白藜芦醇抗炎作用机制的初步研究

 目的观察白藜芦醇的抗炎作用并对其抗炎机制进行初步探讨。方法采用切除双侧肾上腺大鼠足肿胀、冰醋酸致大鼠胸膜炎及角叉菜胶致大鼠气囊炎3种炎症模型,观察白藜芦醇对不同炎症模型的抗炎作用,并测量白细胞数量、蛋白质含量、前列腺素E₂(PGE₂)、一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)、超氧阴离子(O₂.-)含量变化。结果白藜芦醇高、中剂量组可减轻去双侧肾上腺大鼠足肿胀程度;减少了冰醋酸致大鼠胸膜炎渗出液量和蛋白质含量,使血清NO含量降低,SOD活性增强;抑制了角叉菜胶致大鼠气囊炎模型中白细胞游出及蛋白质渗出,降低PGE₂,NO,MDA,O₂.-生成量。结论白藜芦醇具有明显的抗炎作用,其抗炎作用不依赖于肾上腺的存在,可能与其降低血管通透性、抑制白细胞游出,抑制PGE₂、NO等炎症介质产生及增强清除氧自由基、抗脂质过氧化能力有关。     

蛇床子素对大鼠的抗炎作用和机制

目的:研究蛇床子素(Ost)的抗炎作用及机制.方法:用角叉菜胶诱导大鼠足肿模型.研究Ost对炎症的影响.NOS活性用NADPH黄递酶染色法,NO含量用Griess重氮化反应法,MDA用TBA荧光法测定,前列腺素类含量的测定通过紫外可见光分光光度法,经过0.5 mol/L氢氧化钾甲醇溶液,在50℃的温度中为异构后,在波长为278 nm处检测.结果:给角叉菜胶4小时后NO的含量明显提高,而蛇床子素各组对NO的含量有明显抑制作用.从单纯给角叉菜胶组NO为524.4±58.7 μmol/L到蛇床子素50 mg/kg、100 mg/kg NO的含量分别为461.5±68.8,353.8±111.0/μmol/L NO 2,表明对炎症因子NO的抑制作用呈剂量依赖性(P＜0.05).蛇床子素100 mg/kg对致炎大鼠NOS活性有明显抑制效应,其水平接近空白对照组.与此同时,结果显示蛇床子素也抑制致炎大鼠的脂质过氧化产物--MDA的产生.蛇床子素50,100 mg/kg显著抑制角叉菜胶致炎大鼠前列腺素类水平.结论:通过常见的实验方法表明蛇床子素具有抗炎作用,其抑制作用可能是通过抑制细胞钙内流和提高环鸟苷酸的水平而影响PG,NO,MDA等含量和cNOS的活性.     

柴郁汤的抗炎作用研究

目的:探讨柴郁汤的抗炎作用及其机理。方法:采用角叉菜胶诱发大鼠足肿胀模型和大鼠背部气囊模型、巴豆油混合致炎剂诱发小鼠耳肿胀模型以及鲁米诺化学发光法测定多形核白细胞(PMN)的发光强度。大鼠气囊炎性渗出液中前列腺素E2(prostaglandin E2,PGE2)、白细胞介素-6(interleukinI,L-6)和大鼠血清皮质醇用放射免疫法测定,一氧化氮(nitric oxide,NO)用硝酸还原酶法测定。结果:柴郁汤对大鼠足肿胀有显著抑制作用,能明显减少气囊炎性渗液中PGE2I、L-6、NO,对气囊模型大鼠血清皮质醇含量无明显影响,柴郁汤药物血清和姜黄素能明显抑制激活的PMN化学发光。结论:柴郁汤有明显的抗炎作用,其抗炎作用机制部分在于抑制PGE2、NOI、L-6的产生,抑制PMN产生氧自由基,但与影响下丘脑-垂体-肾上腺皮质轴无关。     

