamins E, C and α-lipoic acid) in diabetic volunteers using in vitro methaemoglobin formation

Eight otherwise healthy diabetic volunteers took a daily antioxidant supplement consisting of vitamin E (200 IU), vitamin C (250 mg) and α-lipoic acid (90 mg) for a period of 6 weeks. Diabetic dapsone hydroxylamine-mediated methaemoglobin formation and resistance to erythrocytic thiol depletion was compared with age and sex-matched non-diabetic subjects. At time zero, methaemoglobin formation in the non-diabetic subjects was greater at all four time points compared with that of the diabetic subjects. Resistance to glutathione depletion was initially greater in non-diabetic compared with diabetic samples. Half-way through the study (3 weeks), there were no differences between the two groups in methaemoglobin formation and thiol depletion in the diabetic samples was now lower than the non-diabetic samples at 10 and 20 min. At 6 weeks, diabetic erythrocytic thiol levels remained greater than those of non-diabetics. HbA(1c) values were significantly reduced in the diabetic subjects at 6 weeks compared with time zero values. At 10 weeks, 4 weeks after the end of supplementation, the diabetic HbA1(c) values significantly increased to the point where they were not significantly different from the time zero values. Total antioxidant status measurement (TAS) indicated that diabetic plasma antioxidant capacity was significantly improved during antioxidant supplementation. Conversion of α-lipoic acid to dihydrolipoic acid (DHLA) in vivo led to potent interference in a standard fructosamine assay kit, negating its use in this study. This report suggests that triple antioxidant therapy in diabetic volunteers attenuates the in vitro experimental oxidative stress of methaemoglobin formation and reduces haemoglobin glycation in vivo.     

Green tea polyphenols change the profile of inflammatory cytokine release from lymphocytes of obese and lean rats and protect against oxidative damage

This study aimed to investigate whether green tea polyphenols (GT) modulate some functional parameters of lymphocytes from obese rats. Male Wistar rats were treated with GT by gavage (12 weeks/5 days/week; 500 mg/kg of body weight) and obesity was induced by cafeteria diet (8 weeks). Lymphocytes were obtained from mesenteric lymph nodes for analyses. In response to the cafeteria diet we observed an increase in activity of the metabolic enzyme hexokinase, ROS production, MnSOD, CuZnSOD and GR enzyme activities and proliferation capacity of the cells (baseline), whereas IL-10 production was decreased. Obese rats treated with GT decreased cell proliferation (under ConA stimulation). Hexokinase and G6PDH activity, ROS production and MnSOD, CuZnSOD, GPx and GR enzymes remained increased, accompanied by an increase in Nrf2 mRNA level. There was a decrease in pro-inflammatory IL-2, IL-6, IL-1β, TNF-α cytokines that were accompanied by a decrease in the mRNA level of TRL4 while IL-10 production was increased in obese rats treated with GT. GT treatment of lean rats showed similar results to that of obese rats treated with GT, indicating that the effects of GT are independent of diet. Foxp3 and IRF4 mRNA levels were increased by GT. In conclusion, cafeteria diet modulated the function of lymphocytes from lymph nodes, increasing ROS production and decreasing anti-inflammatory IL-10, which could contribute to the inflammatory state in obesity. GT reduced ROS production, improving the redox status and reducing pro-inflammatory cytokine production by lymphocytes, suggesting that GT treatment may be driving lymphocytes to a more anti-inflammatory than pro-inflammatory microenvironment.     

舒洛地特联合瑞格列奈治疗糖尿病肾病的临床研究

目的研究舒洛地特联合瑞格列奈治疗糖尿病肾病的临床疗效。方法选取2014年5月—2015年2月重庆市南岸区人民医院收治的糖尿病肾病患者200例,随机分为对照组和治疗组,每组各100例。对照组口服瑞格列奈片,0.5 mg/次,2次/d,连续使用2周后根据患者表现出来的具体症状调整用药剂量。治疗组在对照组治疗基础上口服舒洛地特软胶囊,1粒/次,2次/d。两组均连续治疗4个月。观察两组的临床疗效,同时比较治疗前后两组患者空腹血糖(FPG)、餐后2 h血糖(2 h PG)、糖化血红蛋白(Hb A1c)、体质量指数(BMI)、24 h尿蛋白、血肌酐(Scr)、尿素氮(BUN)、IL-6、IL-10、TNF-α的变化情况。结果治疗后,对照组和治疗组的总有效率分别为88.0%、96.0%,两组比较差异有统计学意义(P0.05)。治疗后,两组患者FPG、2 h PBG、Hb A1c、24 h尿蛋白、Scr、BUN、IL-6、IL-10、TNF-α均显著降低,同组治疗前后差异有统计学意义(P0.05);治疗后治疗组这些观察指标的改善程度优于对照组,两组比较差异有统计学意义(P0.05);治疗后治疗组患者BMI较治疗前显著降低,对照组无明显变化,两组比较差异有统计学意义(P0.05)。结论舒洛地特联合瑞格列奈治疗糖尿病肾病具有较好的临床疗效,可较好的控制患者的血糖水平,减轻肾脏损害,具有一定的临床推广应用价值。     

