A novel property of propolis (bee glue): anti-pathogenic activity by inhibition of N-acyl-homoserine lactone mediated signaling in bacteria

Ethnopharmacological Relevance

An alternative approach to antibiotics is the development of anti-pathogenic agents to control the bacterial virulome. Such anti-pathogenic agents could target a phenomena known as quorum sensing (QS).

Materials and methods

Six bacterial N-acyl-homoserine lactone (AHL)-dependent bioreporter strains were used to evaluate if bee hive glue also known as propolis contains constituents capable of inhibiting QS-controlled AHL signaling. In addition, the effect of propolis on the QS-dependent swarming motility was evaluated with the opportunisitic pathogen, Pseudomonas aeruginosa.

Results

Differences in the propolis tincture samples were identified by physiochemical profiles and absorption spectra. Propolis tinctures at 0.0005% (v/v) that do not affect bacteria biosensor growth or the reporter system monitored were exposed to biosensors with and without the addition an AHL. No AHL signal mimics were found to be present in the propolis tinctures. However, when propolis and an inducer AHL signal were together exposed to five Escherichia coli and a Chromobacterium violaceum biosensor, propolis disrupted the QS bacterial signaling system in liquid- and agar-based bioassays and in C₁₈ reverse-phase thin-layer plate assays. Swarming motility in the opportunistic pathogen, Pseudomonas aeruginosa PAO1 and its AHL-dependent LasR- and RhlR-based QS behaviors were also inhibited by propolis.

Conclusions

Together, we present evidence that propolis contain compounds that suppress QS responses. In this regard, anti-pathogenic compounds from bee harvested propolis could be identified and isolated and thus will be valuable for the further development of therapeutics to disrupt QS signaling systems which regulate the virulome in many pathogenic bacteria.     

Effects of thymoquinone (volatile oil of black cumin) on rheumatoid arthritis in rat models

Many studies have been carried out in recent years on the pharmacological effects of nigella sativa seeds that have uncovered their antiinflammatory and immunological effects. The objective of this study was to explore the antiinflammatory effects of thymoquinone on arthritis in rat models. Rats with arthritis induced by Freund's incomplete adjuvant were assigned into five groups: group 1: controls 0.9% NaCl (n = 7); group 2: 2.5 mg/kg thymoquinone (n = 7); group 3: 5 mg/kg thymoquinone (n = 7); group 4: Bacilli Chalmette Guerin (BCG) 6 x 10(5) CFU (n = 7); group 5: methotrexate 0.3 mg/kg (n = 7). Signs of inflammation on the claw and radiological signs were searched for and TNF-alpha and IL-1beta were measured. The results of control and other groups were compared. As a result, thymoquinone, confirmed clinically and radiologically, suppressed adjuvant-induced arthritis in rats.     

Effects of thymoquinone (volatile oil of black cumin) on rheumatoid arthritis in rat models

Many studies have been carried out in recent years on the pharmacological effects of Nigella sativa seeds that have uncovered their antiinflammatory and immunological effects. The objective of this study was to explore the antiinflammatory effects of thymoquinone on arthritis in rat models. Rats with arthritis induced by Freund's incomplete adjuvant were assigned to five groups: group 1: controls 0.9% NaCl (n = 7); group 2: 2.5 mg/kg thymoquinone (n = 7); group 3: 5 mg/kg thymoquinone (n = 7); group 4: Bacilli Chalmette Guerin (BCG) 6 x 105 CFU (n = 7); group 5: methotrexate 0.3 mg/kg (n = 7). Signs of inflammation on the claw and radiological signs were searched for and TNF-alpha and IL-1beta were measured. The results of the control and other groups were compared. As a result, thymoquinone, confirmed clinically and radiologically, suppressed adjuvant-induced arthritis in rats.     