蜂胶的外用抗炎作用实验研究

目的:观察蜂胶的外用抗炎作用.方法:昆明种小鼠60只,随机分为模型对照组(3.0 g·kg-1吐温80)、阳性药布洛芬乳膏组(4.0 g·kg-1),蜂胶高、中、低剂量组(0.72,0.24,0.08 g·kg-1),每组12只采用二甲苯致小鼠耳廓肿胀模型,检测小鼠血清中前列腺素E2(PGE2),一氧化氮(N0)含量,观察蜂胶对二甲苯致小鼠耳廓肿胀炎症模型的影响;Wistar大鼠60只,分组同上,模型对照组(3.0 g·kg-1吐温80),布洛芬乳膏组(3.0g·kg-1),蜂胶高、中、低剂量组(0.20,0.10,0.05 g·kg-1),采用角叉菜胶致大鼠足跖肿胀模型,检测大鼠炎症组织中PGE2含量,观察蜂胶对角叉菜胶致大鼠足跖肿胀炎症模型的影响.结果:蜂胶能明显抑制二甲苯导致的小鼠耳肿胀(P＜0.05);能明显减轻角叉菜胶导致的大鼠足肿胀(P＜0.05);能显著降低小鼠血清中PGE2,NO含量(P＜0.01);能显著降低大鼠炎症组织中PGE2含量(P＜0.01).结论:蜂胶外用具有明显的抗炎作用.     

紫花地丁抗炎作用及机制研究

目的:观察紫花地丁水提物及醇提物的抗炎作用,并分析其抗炎机制.方法:取ICR小鼠随机分模型对照组,布洛芬组(0.08 g·kg-1),紫花地丁水提物高、低剂量(9.0,3.0 g·kg-1)组,紫花地丁醇提物高、低剂量(9.0,3.0 g·kg-1)组,每天1次,连续ig给药7d,末次给药30 min后,采用二甲苯致小鼠耳肿胀法和角叉菜胶致小鼠足肿胀法观察抗炎作用,并采用紫外分光光度法和酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测炎性组织中前列腺素E2(PGE2,prostaglandin E2)和血清中肿瘤坏死因子(tumor necrosis factor-alpha,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)的含量,探讨抗炎作用机制.结果:紫花地丁水提物及醇提物各剂量组(9.0,3.0 g·kg-1)对二甲苯致小鼠耳肿胀及角叉菜胶致小鼠足肿胀均具有显著的抑制作用,与模型对照组比较,差异均具有统计学意义(P＜0.05～P＜0.01);并可不同程度降低角叉菜胶致炎小鼠血清TNF-α,IL-1β及炎性组织中PGE2含量,高剂量组(9.0 g·kg-1)的作用显著,与模型对照组的差异具有统计学意义(P＜0.01).结论:紫花地丁水提物及醇提物均具有显著的抗炎作用,其抗炎作用机制可能与降低TNF-α,IL-1β及PGE2的表达有关.     

四乙酰葛根素抗炎作用及机制研究

目的:探讨四乙酰葛根素的抗炎作用并初步研究其作用机制.方法:将雄性SD大鼠随机分为正常对照组、模型组和四乙酰葛根素高、中、低剂量( 80,40,20 mg· kg-1)组以及吲哚美辛(5 mg·kg-1)组,四乙酰葛根素尾iv给药4d,末次给药30 min后予以造模,观察四乙酰葛根素对角叉菜胶所致大鼠急性足肿胀、炎性组织中前列腺素E2( PGE2)和血清中肿瘤坏死因子α( TNF-α)、白介素1β(IL-1β)含量的影响.结果:四乙酰葛根素(80 mg·kg-1)能明显抑制模型大鼠的足肿胀(P＜0.01),降低肿胀足爪组织中PGE2(P＜0.05)及血清中TNF-α(P＜0.05),IL-1β(P ＜0.05)的含量.结论:四乙酰葛根素可能通过影响炎性因子释放而起到抑制大鼠炎性足肿胀的作用.     