化学性低氧模拟剂氯化钴诱导人角质形成细胞炎症反应的研究

目的探讨化学性低氧模拟剂氯化钴对人皮肤角质形成细胞(HaCat)炎症反应的影响。方法用不同浓度的CoCl2处理HaCat细胞,建立化学性低氧诱导皮肤细胞损伤的实验模型后,检测细胞存活率、细胞内活性氧(ROS)、线粒体膜电位(MMP)、细胞培养液中白介素6(IL-6)和白介素8(IL-8)水平以及血红素加氧酶(HO-1)表达。结果在500～3000μmol.L-1浓度范围内,CoCl2可降低HaCat细胞存活率,且CoCl2剂量越大、细胞存活率降低越明显;2000μmol.L-1CoCl2能诱导HaCat细胞产生氧化应激反应,使胞内ROS生成增多,MMP降低;CoCl2能诱导HaCat细胞产生炎症反应,使IL-6和IL-8释放增多;1000～3000μmol.L-1CoCl2能上调HO-1的表达。结论CoCl2在诱导HaCat细胞产生氧化应激反应的同时,也能引起炎症反应,促进IL-6和IL-8的释放及HO-1表达上调。     

Cerebroprotective effect of resveratrol through antioxidant and anti-inflammatory effects in diabetic rats

Oxidative stress and inflammation have been implicated in cerebral ischemia/reperfusion injury and complication of diabetes. The present study was designed to evaluate whether resveratrol has cerebroprotective action through antioxidant and anti-inflammatory actions in diabetic rats. Bilateral common carotid artery occlusion (30 min) and reperfusion (4 h) was employed to induce cerebral infarction in diabetic Wistar rats. Diabetes was induced by streptozocine (50 mg/kg) intraperitoneally at once. Diabetic animals were divided into groups as: normal, sham, ischemia–reperfusion, and resveratrol-treated (5, 10, 20, and 30 mg/kg). These were used for estimation of cerebral infarction. Furthermore, 20 mg/kg dose was selected for estimation of oxidative stress markers (malondialdehyde, superoxide dismutase, and catalase). Inflammatory markers like TNF-α, IL-6, IL-10, and myeloperoxidase were estimated and histological characters were studied. Resveratrol produced dose-dependent reduction in percent cerebral infarction. With resveratrol of 20 mg/kg dose, levels of oxidative stress markers and inflammatory markers like malondialdehyde, TNF-α, IL-6, and myeloperoxidase were reduced and there was a significant increase in the levels of antioxidant and anti-inflammatory markers like catalase, superoxide dismutase, and IL-10. In the present study, we found that mechanism(s) responsible for the cerebroprotective effect of resveratrol in the diabetic rat brain involves antioxidant and anti-inflammatory actions.     

中药仙灵脾对2型糖尿病大鼠炎症因子的影响

目的:评价中药仙灵脾对2型糖尿病大鼠炎症因子的影响。方法:雄性Wistar大鼠70只,从中随机选取10只为正常对照组,其余应用高糖高脂饲料喂养及链脲佐菌素腹腔注射造模,成模2型糖尿病大鼠共44只按血糖值随机分组:糖尿病(DM)模型对照组(16只)、仙灵脾组(16只)、二甲双胍组(12只),各组共相应给药8周,治疗前后取血测量血糖(FBG)、C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)。结果:成模2型糖尿病各组大鼠FBG,CRP,TNF-α,IL-6水平均较正常对照组大鼠明显增高(P<0.01)。应用仙灵脾及二甲双胍治疗的两组大鼠的FBG,CRP,TNF-α,IL-6水平均较DM模型对照组显著下降(P<0.05),但两治疗组比较差别无统计学意义(P>0.05)。结论:中药仙灵脾治疗2型糖尿病大鼠可降低炎症因子水平,抑制体内炎症反应。     