Chemical composition of the ethanolic propolis extracts and its effect on HeLa cells

Ethnopharmacological relevance

Propolis is a resinous hive product collected by honeybees from various plant sources. It is widely used in traditional medicine and is reported to have a broad spectrum of pharmacological effects (antibacterial, antihepatoxic, antioxidative, anti-inflammatory, etc.). Thus the aim of this study was to assess cytotoxic effect of various ethanol propolis extractions on the cervical tumor cell line (HeLa) and compare it with their phenolic acids and flavonoids composition.

Materials and methods

Twenty samples of raw propolis were collected from 17 localities of Croatia (I-XVII), 2 of Bosnia and Hercegovina (XVIII, XIX) and 1 of Macedonia (XX). Reverse phase HPLC was used to determine the chemical composition of polyphenols. Biological experiments were carried out in vitro on cervix adenocarcinoma cell line (HeLa).

Results

Phenolic acids (ferulic acid, p-coumaric acid, caffeic acid) and flavonoids (tectochrysin, galangin, pinocembrin, pinocembrin-7-methylether, chrysin, apigenin, kaempferol, quercetin) have been determined using HPLC analysis in 20 ethanolic propolis extracts. All samples contain tectochrysin in ranges of 0.1988 mg/g (XVIII) to 1.2004 mg/g (III), while caffeic acid and quercetin have not been found. Flavonoid that is most abundant is galangin in ranges from 0.3706 mg/g (XVII) to 47.4879 mg/g (IX). The samples of propolis numbers I, VI and X applied in the investigated concentration range manifested significant reduction of cell growth. GI₅₀ value as indicator of cytotoxicity among propolis samples showed that propolis number VII is the most effective (GI₅₀ = 76 μg/ml) followed by propolis nos. XV, XVIII and I.

Conclusion

Antiproliferative and cytotoxic effect of propolis on the HeLa cells is not correlating with the concentration of particular components but on establishing the possible synergistic antiproliferative activity of individual phenolic acid and flavonoids. Differences in the chemical composition lead to diversity in biological activity of propolis samples.     

蜂胶对大鼠佐剂性关节炎的作用及其机制的研究

 目的观察蜂胶液对佐剂性关节炎的作用及其机制。方法采用大鼠佐剂性关节炎模型进行观察比较蜂胶水提液和蜂胶醇提液的抗炎免疫作用。结果蜂胶醇提液和水提液均能够减轻局部组织的炎症反应,抑制足跖肿胀。通过测定大鼠关节炎渗出液中IFN-γ,IL-2,PGF₂和IL-6含量,发现蜂胶液对IL-6和PGE₂的产生有显著的抑制作用,而对IL-2和IFN-γ无明显影响。结论蜂胶具有良好的抗炎作用,其作用机制可能是通过抑制单核巨嗜细胞的活化和分化发挥抗炎作用。     

清络通痹颗粒对佐剂性关节炎大鼠滑膜细胞的影响

目的探讨清络通痹颗粒对佐剂性关节炎大鼠滑膜细胞的影响.方法建立佐剂性关节炎(AIA)大鼠模型,同时设正常对照组.自第7天开始给药,持续21 d.取滑膜,检测滑膜细胞增殖能力和滑膜细胞分泌IL-1的量.结果清络通痹颗粒7.2、14.4 g/kg可明显抑制AIA大鼠的滑膜细胞增殖反应(P＜0.01).结论一定剂量的清络通痹颗粒可抑制AIA大鼠滑膜细胞的过度增生,这与其调节与滑膜增生相关的细胞因子有关.     