培养雪莲总黄酮对巨噬细胞炎性介质一氧化氮和前列腺素E_2的影响

目的:研究培养雪莲总黄酮对炎性介质一氧化氮(NO),前列腺素E2(PGE2)分泌以及一氧化氮合酶(iNOS),环氧酶-2(COX-2)mRNA表达的影响,探讨其抗炎机制。方法:用Griess法测定培养雪莲总黄酮对Raw264.7细胞NO的影响,放射免疫法测定培养雪莲总黄酮对Raw264.7细胞PGE2含量;逆转录聚合酶链式反应方法检测Raw264.7细胞iNOS,COX-2mRNA表达情况。结果:培养雪莲总黄酮可明显降低Raw264.7细胞上清中NO,PGE2含量,同时,降低iNOS,COX-2 mRNA表达。结论:培养雪莲总黄酮的抗炎作用与抑制炎性介质NO,PGE2分泌有关,抑制iNOS,COX-2 mRNA表达从而降低炎性介质NO,PGE2分泌可能是培养雪莲总黄酮的抗炎作用的机制之一。     

祖师麻醋酸乙酯提取物体外抗炎作用及其机制

目的研究祖师麻醋酸乙酯提取物(EAEDGC)的体外抗炎作用及其机制。方法用γ-干扰素(IFN-γ)和脂多糖(LPS)协同制备小鼠RAW264.7细胞炎症模型,Griess反应测定细胞上清液中NO生成量,MTT法测定细胞活力,三价铁还原抗氧化能力测试(FRAP assay)测定细胞总抗氧化能力,RT-PCR检测诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)、血红素加氧酶-1(HO-1)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)mRNA的表达;Western blotting检测iNOS、COX-2、HO-1、p-ERK蛋白表达水平。结果 EAEDGC以剂量依赖方式抑制细胞上清液中NO的生成,提高细胞总抗氧化能力,下调iNOS、IL-1β、IL-6、TNF-αmRNA及iNOS、p-ERK蛋白的表达,上调HO-1 mRNA和蛋白表达,同时不影响细胞生长;但对COX-2 mRNA和蛋白表达影响不大。结论 EAEDGC部分通过丝裂原活化蛋白激酶(MAPK)中的细胞外信号调节激酶(ERK)信号转导通路抑制iNOS基因和蛋白的表达,从而抑制NO的生成,提高细胞总抗氧化能力,同时下调IL-1β、IL-6、TNF-α炎症介质和上调抗炎介质HO-1的表达,进而发挥抗炎作用。     

赤雹根总皂苷抗炎作用研究

目的:观察赤雹根总皂苷的抗炎活性,为其研究开发提供实验依据.方法:采用二甲苯致小鼠耳肿胀法,角叉菜胶致小鼠足肿胀法,小鼠棉球肉芽肿法,评价赤雹根总皂苷的抗炎作用.二甲苯耳肿胀法和角叉菜胶致足肿胀法,各组小鼠每日灌胃给药1次,连续3d,末次给药后30 min,复制模型.小鼠棉球肉芽肿实验,棉球植入术后小鼠连续灌胃给药7d.实验动物均分为赤雹根总皂苷高、中、低( 240,120,60 mg·kg-1)剂量组、阿司匹林(100 mg·kg-1)组、模型组(生理盐水).结果:赤雹根总皂苷高、中、低剂量组小鼠的耳肿胀度[(1.46±0.19),(1.68±0.15),(1.99±0.17) mmg]较模型组(2.31±0.24)mg明显降低(P＜0.01);赤雹根总皂苷各剂量组小鼠足肿胀度[(28.7±5.03),(33.8±4.65),(43.33±4.98) mg]较模型组(55.18±2.96) mg明显降低(P＜0.01);赤雹根总皂苷各组小鼠肉芽肿质量(5.08±0.37,6.76±0.44,8.66±0.55) mg较模型组(12.05 ±1.22) mg明显降低(P＜0.01),且各实验中赤雹根总皂苷高和(或)中剂量效果优于阿司匹林(P＜0.05～0.01).角叉菜胶致小鼠足肿胀实验中,赤雹根总皂苷各组小鼠血清前列腺素E2(PGE2)吸光度值(0.193±0.022,0.219±0.035,0.250±0.037)和一氧化氮(N0)含量(5.85±3.50,9.62±3.61,16.79±3.14)μmol·L-1较模型组PGE2吸光度值(0.545±0.06),NO含量(23.02±3.18)μmol· L-1明显降低(P＜0.01).结论:赤雹根总皂苷有显著的抗炎作用,其抗炎作用与抑制炎症介质PGE,和NO的合成释放有关.     

