广藿香叶挥发油抗炎作用机制实验研究

目的对广藿香叶挥发油抗炎机制进行实验研究。方法通过研究广藿香叶挥发油对角叉菜胶致大鼠足肿炎症模型组织和血清中前列腺素E2(PGE2)、丙二醛(MDA)、一氧化氮(NO)含量的影响,探讨广藿香叶抗炎作用机理。结果广藿香叶挥发油能明显降低大鼠角叉菜胶性炎症模型组织和血清中的PGE2和MDA的含量,能降低角叉菜胶致大鼠足肿炎症模型血清中NO的含量。结论广藿香叶挥发油抗炎作用机制与抑制炎症组织PGE2合成、减少炎症组织中MDA堆积,降低NO含量有关。     

退热解毒灵颗粒的解热抗炎作用

目的:研究退热解毒灵颗粒解热抗炎作用及抗炎机制。方法:40只SD大鼠随机分为空白对照组、模型对照组、退热解毒灵小剂量组(小剂量组)、退热解毒灵大剂量组(大剂量组)和阳性对照组,每组8只,观察在灌胃退热解毒灵颗粒6,15 g·kg-1后不同时间点的体温变化以及足肿胀度的变化,并观察退热解毒灵颗粒对角叉菜胶所致大鼠足肿胀炎症组织中前列腺素E2(prostaglandin E2,PGE2)、丙二醛(malondialdehyde,MDA)及一氧化氮(nitric oxide,NO)含量的影响。结果:大鼠背部皮下注射酵母5 h后体温明显升高(P<0.05),7 h大剂量组较模型组体温下降(P<0.05),8 h大、小剂量组较模型组体温下降(P<0.05),10 h大剂量组体温基本接近正常;大鼠右后足趾腱膜下注射角叉菜胶致炎1 h后大、小剂量组较模型组均对肿胀有明显的抑制(P<0.01,P<0.05),且在2 h作用效果最明显,抑制率分别为45.0%,35.0%;大剂量组能有效抑制大鼠足肿胀组织中PGE2、MDA及NO的含量;小剂量组能有效抑制大鼠足肿胀组织中NO的含量,大鼠足肿胀组织中PGE2、MDA的含量未见明显变化。结论:退热解毒灵颗粒有显著的解热抗炎作用,其抗炎机制可能与减少炎性组织中PGE2、NO的生成,抑制组织炎症中脂质过氧化产物MDA形成有关。     

桂产藿香蓟乙醇提取物抗炎作用及其机制研究

目的：探讨桂产藿香蓟乙醇提取物的抗炎作用及其作用机制。方法：采用二甲苯诱导小鼠耳廓肿胀,观察桂产藿香蓟乙醇提取物对二甲苯诱导小鼠耳廓肿胀的影响;采用角叉菜胶致小鼠足肿胀法,观察桂产藿香蓟乙醇提取物对角叉菜胶致小鼠足肿胀的影响,并分别测定小鼠炎性组织中丙二醛（MDA）、前列腺素E2（PGE2）和超氧化物歧化酶（SOD）的活性;采用角叉菜胶致大鼠足肿胀法,观察桂产藿香蓟乙醇提取物对角叉菜胶诱导大鼠足肿胀的影响,同时测定大鼠血清肿瘤坏死因子（TNF-α）、白介素-1β（IL-1β）和白介素-6（IL-6）的含量。结果：与空白对照组比较,桂产藿香蓟乙醇提取物能显著抑制小鼠耳廓肿胀及足肿胀（P〈0.05或0.01）,其中高、中、低剂量组（6.0,3.0,1.5g·kg-1）耳廓肿胀和足肿胀抑制率分别为29.24%,16.42%,11.21%和28.66%,18.79%,13.13%,并能降低小鼠炎足中炎性组织PGE2、MDA含量,提高其SOD活力（P〈0.05或0.01）;显著抑制大鼠的足肿胀（P〈0.05或0.01）,其中在3h时高、中、低剂量组（6.0,3.0,1.5g·kg-1）的足肿胀抑制率分别为43.69%,36.01%,23.29%,并能显著降低肿胀足大鼠血清TNF-α、IL-1β和IL-6的含量（P〈0.05或0.01）。结论：桂产藿香蓟乙醇提取物抗炎作用显著,其机制可能与清除氧自由基、减少炎症因子和致炎细胞因子的释放有关。     