Picrorhiza kurroa Inhibits Experimental Arthritis Through Inhibition of Pro‐inflammatory Cytokines,Angiogenesis and MMPs

The present study investigates the anti‐arthritic activity of Picrorhiza kurroa (PK), on formaldehyde and adjuvant‐induced arthritis (AIA) in rat. Administration of Picrorhiza kurroa rhizome extract (PKRE) significantly inhibited joint inflammation in both animal models. In AIA‐induced arthritic rat, treatment with PKRE considerably decreased synovial expression of interleukin‐1β (IL‐1β), interleukin‐6 (IL‐6), tumor necrosis factor receptor‐1 (TNF‐R1) and vascular endothelial growth factor as compared with control. The anti‐arthritic activity was found to be well substantiated with significant suppression of oxidative and inflammatory markers as there was decreased malonaldehyde, Nitric oxide, tumor necrosis factor alpha levels accompanied with increased glutathione and superoxide dismutase, catalase activities. Additionally, PKRE significantly inhibited the expression of degrading enzymes, matrix metalloproteinases‐3 and matrix metalloproteinases‐9 in AIA‐induced arthritic rat. Histopathology of paw tissue displayed decreased inflammatory cell infiltration as compared with control. Taken together, these results demonstrated the anti‐arthritic activity of PKRE against experimental arthritis, and the underlying mechanism behind this efficacy might be mediated by inhibition of inflammatory mediators and angiogenesis, improvement of the synovium redox status and decreased expression of matrix metalloproteinases. Copyright © 2015 John Wiley & Sons, Ltd.     

川芎嗪的抗炎作用与组胺、5-羟色胺的关系

目的观察川芎嗪的抗炎作用与组胺、5-羟色胺的关系.方法采用荧光法测定炎性渗出物中组胺、5-羟色胺的含量,用豚鼠离体回肠收缩实验观察川芎嗪对组胺、5-羟色胺的作用.结果川芎嗪能显著降低大鼠炎性渗出物中组胺、5-羟色胺的含量,显著拮抗组胺、5-羟色胺所致豚鼠离体回肠的收缩.结论川芎嗪的抗炎作用可能与其抑制组胺、5-羟色胺的合成或释放,及对组胺、5-羟色胺的直接拮抗作用有关.     

瑶药风湿骨痛喷雾剂对佐剂性关节炎大鼠的治疗作用

目的：通过瑶药风湿骨痛喷雾剂（ROS）对佐剂性关节炎（AA）大鼠的影响,得出其治疗效果,从而为ROS在临床上的应用提供理论依据。方法：参照文献完全弗氏佐剂造模方法致大鼠足趾出现继发性肿大,观察ROS高剂量（12.60 g·kg-1）、中剂量（6.30 g·kg-1）、低剂量（3.15 g·kg-1）三种剂量对AA大鼠继发性足趾肿大的影响,以及对细胞因子IL-1,IL-6,TNF-α,RF和炎症介质PGE2含量的影响。结果：ROS高剂量（12.60 g·kg-1）、中剂量（6.30 g·kg-1）、低剂量（3.15 g·kg-1）对AA大鼠继发性足趾肿大都有显著性作用,同时对IL-1、IL-6、TNF-α、RF和PGE2有抑制的作用或趋势。结论：ROS能够抑制AA大鼠足肿胀度,降低关节炎指数（AI）。     

基于NF-κB通路探讨杜仲皮、叶醇提物对胶原诱导型关节炎大鼠炎症性骨破坏的影响

目的 研究杜仲皮、叶醇提物对胶原诱导型关节炎(collagen-induced arthritis,CIA)大鼠骨破坏的治疗作用及机制.方法 随机选取6只雌性Wistar大鼠作为对照组,其余大鼠于背部多点及尾根部sc0.1 mL牛Ⅱ型胶原乳剂建立CIA模型,第14天将CIA大鼠随机分为模型组,杜仲皮醇提物低、高剂量(2...     