青蒿素和二氢青蒿素的抗炎作用及机制

目的:研究并比较青蒿素和二氢青蒿素的抗炎作用及其分子机制.方法:用LPS刺激小鼠单核巨噬细胞RAW 264.7释放TNF-α,IL-6,NO等炎症介质,评价药物对巨噬细胞释放以上炎症介质的抑制作用.以ELISA法检测TNF-α,IL-6的含量,以Griess法检测NO的含量,同时以MTF法评价细胞毒性.Western blot法检测iNOS,COX-2及内参蛋白β-actin水平.比色法检测COX-2酶活性.结果:二氢青蒿素在12.5～100μnol·L-1可明显抑制LPS诱导小鼠巨噬细胞RAW 264.7释放TNF-α,IL-6及NO,并呈现良好的剂量依赖关系.青蒿素仅对IL-6具有一定程度的抑制作用.结论:二氢青蒿素通过下调iNOS蛋白表达,抑制巨噬细胞释放炎症因子TNF-α,IL-6和炎症介质NO发挥抗炎活性,青蒿素可能通过代谢为二氢青蒿素发挥抗炎作用.     

Anti-inflammatory and antitumoural effects of Uncaria guianensis bark

Ethnopharmacological relevance

Uncaria guianensis (Aublet) Gmell (Rubiaceae) is a medicinal plant from the jungles of South and Central America, used to treat cancer, arthritis, diabetes, and inflammation. Evaluate the anti-inflammatory and anti-tumor effects of Uncaria guianensis preparations.

Materials and methods

Bio-guided fractionation of a hydroethanolic extract of Uncaria guianensis was performed, evaluating the fractions and subfractions for their effect on inflammatory mediators, tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and prostaglandin E₂ (PGE₂) by ELISA and nitric oxide (NO) by the Griess reaction in cultured supernatant from RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). The expression of cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS) and inhibitor of κB (IκB) were investigated in RAW 264.7 macrophages by flow cytometry. The activity of NF-κB in HeLa cells transfected with a luciferase reporter system was determined. The effect of Uncaria guianensis on the inflammatory response in vivo was assessed in BALB/c mice stimulated with LPS, on rat paw oedema induced by carrageenan, and on tumour growth and lung metastasis in BALB/c mice inoculated with 4T1 mammary tumour cells. Immune cell infiltrates and inflammatory mediators were evaluated in the tumour by immunohistochemistry.

Results

Sub-fraction Ug AIV inhibited, to varying degrees, NO, TNF-α, IL-6 and PGE₂ production by macrophages in vitro (30 μg/ml) and in the serum of LPS-challenged mice (5 mg/kg). Macrophage expression of Cox-2 was inhibited (35%), IκB degradation was completely inhibited and NF-κB activation was inhibited (70%) by Ug AIV at 30 μg/ml. Ug AIV decreased paw oedema by 86% (5 mg/kg) and serum NO and TNF-α by 45% and 65% respectively. Ug AIV reduced 4T1 mammary tumour growth by 91% on day 33 post-inoculation as well as the levels of serum NO, IL-6 and TNF-α in the same animals. Ug AIV decreased the number of tumour-infiltrating T lymphocytes, macrophages and neutrophils as well as the number of cells positive for COX-2, iNOS, IL-6, TNF-α and p65.