羧胺三唑对角叉菜胶诱导急性炎症的抗炎作用

目的:探讨羧胺三唑(CAI)对角叉菜胶诱导的大鼠足爪急性炎症的预防作用及部分机制.方法:采用角叉菜胶诱导大鼠足爪急性炎症模型,测量足爪肿胀度以及足爪组织中髓过氧化物酶(MPO)、NO、前列腺素2(PGE2)、TNF-α、IL-1β、中性粒细胞趋化因子-1(CNCI-1)和诱导一氧化氮合酶(iNOS).结果:CAI能明显减轻大鼠足爪炎性肿胀,显著降低炎症足爪中MPO、NO、PGE2、TNF-α、IL-1β和CNCI-1含量及iNOS蛋白表达.结论:CAI可能通过减少炎症介质和中性粒细胞浸润来发挥对角叉菜胶诱导急性炎症的抗炎作用.     

虎杖提取物抗炎作用的实验研究

[目的]考察虎杖的乙酸乙酯提取物的抗炎作用及初步机制。[方法]分别以地塞米松、阿司匹林和环磷酰胺为阳性对照药,采用虎杖的乙酸乙酯提取物对小鼠和大鼠的多种炎症模型进行抑制实验。[结果]虎杖的乙酸乙酯提取物对角叉菜胶所致大鼠足跖肿胀有显著抑制作用(P<0.00 J);对小鼠、大鼠纸片法所致肉芽肿有显著抑制作用(P<0.01);对腹腔注射醋酸所致小鼠毛细血管通透性增高有显著抑制作用(P<0.01);对角叉菜胶所致小鼠肿胀足中炎症介质PGE2合成有显著抑制作用(P<0.001);对三硝基氯苯诱导的小鼠迟发性超敏反应有显著抑制作用(P<0.01);对肾上腺切除大鼠纸片法致肉芽肿(P<0.01)及角叉菜胶所致足跖肿胀(P<0.05)有显著抑制作用,但其抑制率均较正常大鼠相应实验抑制率有所降低。[结论]虎杖的乙酸乙酯提取物有一定的抗炎作用,其作用机制可能是抑制炎症介质PGE2的合成、抑制细胞免疫及与垂体-肾上腺皮质系统有关。     

广西甜茶提取物的抗炎作用研究

目的:研究广西甜茶提取物的抗炎作用。方法:观察广西甜茶提取物对巴豆油诱发小鼠急性耳肿胀和对血管通透性的抑制作用及角叉菜胶引起小鼠足跖肿胀的影响。利用Griess化学法检测小鼠腹腔巨噬细胞NO含量,RT-PCR检测一氧化氮合酶(iNOS)的表达。结果:广西甜茶提取物能显著抑制由巴豆油诱发的小鼠耳肿胀及血管通透性的增高,并对角叉菜胶引起的足跖肿胀也显示一定的抑制作用。广西甜茶提取物可抑制巨噬细胞NO生成和iNOSmRNA表达。结论:广西甜茶具有抗炎作用,其作用机制可能与抑制巨噬细胞NO生成和iNOSmRNA表达有关。     

马钱子总生物碱对实验性关节炎的影响

目的:观察马钱子总生物碱及士的宁对实验性关节炎的影响.方法:从马钱子中提取生物碱并进一步分离出番木鳖碱(士的宁)、总生物碱(除去部分士的宁)、非生物碱3个部分,观察各部分对角叉菜胶引起的大鼠足肿胀及棉球致肉芽组织增生的影响.结果:总生物碱部分能明显抑制大鼠足肿胀,第4,5,6小时抑制率分别为19.76%,30.02%,36.89%;明显抑制大鼠肉芽组织增生,抑制率37.60%,士的宁及非生物碱部分对上述炎症无明显影响.结论:总生物碱具有抗炎作用,强毒性成分士的宁为无效成分.     