Neutrophil migration during liver cirrhosis in rabbits

1. The aim of the present study was to investigate neutrophil chemotaxis during the induction of liver cirrhosis in rabbits. 2. Liver cirrhosis was induced in male New Zealand white rabbits. The study consisted of three experimental groups: (i) group A (n=16) served as the control and received only normal chow and all rabbits in this group were killed at 16 weeks; (ii) group B rabbits (n=8) were killed immediately after the chemotaxis assay, which was performed 24 h after CCl4 administration, at weeks 2, 4, 6 and 8; and (iii) in group C rabbits (n=19), the chemotaxis assay was performed every second week on the day before CCl4 administration for 16 weeks and all animals in this group were killed at 16 weeks. 3. Four of six rabbits in group B had liver cirrhosis at week 8. In group C, liver cirrhosis occurred in seven of eight animals. All rabbits with liver cirrhosis had an inflammatory infiltrate of neutrophils. In group B, there was a significant increase in polymorphonuclear cells and neutrophil chemotaxis and a significant reduction in mononuclear leucocytes at week 8. The rabbits in group C showed a significant increase in total leucocyte and polymorphonuclear numbers at week 10. A significant increase in neutrophil chemotaxis was also observed from week 2 through to week 6. 4. The presence of neutrophils in the liver of all rabbits with cirrhosis, associated with an increase in polymorphonuclear cell chemotaxis during this process, supports the view that this cell type has an important role in the development of toxic liver damage.     

Antioxidant vitamins reduce oxidative stress and ventricular remodeling in patients with acute myocardial infarction

We analyzed soluble vascular adhesion molecules (sVCAM-1), reactive oxygen metabolites (ROMs) level, total antioxidant status (TAS) and telediastolic left ventricular volume (TLVV) in patients with myocardial infarction undergoing reperfusion therapy and treated with antioxidant vitamins (AT) or placebo (P) before and for 1 month after reperfusion. After reperfusion, sVCAM-1 serum concentration, reactive oxygen metabolites level, and TLVV were significantly higher in patients treated with placebo than in those treated with antioxidant vitamins, while TAS was significantly higher in patients treated with antioxidant supplementation. We observed that 48 hours after reperfusion sVCAM-1 (P) vs sVCAM-1 (AT) was 2.03+/-0.5 vs 1.63+/-0.7 microg/ml with p < 0.01; ROMs (P) vs ROMs (AT) were 335.60+/-35.80 vs 307.50+/-47.10 U.CARR with p < 0.05; TAS (P) vs TAS (AT) was 526.47+/-44.24 vs 737.65+/-51.15 micromol/l with p < 0.01; 1 week after reperfusion TLVV (P) vs TLVV (AT) was 125.12+/-29.80 vs 119.40+/-29.40 ml with p < 0.05; 1 month after reperfusion TLVV (P) vs TLVV (AV) was 132.00+/-33.50 vs 123.40+/-21.60 ml with p < 0.05. In the first period after infarction, vitamin treatment improves the antioxidant system and reduces oxidative stress, inflammatory process and left ventricular remodeling.     

Protective role of ascorbic acid on subacute sperm toxicity in male New Zealand white rabbits treated with endosulfan

The ameliorating effect of oral ascorbic acid (AA) was evaluated against changes in sperm parameters in New Zealand White (NZW) rabbits treated with endosulfan. Rabbits (6 to 8 months old) were divided into four groups of six animals each. Rabbits in TRT-I served as control and received corn oil by oral gavage for 6 weeks. Rabbits in TRT-II received endosulfan (1 mg/kg bw per day) in corn oil. TRT-III group received oral corn oil daily and ascorbic acid (AA; 20 mg/kg bw) every other day for 6 weeks. TRT-IV group received the same amounts of endosulfan and AA. Endosulfan alone significantly reduced the sperm count and motility and increased the presence of sperm with morphologic problems. AA treatment showed significant amelioration when coupled with endosulfan. Ameliorations were up to control levels in all cases except for sperm motility. The data suggested that AA has beneficial influences in neutralizing the toxic effects of endosulfan in the spermatologic parameters of NZW males.     