Effects of combining extracts (from propolis or Zingiber officinale) with clarithromycin on Helicobacter pylori

Propolis and Zingiber officinale have been shown to be specifically targeted against Helicobacter pylori strains, to possess antiinflammatory, antioxidant and antitumoral activity and to be used in traditional medicine for the treatment of gastrointestinal ailments. Considering that these natural products could potentially serve as novel therapeutic tools also in combination with an antibiotic, the aim of this work was to evaluate their effect when combined with clarithromycin on clinical H. pylori isolates (n = 25), characterized in respect to both clarithromycin susceptibility and the presence of the cagA gene. The results showed that the combinations of propolis extract + clarithromycin and Z. officinale extract + clarithromycin exhibited improved inhibition of H. pylori with synergistic or additive activity. Interestingly, the susceptibility to combinations was significantly independent of the microbial clarithromycin susceptibility status. Only one H. pylori strain showed antagonism towards the Z. officinale extract + clarithromycin combination. The data demonstrate that combinations of propolis extract + clarithromycin and Z. officinale extract + clarithromycin have the potential to help control H. pylori-associated gastroduodenal disease.     

电针、手针“足三里”对慢性关节炎大鼠血清炎性细胞因子含量的影响

目的:观察不同针刺方法对慢性佐剂性关节炎大鼠血清中致炎类因子白介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和抗炎类因子白介素2(IL-2)、白介素4(IL-4)及白介素10(IL-10)含量的影响,比较不同针刺方法的效应。方法:将SD大鼠随机分为正常组、模型组、电针单穴组、电针双穴组和手针单穴组,每组12只。大鼠右后肢足跖皮内注射0.1mL弗氏完全佐剂制造佐剂性关节炎模型。针刺各组于造模第11天起取"足三里"穴进行治疗,每次30min,每周治疗2次,共4周。以酶联免疫法检测大鼠血清中IL-1β、TNF-α、IL-2、IL-4及IL-10含量。结果:与正常组相比,模型组大鼠血清中IL-1β、TNF-α、IL-4及IL-10的含量均显著升高(P<0.001)。与模型组相比,各治疗组大鼠血清中致炎类因子IL-1β和TNF-α含量明显降低(P<0.001,P<0.05),抗炎类因子IL-2含量明显升高(P<0.05);各治疗组大鼠血清中的IL-4、IL-10水平在炎性反应升高后向正常方向回落(P<0.001,P<0.05)。电针单穴组、电针双穴组和手针单穴组之间各指标的差异无统计学意义(P>0.05)。结论:三种针刺方法即电针单穴、电针双穴和手针单穴均可提高慢性佐剂性关节炎大鼠的抗炎修复功能,且三种针刺方法治疗效应相当,其机制可能是通过对机体炎性细胞因子的良性诱导,改变相关炎性细胞因子的合成与释放。     

水溶性蜂胶联合阿司匹林对大鼠炎性痛作用

目的探讨水溶性蜂胶联合阿司匹林对大鼠炎性痛作用效应及其可能机制。方法将弗氏完全佐剂造模成功的36只炎性痛大鼠随机均分成6组,炎性痛模型组(模型组)和5个处理组(水溶性蜂胶组,阿司匹林组,水溶性蜂胶+阿司匹林组,水溶性蜂胶+纳洛酮组,水溶性蜂胶+N-硝基-L-精氨酸甲酯(L-NAME)组。随机选取6只大鼠为正常对照组(对照组)。测定各组大鼠的热痛阈潜伏期和机械痛阈值,血清中IL-6、TNF-α及NO的水平,致炎足关节渗出液中PLA2和PGE2的水平。结果与对照组相比,模型组大鼠第1天开始热痛阈潜伏期和机械痛阈值明显降低,热痛觉和机械痛觉过敏持续14 d。模型组大鼠血清中IL-6、TNF-α及NO和渗出液中PLA2和PGE2的水平明显增高。与模型组相比,水溶性蜂胶组、阿司匹林组和水溶性蜂胶+阿司匹林组大鼠热痛阈潜伏期和机械痛阈值均升高,血清中IL-6、TNF-α及NO和渗出液中PLA2和PGE2的水平明显下降。与水溶性蜂胶组相比,水溶性蜂胶+阿司匹林组和水溶性蜂胶+非选择性一氧化氮合酶抑制剂组升高大鼠热痛阈潜伏期和机械痛阈值,而水溶性蜂+纳洛酮组大鼠却明显降低,差异有显著性(P0.05,P0.01)。结论水溶性蜂胶联合阿司匹林对炎性痛大鼠的抗炎镇痛机制可能是抑制细胞因子及炎性介质的产生而实现的,阿片受体途径可能也发挥一定的作用。     