侧柏总黄酮的抗炎作用

目的研究侧柏总黄酮的抗炎作用。方法以二甲苯致小鼠耳肿胀及角叉菜胶诱发大鼠足爪肿胀模型分别探讨侧柏总黄酮对小鼠耳片肿胀及大鼠足爪肿胀的抑制作用。测定大鼠足爪组织中一氧化氮 (NO)及前列腺素E2 (PGE2 )的含量。结果侧柏总黄酮在 12 5～ 5 0 0mg·kg-1内呈剂量依赖地抑制二甲苯诱导的小鼠耳肿胀 ,2 5 0mg·kg-1剂量能显著抑制大鼠足爪肿胀 ,其作用强于2 0mg·kg-1的地塞米松。侧柏总黄酮能抑制大鼠炎症足爪组织NO及PGE2 的生物合成。结论侧柏总黄酮具有较强的抗炎作用 ,其抗炎作用可能与抑制NO及PGE2 的生物合成或释放有关。     

小叶三点金提取物的抗炎作用及其机制研究

目的:研究小叶三点金提取物的抗炎作用及其机制,为该药用植物的进一步开发应用提供实验依据。方法:采用二甲苯、冰醋酸、角叉菜胶建立小鼠急性炎症模型,以地塞米松为阳性对照(0.005 g/kg),考察灌胃不同剂量小叶三点金提取物(50、30、15g/kg)对二甲苯致正常小鼠和肾上腺摘除小鼠耳肿胀、冰醋酸致正常小鼠腹腔毛细血管通透性增加、角叉菜胶致正常小鼠和肾上腺摘除小鼠足肿胀的抑制作用,并对角叉菜胶致炎模型肾上腺摘除小鼠足趾炎性部位中的丙二醛(MDA)、超氧化物歧化酶(SOD)、一氧化氮(NO)水平进行检测;另设空白组小鼠为对照(灌胃等体积水)。结果:与空白组比较,小叶三点金提取物高、中剂量组正常小鼠和肾上腺摘除小鼠耳肿胀度均显著降低,而耳肿胀抑制率显著升高;小叶三点金提取物各剂量组正常小鼠腹腔毛细血管通透性均显著降低;小叶三点金提取物高、中剂量组正常小鼠和肾上腺摘除小鼠足肿胀度均显著降低,而足肿胀抑制率显著升高(P<0.05或P<0.01);小叶三点金提取物高、中剂量组肾上腺摘除小鼠足趾炎性部位中MDA、NO水平均显著降低,SOD水平均显著升高(P<0.05)。结论:小叶三点金提取物对小鼠急性炎症有明显抑制作用;其抗炎作用机制与降低MDA、NO水平并提高SOD水平有关,且该抗炎作用的发挥不依赖于下丘脑-垂体-肾上腺轴系统。     

黄芩苷元对炎症反应的影响

目的研究以黄芩苷元为主要成分的黄芩提取物SBM抗炎作用,并初步探讨其作用机制。方法观察SBM体外对PGE2合成、COX-2酶活性,COX-2蛋白表达、p38蛋白磷酸化的影响;观察给药后对角叉菜胶诱导大鼠足肿胀、醋酸诱导的小鼠扭体反应的影响。结果SBM可抑制LPS诱导腹腔巨噬细胞PGE2合成、COX-2酶活性及COX-2蛋白表达;抑制ConA诱导的小鼠脾淋巴细胞p38蛋白磷酸化;并可抑制角叉菜胶诱导的大鼠足肿胀及醋酸诱导的小鼠扭体反应。结论黄芩提取物SBM可通过抑制淋巴细胞功能、炎症介质产生发挥抗炎作用。     

The ruthenium nitric oxide donor, [Ru(HEDTA)NO], inhibits acute nociception in mice by modulating oxidative stress,cytokine production and activating the cGMP/PKG/ATP-sensitive potassium channel signaling pathway