Protective Role of Ascorbic Acid on Subacute Sperm Toxicity in Male New Zealand White Rabbits Treated with Endosulfan

The ameliorating effect of oral ascorbic acid (AA) was evaluated against changes in sperm parameters in New Zealand White (NZW) rabbits treated with endosulfan. Rabbits (6 to 8 months old) were divided into four groups of six animals each. Rabbits in TRT-I served as control and received corn oil by oral gavage for 6 weeks. Rabbits in TRT-II received endosulfan (1 mg/kg bw per day) in corn oil. TRT-III group received oral corn oil daily and ascorbic acid (AA; 20 mg/kg bw) every other day for 6 weeks. TRT-IV group received the same amounts of endosulfan and AA. Endosulfan alone significantly reduced the sperm count and motility and increased the presence of sperm with morphologic problems. AA treatment showed significant amelioration when coupled with endosulfan. Ameliorations were up to control levels in all cases except for sperm motility. The data suggested that AA has beneficial influences in neutralizing the toxic effects of endosulfan in the spermatologic parameters of NZW males.     

Differential effects of ET(A) and ET(B) receptor antagonism on oxidative stress in type 2 diabetes

Endothelin (ET-1) is chronically elevated in diabetes. However, role of ET-1 in increased oxidative stress in type 2 diabetes is less clear. This study tested the hypotheses that: 1) oxidative stress markers are increased and total antioxidant capacity is decreased in diabetes, and 2) activation of ET(A) receptors mediates oxidative stress whereas ET(B) receptors display opposing effects. Plasma total antioxidant status (TAS) and 8-isoprostane (8-iso PGF(2alpha)) as well as total nitrotyrosine levels in mesenteric resistance vessels were measured in control Wistar and diabetic Goto-Kakizaki (GK) rats (n=5-10) treated with vehicle, ET(A) antagonist (atrasentan, 5 mg/kg/day), or ET(B) receptor antagonist (A-192621, 15 or 30 mg/kg/day, low and high dose, respectively) for 4 weeks. 8-iso PGF(2alpha) (pg/ml) levels were significantly higher in low dose A-192621 treatment groups of control and diabetic rats than in atrasentan or high-dose A-192621 treated groups. Protein nitration was increased in diabetes and ET(A) receptor antagonism prevented this increase. TAS levels were similar in all experimental groups. Thus, ET-1 contributes to oxidative stress in type 2 diabetes and ET receptor antagonism with atrasentan or A-192612 displays differential effects depending on dose and receptor subtype.     

The influence of sulindac on diabetic cardiomyopathy: a non-invasive evaluation by Doppler echocardiography in streptozotocin-induced diabetic rats

The aim of the present study was to investigate the cardioprotective activity of sulindac as an aldose reductase inhibitor in the development of cardiomyopathy by non-invasive techniques; M-mode and Doppler echocardiography. Diabetes was induced by streptozotocin (45 mg/kg, iv) in the Sprague-Dawley rats. Echocardiography, biochemical and histological studies were carried out in normal control, diabetic untreated, diabetic vehicle (sodium carboxy methyl cellulose, 1%, po) and sulindac (6 mg/kg and 20 mg/kg, po) treated animals at varying time intervals. In the diabetic untreated and vehicle treated rats at 12 weeks after induction of diabetes, there was a significant decrease in the E-wave, an increase in the A-wave and corresponding decrease in the E/A ratio was observed. Significant decrease in the Eat was found after 12 weeks (P < 0.05). Whereas systolic function variables; ejection fraction and fractional shortening were significantly decreased (P < 0.05) after 12 weeks compared to their baseline data. In the sulindac treated animals, there were no significant alterations in the systolic and diastolic parameters were found throughout the study period. Myocardial fructose levels were significantly increased in the diabetic untreated animals compared to normal control rats (P < 0.05), whereas these were significantly decreased in the sulindac (6 mg/kg and 20 mg/kg) treated animals (301.11+/-37.98, 214.11+/-25.31, vs. 914.88+/-56.01 nmol/g) compared to diabetic vehicle treated group (P < 0.05). Extensive focal ischemic myocyte degeneration was observed in the diabetic untreated and vehicle treated rats, whereas in the sulindac (6 mg/kg) treated rats, minimal necrosis was found, with no evidence of necrosis in sulindac (20 mg/kg) group. Our results show for the first time that sulindac has a cardioprotective activity as this agent prevented the development of left ventricular dysfunction in STZ-induced diabetic rats in the 12-week chronic study.     