八角麻对大鼠佐剂性关节炎及小鼠免疫功能的影响

 目的:研究八角麻对大鼠佐剂性关节炎及小鼠免疫功能的影响?方法:大鼠佐剂性关节炎模型制作采用弗氏完全佐剂皮下注射(sc)大鼠左后足跖0.1ml?通过研究小鼠末梢血白细胞数、脾脏系数、全血白细胞和腹腔巨噬细胞吞噬功能以及血清溶血素的生成来了解八角麻对小鼠免疫功能的影响?结果:八角麻5g·kg^-1及2.5g·kg^-1给佐剂性关节炎大鼠ig给药11d,每天1次,可明显减轻大鼠佐剂性关节炎;小鼠ig八角麻5g·kg^-1和2.5g·kg^-1×8d,可明显降低末梢血白细胞数和脾脏系数;小鼠ig八角麻5g·kg^-1×7d,可明显降低全血白细胞吞噬功能;小鼠ig八角麻5g·kg^-1×10d,可明显抑制血清溶血素的生成?结论:八角麻对大鼠佐剂性关节炎有明显治疗作用,对小鼠免疫反应有抑制作用?     

豨莶草生品和炮制品抗炎、抗风湿作用的实验研究

目的 :探讨豨莶草生品和炮制品抗炎、抗风湿作用的特点。方法 :采用角叉菜胶诱发大鼠足肿胀 ;二甲苯诱发小鼠耳肿胀 ;大鼠慢性肉芽肿 ;大鼠佐剂性关节炎。结果 :莶草生品和炮制品 6.0 ,2.0g·kg^-1可显著降低角叉菜胶诱发的足肿胀 ,足肿胀抑制率大于 4 0 % ,生品好于炮制品 ;生品和炮制品 6 .0 ,2 .0g·kg^-1可预防大鼠佐剂性关节炎的原发病变和继发病变 ,炮制品在起效时间、作用强度、维持时间等方面明显好于生品。结论 :莶草炮制品对佐剂性关节炎炎症具有明显的抑制作用 ,明显好于生品 ;炮制品和生品对角叉菜胶引起的炎症具有明显的抑制作用 ,两者无明显的差异。     

布洛芬与双氢青蒿素配伍治疗 大鼠佐剂性关节炎的药效及机制初探

目的:研究布洛芬与双氢青蒿素配伍对类风湿关节炎的治疗效果,初步探讨其配伍机制。方法:选择体重160～180 g的SD大鼠随机分为7组:正常对照组、模型对照组、甲氨蝶呤对照组、配伍高剂量治疗组、配伍低剂量治疗组、布洛芬治疗组、双氢青蒿素治疗组。采用佐剂性关节炎(adjuvant arthritis,AA)动物模型,取踝关节进行病理学观察,并运用流式细胞术检测外周血T淋巴细胞亚群、酶联免疫法检测血清白介素-4(IL-4),干扰素-γ(IFN-γ)的水平。结果:配伍组能显著下调外周血CD 4+T细胞百分比和CD 4+T/CD 8+T比值(P<0.01),下调血清INF-γ水平(P<0.01)、上调IL-4水平(P<0.01);与布洛芬组比较,双氢青蒿素组上调IL-4有明显优势(P<0.01),配伍组比单用药物能更有效延缓模型鼠的关节病理损伤。结论:布洛芬与双氢青蒿素配伍能抑制CD 4+T增殖及促进Th1/Th2向右漂移并形成互补,进而增强关节炎治疗效果。     