Nitric oxide plays an important role in various biological processes including antinociception. The control of its local concentration is crucial for obtaining the desired effect and can be achieved with exogenous nitric oxide-carriers such as ruthenium complexes. Therefore, we evaluated the analgesic effect and mechanism of action of the ruthenium nitric oxide donor [Ru(HEDTA)NO] focusing on the role of cytokines, oxidative stress and activation of the cyclic guanosine monophosphate/protein kinase G/ATP-sensitive potassium channel signaling pathway. It was observed that [Ru(HEDTA)NO] inhibited in a dose-dependent (1–10 mg/kg) manner the acetic acid-induced writhing response. At the dose of 1 mg/kg, [Ru(HEDTA)NO] inhibited the phenyl-p-benzoquinone-induced writhing response, and formalin- and complete Freund’s adjuvant-induced licking and flinching responses. Systemic and local treatments with [Ru(HEDTA)NO] also inhibited the carrageenin-induced mechanical hyperalgesia and increase of myeloperoxidase activity in paw skin samples. Mechanistically, [Ru(HEDTA)NO] inhibited carrageenin-induced production of the hyperalgesic cytokines tumor necrosis factor-α and interleukin-1β, and decrease of reduced glutathione levels. Furthermore, the inhibitory effect of [Ru(HEDTA)NO] in the carrageenin-induced hyperalgesia and myeloperoxidase activity was prevented by the treatment with ODQ (soluble guanylyl cyclase inhibitor), KT5823 (protein kinase G inhibitor) and glybenclamide (ATP-sensitive potassium channel inhibitor), indicating that [Ru(HEDTA)NO] inhibits inflammatory hyperalgesia by activating the cyclic guanosine monophosphate/protein kinase G/ATP-sensitive potassium channel signaling pathway, respectively. These results demonstrate that [Ru(HEDTA)NO] exerts its analgesic effect in inflammation by inhibiting pro-nociceptive cytokine production, oxidative imbalance and activation of the nitric oxide/cyclic guanosine monophosphate/protein kinase G/ATP-sensitive potassium channel signaling pathway in mice.     

Anti-inflammatory effect of propolis through inhibition of nitric oxide production on carrageenin-induced mouse paw edema

The anti-inflammatory effect of propolis was compared with that of diclofenac, a non-steroidal anti-inflammatory drug, and L-N(G)-nitro arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, using carrageenin-induced mouse paw edema. When administered 10 min prior to carrageenin injection, propolis (1 : 1000, 1 : 100, p.o.), diclofenac (12.5, 50 mg/kg, p.o.) and L-NAME (10, 100 mg/kg, s.c.) showed a significant anti-inflammatory effect. The anti-inflammatory effects of propolis and L-NAME were significantly inhibited by L-arginine, a precursor of nitric oxide, but not by D-arginine. In contrast, the anti-inflammatory effect produced by diclofenac was not inhibited by either D-arginine or L-arginine. These results indicate that the anti-inflammatory effect of propolis on mouse paw edema acts via the inhibition of nitric oxide production, similar to that of L-NAME but not diclofenac.     

Modulation of acute inflammation by endogenous nitric oxide.

The role of endogenous nitric oxide (NO) in acute inflammation was investigated using two inhibitors of NO synthase (NG-nitro-L-arginine methyl ester(L-NAME) and NG-monomethyl-L-arginine (L-NMMA)) as well as L- or D-arginine. The effect of test compounds was studied on the carrageenin-induced increase in vascular permeability in rat skin and in dextran- and carrageenin-induced paw oedema. Both L-NAME and L-NMMA dose dependently inhibited the increase in vascular permeability and oedema formation. L- but not D-arginine increased these inflammatory responses and reversed the inhibitory effects of L-NAME and L-NMMA. In dexamethasone-treated rats L-arginine enhanced the dextran-induced oedema and the early phase of carrageenin-induced oedema but did not modify the inhibition by dexamethasone of the late phase of carrageenin-induced oedema. These results suggest that endogenous NO is released at the site of acute inflammation and modulates oedema formation. Depending on the time course or on the type of inflammation, NO may be predominantly generated by the constitutive or by the inducible NO synthase.     