益气活血汤联合尿激酶对结核性胸膜炎患者炎症因子、E-选择素、LDH水平的影响

目的 探讨益气活血汤联合尿激酶对结核性胸膜炎患者炎症因子、E-选择素、LDH水平影响.方法 70例胸膜炎患者随机分为研究组与对照组,每组35例.对照组患者给予尿激酶治疗,研究组在尿激酶治疗基础上结合益气活血汤治疗.两组患者疗程均为8周.对比分析两组患者治疗总有效率、胸腔积液吸收时间和住院时间,治疗前和治疗8周血清炎症因子、E-选择素、LDH水平及药物副反应情况.结果 研究组治疗总有效率明显高于对照组(91.43％ vs.65.71％),差异有统计学意义(P＜0.05);研究组胸腔积液吸收时间和住院时间均显著短于对照组(P＜0.05);两组患者治疗后IL-6和IL-8水平较治疗前下降,差异有统计学意义(P＜ 0.05);研究组患者治疗后IL-6和IL-8水平低于对照组(P＜0.05);两组患者治疗后E-选择素、LDH水平较治疗前下降(P＜0.05);研究组患者治疗后E-选择素、LDH水平低于对照组(P＜0.05);两组患者在治疗期间均未出现严重药物副反应.结论 益气活血汤联合尿激酶对结核性胸膜炎患者疗效明显,可降低炎症因子IL-6和IL-8、E-选择素、LDH,具有重要研究价值.     

诺和龙治疗2型糖尿病合并动脉粥样硬化患者的临床疗效分析

目的对诺和龙治疗2型糖尿病合并动脉粥样硬化患者的临床疗效进行分析。方法按照双盲随机对照实验原则设计本研究方法,将112例2型糖尿病患者分为2组,即观察组和对照组,每组各56例,2组患者均给予常规口服降糖药物或皮下注射胰岛素治疗,观察组患者在此基础上加用诺和龙治疗,比较两组患者的治疗效果。结果观察组患者治疗后的FBG、2hPG、HbA1c、IL-6以及内膜中层厚度较对照组患者有显著改善,组间差异有统计学意义(P<0.05)。结论采用诺和龙治疗2型糖尿病合并动脉粥样硬化临床疗效可靠,值得进一步推广使用。     

炎症因子对非 PCOS 患者颗粒细胞活性氧水平及线粒体 DNA 拷贝数的影响

目的 探讨炎症因子对非多囊卵巢综合征（PCOS） 患者颗粒细胞活性氧（ROS） 水平及线粒体 DNA （mtDNA）的影响。 方法 选择体外受精-胚胎移植（IVF-ET）治疗的非 PCOS 患者 50 例, 提取卵泡液中颗粒细胞进行体外培养, 随机分为炎症因子处理组和对照组, 处理组分别加入 5 nmol/L 的炎症因子白细胞介素（IL）-1、IL-6、肿瘤坏死因子（TNF）-α, 对照组加入等量炎症因子稀释液, 比较各组颗粒细胞 ROS 水平及 mtDNA 拷贝数是否存在差异。 结果 IL-1、IL-6、TNF-α 处理组颗粒细胞 ROS 水平及 mtDNA 拷贝数均显著高于对照组（ 均 P＜ 0.05）。 结论 炎症因子 IL-1、IL-6、TNF-α 可导致颗粒细胞氧化应激水平增高和线粒体受损。     

Effect of Syringic acid on antioxidant biomarkers and associated inflammatory markers in mice model of asthma

Asthma is termed as the induction of chronic inflammation in the airway lumen of lungs due to accumulation of inflammatory cells which affects normal breathing process. Prolonged accumulation of inflammatory cells leads to oxidative stress and suppression of antioxidant activities. Therefore, in our present investigation, a potential phenolic compound, Syringic acid was tested for the suppression of inflammatory markers toward an antiasthmatic activity in ovalbumin (OVA)-induced asthmatic mice model. As a result, the Syringic acid treatment was found to suppress the inflammatory cells; eosinophil, neutrophil, macrophage, lymphocyte, and other inflammatory markers including IL-4, IL-5, IL-13, and TNF-α in the BALF of OVA-induced asthmatic mice. Similarly, IgE levels were significantly reduced in the blood serum of Syringic acid treated mice groups. In this context, the IFN-γ levels were found enhanced in the BALF of Syringic acid treated asthmatic mice groups, expressing an anti-inflammatory response. Enzymatic and nonenzymatic antioxidants such as SOD, CAT, and GSH levels were found high in the Syringic acid treatment than the asthmatic control group, which depicts the antioxidant response of Syringic acid on asthmatic groups. Intriguingly, the ROS, NO₂, NO₃, and MDA levels were inhibited in the BALF of Syringic acid treated mice groups. The airway hyper-reactivity (AHR) was comparatively normal in the Syringic acid treatment as it was severe in the case of asthmatic control group. Consequently, the effect of Syringic acid is prominent in the treatment of asthma by controlling the accumulation of inflammatory cells, other inflammatory markers along with enhancement of antioxidant markers, suppression of ROS and controlling airway hyperreactivity. Hence, Syringic acid may be recommended for clinical trials in the treatment of asthma.     