Conclusions

As Ug AIV was not cytotoxic for tumour cells or macrophages, its anti-tumour effect may be due to a reduction in pro-tumoural inflammatory processes in the tumour microenvironment, possibly mediated through NF-κB.     

Anti-inflammatory mechanism of Kaempferia parviflora in murine macrophage cells (RAW 264.7) and in experimental animals

Ethnopharmacological relevance

The rhizomes of Kaempferia parviflora Wall. ex Baker have been used in Thailand for treatment of gout, apthous ulcer, peptic ulcer and abscesses.

Aim of the study

In our previous study, the crude ethanol extract of Kaempferia parviflora and its compound (5, 5-hydroxy-3,7,3′,4′-tetramethoxyflavone), was reported to show nitric oxide (NO) inhibition in RAW 264.7 cells. The present study is thus investigated the anti-inflammatory mechanism of Kaempferia parviflora extract and compound 5 against inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA expressions.

Materials and methods

The extract of Kaempferia parviflora and its compound were tested against NO and prostaglandin E₂ (PGE₂) releases using RAW264.7 cells as well as studied on anti-inflammatory activity in carrageenan-induced rat paw edema and acute toxicity in mice.

Results

The results revealed that the ethanol extract of Kaempferia parviflora markedly inhibited PGE₂ release with an IC₅₀ value of 9.2 μg/ml. This plant extract and compound 5 also suppressed mRNA expression of iNOS in dose-dependent manners, whereas COX-2 mRNA expression was partly affected. According to the in vivo study, chloroform and hexane fractions greater decreased rat paw edema than ethanol, ethyl acetate and water fractions.

Conclusion

The mechanisms for anti-inflammatory activity of Kaempferia parviflora and compound 5 are mainly due to the inhibition of iNOS mRNA expression but partly through that of COX-2 mRNA.     

姜黄消痤搽剂对大、小鼠炎症组织的抗炎作用

目的:研究姜黄消痤搽剂的抗炎机制。方法:注射卵白蛋白(ovalbumim,OA)致小鼠足肿胀,观测姜黄消痤搽剂1.35,0.90,0.45 mg.kg-1外用,3次/d,连续4 d,对OA引起足肿胀的影响,检测炎症组织中前列腺素E2(PGE2)的含量;福氏完全佐剂(FCA)致大鼠关节炎,1.35,0.90,0.45 mg.kg-1外用,2次/d,连续30 d,检测致炎部位皮肤和皮下组织的病理改变和白细胞介素(IL-2)含量。结果:姜黄消痤搽剂可显著抑制OA引起足肿胀,并能降低炎症组织中PGE2的含量;对OA致敏激发皮炎皮损组织有明显治疗作用;可减轻FCA所致炎症反应并降低炎症组织中IL-2的含量。结论:姜黄消痤搽剂可能通过抑制PGE2,IL-2的产生而发挥抗炎作用。     