Discovery and biological evaluation of the novel naturally occurring diterpene pepluanone as antiinflammatory agent

From the whole plant of Euphorbia peplus L., a new diterpene based on a rare pepluane skeleton, named pepluanone (1), was isolated together with a known pepluane diterpene (2). The stereostructure of pepluanone was determined on the basis of an extensive NMR study, MS data, and chemical reaction. The ability of these compounds to act as antiinflammatory agents has been evaluated for the first time by in vivo tests on carrageenin-induced rat paw edema, an experimental model of acute inflammation. Comparison of the bioactivity of pepluanone and compound 2 in terms of chemical structure, evidenced the high efficiency of pepluanone and the absence of appreciable activity for compound 2, thus giving a first insight into the structure-activity relationship. Further in vitro experiments performed on pepluanone let us hypothesize that its activity could be explained in reducing the production of nitric oxide, prostaglandin E(2), and TNF-alpha by inhibiting the expression of inducible nitric oxide synthase, cyclooxygenase-2, and TNF-alpha mRNA through the down-regulation of NF-kappaB binding activity.     

Nuclear factor-κB activation mediates inducible nitric oxide synthase expression in carrageenin-induced rat pleurisy

We studied the involvement of nuclear factor-kappaB (NF-kappaB) in the regulation of inducible nitric oxide synthase expression in carrageenin-induced rat pleurisy. Injection of 0.2 ml of 1% lambda-carrageenin into the pleural cavity of male Wistar rats caused after 6 h: (a) exudate formation and leukocyte migration into the pleural cavity; (b) inducible NO synthase protein expression and accumulation of NO2- plus NO3- in pleural exudate; (c) increase in p50/p65 nuclear level as well as NF-kappaB/DNA binding activity. Treatment of rats with pyrrolidine dithiocarbamate (10, 30, and 100 mg/kg) and N-alpha-p-tosyl-L-lysine chloromethylketone (30 mg/kg), two inhibitors of NF-kappaB activation, given subcutaneously concomitantly with carrageenin, caused a significant inhibition of all the parameters assayed. These results suggest that in carrageenin-induced rat pleurisy the activation of NF-kappaB plays a key role in inducible NO synthase protein expression and in the development of inflammatory response.     

Influences of the endothelium and hypoxia on neurogenic transmission in the isolated pulmonary artery of the rabbit.

1. The effects of nitric oxide (10(-6) M), N omega-nitro-L-arginine methylester (L-NAME, 10(-4) M, an inhibitor of nitric oxide synthase), endothelium removal, hypoxia and selective alpha-adrenoceptor antagonists on responses to nerve electrical field-stimulation (EFS) were studied in the rabbit isolated pulmonary artery. 2. EFS induced frequency-dependent contractions which were abolished by prazosin (alpha 1-adrenoceptor antagonist) and unaffected by rauwolscine (alpha 2-adrenoceptor antagonist). EFS-induced responses were potentiated by L-NAME and inhibited by nitric oxide. The effect of L-NAME was reversed by the presence of L-arginine (2 x 10(-4) M), which had no effect on its own. In the presence of L-NAME, the EFS-induced responses were reduced by rauwolscine and the residual responses were abolished by prazosin. 3. Removal of the vascular endothelium increased the maximum contractile response to EFS but did not inhibit the ability of L-NAME to potentiate contractile responses to EFS. 4. Hypoxia inhibited the contractile response to EFS. This effect of hypoxia was also seen in the presence of L-NAME and in endothelium rubbed preparations. 5. In conclusion, the endothelium modulates EFS-induced contractions in the rabbit pulmonary artery. The contraction induced by EFS was inhibited by nitric oxide, but potentiated by the nitric oxide-synthase inhibitor, L-NAME. The effect of L-NAME was not mediated solely through the endothelium and revealed involvement of alpha 2-adrenoceptors in EFS-induced contraction. Hypoxia inhibited neurogenic responses in rabbit isolated pulmonary arteries.     