兔肌肉种植VX_2瘤前后及发生转移后白细胞介素-2、6、8的变化

目的:观察兔肌肉种植VX2瘤前后及在肝、肺发生转移后白细胞介素-2、6、8(IL-2、6、8)的改变情况。方法:将健康雄性3年的新西兰大白兔80只,分成2组,外照射、体外高频热疗组(治疗组)、对照组,各40只。观察肿瘤直径,生存期同时检测IL-2、IL-6、IL-8在VX2瘤种植前后和发生转移后的变化情况。结果:治疗组肿瘤直径小于对照组(P<0.01);治疗组兔生存期长于对照组(P<0.01);肺、肝转移在治疗组中明显少于对照组(P<0.01);VX2瘤移植成功后细胞因子的变化为IL-2、IL-8降低,IL-6增高(P<0.01);在发生肺、肝转移兔中相比IL-2、IL-8降低,IL-6增高,与未发生肺、肝转移的兔相比差异有显著性(P<0.01)。结论:VX2种植成功后对实验兔的IL-2、6、8产生影响;发生肺、肝转移后IL-2、IL-8逐渐降低,IL-6增高。     

Importance of soluble metals and reactive oxygen species for cytokine release induced by mineral particles

The mechanisms for particle-induced health effects are not well understood, but inflammation seems to be of importance. Previously, we have shown that stone quarry particles with various mineral and metal content differed widely in potency to induce inflammatory cytokines (IL-6, IL-8 and TNF-alpha) in different types of lung cells. In this study we investigated if the observed cytokine responses were associated with the soluble or insoluble components of the stone particles and if there was a relationship between the differential cytokine release and generation of reactive oxygen species (ROS). Exposure of the human alveolar cell line A549 to the different particle leachates (pH 7.4 and 4.0) did not induce corresponding differential increases in the IL-8 release as observed with whole particles. Increase in ROS production, measured as dichlorofluorescein-fluorescence, was only demonstrated after exposure of A549 cells to the pH 4.0 extract from basalt. Furthermore, generation of ROS was found in neutrophils but not in A549 cells and primary macrophages after exposure to suspensions of the solid particles. However, no obvious differences in potency among the different particles were demonstrated. In summary, other mechanisms than particle-induced ROS formation seem to be responsible for the differential induction of IL-8. Furthermore, our findings indicate that the differential ability to induce IL-8 release in lung cells is attributed to the solid components of the stone particles.     

Methotrexate suppresses the interleukin-6 induced generation of reactive oxygen species in the synoviocytes of rheumatoid arthritis

Various cytokines and reactive oxygen species (ROS) play a fundamental role in the inflammatory and immunologic processes of rheumatoid arthritis (RA). Methotrexate (MTX) is one of the disease-modifying anti-rheumatic drugs and its effect may be partly due to the modulation of immunologic or inflammatory reactions by some cytokines. In the present study, we investigated the effects of MTX on the gene expression and synthesis of interleukin-6 (IL-6), and the proliferative activity and the production of ROS in the fibroblast-like synoviocytes (FLSs) obtained from the patient of RA. The expression or production of IL-6 was induced spontaneously, and augmented by the addition of recombinant human IL-6 or recombinant human IL-1 beta and TNF-alpha in FLSs. These spontaneous and augmented IL-6 expressions or productions were suppressed by treatment with low-concentration of MTX (1 microg/ml). Also, IL-6 stimulated the proliferation of FLSs, and this IL-6 driven proliferation was inhibited with the treatment of MTX or N-acetylcysteine (NAC, 1 mM). Furthermore, ROS production in FLSs was increased significantly by IL-6, and its effect was also abrogated in the presence of MTX or NAC. These results suggest that inflammatory reaction in the synovium of RA patients could be augmented by the autocrine or other cytokine-induced production of IL-6 with subsequent generation of ROS in the synoviocytes, and the modulations of IL-6 synthesis and ROS production may contribute to the therapeutic effects of MTX for RA.