四妙丸对大鼠佐剂性关节炎作用机制的研究

目的:研究四妙丸对大鼠佐剂性关节炎(adjuvant arthritis,AA)的作用并初步探讨其作用机制,为类风湿关节炎的临床治疗提供实验依据。方法:Wistar大鼠,雄性,随机分为空白对照组(生理盐水)、模型对照组(生理盐水)、雷公藤多苷组(GTT9.45 mg.kg-1)、四妙丸低、中、高剂量组(1 080,2 160,4 320 mg.kg-1)。除空白组外,其余各组采用弗氏完全佐剂注射于大鼠右后足足跖皮内,建立佐剂性关节炎模型;空白组注射等量生理盐水,并分别给予灌胃治疗,观察各组足肿胀值,测定血清中SOD的含量,脾指数和胸腺指数,用sqRT-PCR法检测大鼠滑膜组织中细胞因子IL-1β,IL-6,TNF-αmRNA的表达。结果:四妙丸给药后,大鼠足肿胀均显著减轻,大鼠血清中SOD活性均显著提高,免疫器官脾脏肿大有所缓解,胸腺损害均显著减轻,滑膜组织中IL-1β,IL-6,TNF-αmRNA表达显著降低。结论:四妙丸对大鼠佐剂性关节炎有一定的治疗作用,可减轻炎症反应和关节肿胀,改善关节功能,对类风湿关节炎的临床治疗提供了可靠依据。     

Pharmacological studies on Indian black tea (leaf variety) in acute and chronic inflammatory conditions

Infusions of Indian black tea (BTI), when administered orally, produced significant inhibition of rat paw oedema, induced with carrageenin (pre and post treatment) and arachidonic acid. BTI was also found to inhibit peritoneal capillary permeability and caused a marked reduction of lipopolysaccharide induced PGE(2) generation. In these models, the observed antioedema effect was similar to that of BW755C (a dual inhibitor of cyclooxygenase and 5-lipoxygenase enzymes). BTI was found to scavenge superoxide and hydroxyl radicals, and also protected rat erythrocytes from the damaging effects of hydrogen peroxide. In chronic studies, BTI inhibited granuloma formation along with the reduction of both lipid peroxidation and hydroxyproline content (in the granuloma tissue). Significant antiarthritic activity was observed with regular administration of BTI in the Freund's adjuvant induced model of arthritis. Chronic treatment with BTI (in arthritic rats) resulted in a decrease of paw diameter and tissue lipid peroxidation, along with a restoration of GSH, catalase and superoxide dismutase levels.     

黄芪糖蛋白对佐剂性关节炎大鼠外周血细胞因子及关节滑膜组织形态学的影响

目的探讨黄芪糖蛋白（HQGP）对大鼠佐剂性关节炎（adjuvant arthritis,AA）的治疗作用。方法将雄性Wistar大鼠随机分为正常组、模型组、雷公藤甲素组及HQGP低、中、高剂量组。于右后足跖皮内注射弗氏完全佐剂（CFA）建立大鼠AA模型,1周后给予相应的药物干预;2周后观察致炎侧踝关节肿胀程度以及膝关节滑膜组织形态学的影响,并检测各组大鼠外周血血清细胞因子γ-干扰素（IFN-γ）的水平。结果在治疗两周后,雷公藤甲素组与HQGP中、高剂量组大鼠致炎侧踝关节肿胀以及膝关节滑膜组织增生与模型组相比均有明显改善。HQGP高剂量组大鼠外周血血清IFN-γ水平显著升高。结论HQGP在腹腔注射剂量为3.5mg/kg和7.0mg/kg时,均对AA大鼠的关节炎症有显著改善作用;高剂量HQGP对AA大鼠外周血血清IFN-γ水平有升高作用,这说明在改善AA关节炎症的同时,HQGP还可能提高AA大鼠对抗其他感染的能力。