Irsogladine prevents monochloramine-induced gastric mucosal lesions by improving the decrease in mucosal blood flow due to the disturbance of nitric oxide synthesis in rats

The inhibitory effect of an anti-ulcer drug irsogladine [2,4-diamino-6-(2,5-dichlorophenyl)-s-triazine maleate] on monochloramine (NH(2)Cl)-induced gastric mucosal lesions and its mechanisms of action were clarified in rats. Irsogladine dose-dependently prevented the formation of gastric mucosal lesions induced by 60 mM NH(2)Cl. The mucosal protective effect of irsogladine was not influenced by capsaicin-sensitive sensory defunctionalization. On the other hand, its protective effect was diminished by the inhibitor of nitric oxide synthase N(G)-nitro-L-arginine methylester (L-NAME), but not by the inducible nitric oxide synthase selective inhibitor aminoguanidine. Irsogladine restored the NH(2)Cl-induced decrease in the gastric cGMP formation as an index of nitric oxide synthesis, while it alone had no influence on the cGMP formation in intact tissues. Pretreatment with L-NAME abolished the recovery of cGMP by irsogladine. Furthermore, irsogladine ameliorated the NH(2)Cl-induced decrease in gastric mucosal blood flow, which was also reversed by pretreatment with L-NAME. These findings suggest that the improvement of the decrease in mucosal blood flow subsequent to the disturbance of gastric nitric oxide synthesis is involved in the protective effect of irsogladine on gastric mucosal lesions caused by NH(2)Cl.     

Comparative effects of azelnidipine and other Ca2+-channel blockers on the induction of inducible nitric oxide synthase in vascular smooth muscle cells

Overproduction of nitric oxide by inducible nitric oxide synthase contributes to the progression of cardiovascular disease. We investigated the effects of azelnidipine and other Ca2+-channel blockers on nitric oxide production by cultured aortic smooth muscle cells isolated from Wistar rats and human umbilical vein endothelial cells (HUVECs), using the Griess reaction and oxyhemoglobin method. Release of lactic dehydrogenase (LDH) was measured to evaluate cell damage, and immunohistochemistry was performed to examine the expression of inducible nitric oxide synthase and nitrotyrosine protein. Azelnidipine and other Ca2+-channel blockers inhibited the release of nitric oxide induced by lipopolysaccharide plus interferon-gamma. Azelnidipine inhibited it most potently among the Ca2+-channel blockers tested (azelnidipine, amlodipine, nifedipine, diltiazem, verapamil, and nicardipine) at a concentration of 10 microM. Longer stimulation with these agents induced the expression of inducible nitric oxide synthase and nitrotyrosine, with an increase of lactic dehydrogenase release, whereas azelnidipine suppressed these changes. In human umbilical vein endothelial cells, azelnidipine enhanced basal nitric oxide production by endothelial nitric oxide synthase. In conclusion, azelnidipine potently inhibited the induction of inducible nitric oxide synthase and then nitric oxide production in vascular smooth muscle cells, while enhancing constitutive nitric oxide production by endothelial cells. Azelnidipine may inhibit nitrotyrosine expression and cell damage caused by overproduction of nitric oxide, suggesting a mechanism for its cardiovascular protective effect.     

In-vitro and in-vivo anti-inflammatory effect of oxyresveratrol from Morus alba L

The antioxidative effects of mulberroside A and oxyresveratrol obtained from Mori Cortex were examined. Mulberroside A and oxyresveratrol showed an inhibitory effect against FeSO4/H2O2-induced lipid peroxidation in rat microsomes and a scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radical. The anti-inflammatory effects of mulberroside A and oxyresveratrol using the carrageenin-induced model of inflammation were investigated in rats. Mulberroside A and oxyresveratrol significantly reduced paw edema. To investigate the mechanism of the anti-inflammatory action of these compounds, we examined the effects of oxyresveratrol on lipopolysaccharide (LPS)-induced responses in murine macrophage cell line RAW 264.7. Exposure of LPS-stimulated cells to oxyresveratrol inhibited nitrite accumulation in the culture medium. Oxyresveratrol also inhibited the LPS-stimulated increase of inducible nitric oxide synthase (iNOS) expression in a concentration-dependent manner; however, it had little effect on iNOS enzyme activity, suggesting that the inhibitory activity of oxyresveratrol is mainly due to the inhibition of iNOS expression rather than iNOS enzyme activity. Oxyresveratrol significantly inhibited LPS-evoked nuclear translocation of NF-kappaB and cyclooxygenase-2 (COX-2) activity in RAW 264.7 cells. The results suggest that the anti-inflammatory properties of oxyresveratrol might be correlated with inhibition of the iNOS expression through down-regulation of NF-kappaB binding activity and significant inhibition of COX-2 activity.