醋制老瓜头镇痛抗炎活性研究

目的: 研究醋制老瓜头二氯甲烷、乙酸乙酯、正丁醇层萃取物抗炎、镇痛活性,明确活性部位。 方法: 将小鼠分为11组,分别为:阳性对照组、模型组、醋制老瓜头二氯甲烷萃取物,乙酸乙酯萃取物,正丁醇萃取物分别设低、中、高剂量组(按生药量计为400,800,1 600 mg·kg^-1),给药3 d后进行实验,以热板法、醋酸扭体法为镇痛实验模型,二甲苯致小鼠耳肿胀、角叉菜胶致小鼠足肿胀、小鼠炎性组织前列腺素E₂(PGE₂)含量为抗炎实验模型,进行活性测定。 结果: 镇痛实验,热板法模型中醋制老瓜头各萃取层与模型组比较能显著延长热板所致的小鼠舔后肢的时间,扭体法模型中,醋制老瓜头各萃取层能显著减少醋酸致小鼠扭体反应的次数,并呈现一定的药物依赖性(P<0.05,P<0.01);抗炎实验,二甲苯致耳肿胀模型中醋制老瓜头各萃取层与模型组比较能不同程度地抑制二甲苯致小鼠耳肿胀,角叉菜胶致鼠足肿胀模型中醋制老瓜头各萃取层能不同程度抑制角叉菜胶致鼠足肿胀,各萃取层能显著降低小鼠炎症组织中PGE₂含量(P<0.05,P<0.01)。 结论: 二氯甲烷层为醋制老瓜头镇痛、抗炎活性最强部位,醋制降毒后仍有较强生物活性,表现为延长小鼠舔后肢的时间,抑制二甲苯致小鼠耳肿胀、角叉菜胶致足肿胀并显著降低小鼠炎症组织中PGE₂含量,其抗炎作用可能与PGE₂相关。     

鸡鸣散镇痛抗炎作用及其机理的时间药理学实验研究

目的:观察鸡鸣散(Ji-Ming-San,JMS)对啮齿类动物的毒性及镇痛抗炎作用,探讨其机制及昼夜节律性差异,验证前人关于JMS择时用药的合理性。方法:Bliss法计算JMS半数致死量(LD50)及相同剂量下JMS对小鼠急性死亡率的昼夜差异;热板法、扭体法观察JMS镇痛作用,硝酸还原酶法测定小鼠血清及脑组织一氧化氮(NO)含量及其昼夜差异;耳廓肿胀法和足跖肿胀法观察JMS抗炎作用,紫外分光光度计法测炎症组织前列腺素E2(PGE2)含量及其昼夜差异。结果:JMS对小鼠的急性毒性呈显著的用药时间依赖性,白昼毒性大于夜间;JMS对小鼠模型有良好的镇痛作用,呈昼夜节律性,并与其降低小鼠血清、脑组织NO含量的昼夜差异相吻合;JMS具有良好的抗炎作用,呈昼夜差异性,并与其降低炎性组织中PGE2含量的昼夜差异相吻合。结论:JMS具有良好的镇痛、抗炎作用,并呈昼夜节律性,镇痛作用可能与其降低NO含量有关,抗炎作用可能与其降低PGE2含量有关。     

Anti-inflammatory effect of bee venom on antigen-induced arthritis in rabbits: Influence of endogenous glucocorticoids

Aim of the study

This study assessed the involvement of endogenous glucocorticoids (GCs) in the anti-arthritic properties of bee venom (BV) on antigen-induced arthritis (AIA) in rabbits.

Materials and methods

BV (1.5–6 μg/kg/day) was injected for 7 days before AIA induction, whereas the control group received sterile saline. The total and differential leukocyte count, PGE₂ levels in synovial fluid and synovial membrane cell infiltrate were evaluated. The contribution of GCs to BV action was assessed in rabbits treated with BV plus metyrapone, an inhibitor of GC synthesis, or RU-38 486, a steroid antagonist.

Results

Treatment with BV (1.5 μg/kg/day) reduced the leukocyte count and PGE₂ level (18571 ± 1909 cells/mm³ and 0.49 ± 0.05 ng/mL, respectively) as well as the cellular infiltrate compared with the control group (40968 ± 5248 cells/mm³ and 2.92 ± 0.68 ng/mL, p < 0.05). The addition of metyrapone to BV treatment completely reversed the inhibition of AIA, whereas RU-38 486 was ineffective.

Conclusion

Our data show that bee venom treatment prevents the development of antigen-induced arthritis in rabbits through the action of GCs.