蜂胶黄酮对卡介苗联合脂多糖诱导小鼠免疫性肝损伤的保护作用

目的 探讨蜂胶黄酮(PF)对卡介苗(BCG)与脂多糖(LPS)诱导小鼠免疫性肝损伤的作用.方法 除正常对照组外,采用BCG和LPS诱导小鼠建立免疫性肝损伤模型,随机分为5组,药物实验组分别用300 ms/ks、150 mg/ks、75 mg/ks的蜂胶黄酮干预后,计算肝指数;检测血清中谷丙转氨酶(ALT)和谷草转氨酶(...     

蜂胶水提物对血管内皮细胞黏附分子表达的影响

目的:采用TNF-α诱导体外培养的人脐静脉内皮细胞(HUCECs)活化,观察蜂胶水提物(WEP)对血管内皮细胞黏附分子表达的影响,从而探讨蜂胶抗动脉粥样硬化的作用及其机制。方法:用50μg/L TNF-α诱导体外培养HUVECs损伤,用50 mg/L、100 mg/L、200 mg/L WEP分别进行干预6 h、12 h、24 h,利用流式细胞仪检测HUVEC表面ICAM-1和VCAM-1表达。结果:与对照组比较,模型组ICAM-1和VCAM-1荧光强度明显升高(P0.01)。与模型组比较,100 mg/L WEP组和200 mg/L WEP组ICAM-1和VCAM-1荧光强度明显降低(P0.01)。不同浓度WEP组ICAM-1和VCAM-1荧光强度随WEP浓度的增加下调。析因分析结果显示,用200 mg/L WEP和氟伐他汀钠(FS)联合预处理组与单一药物预处理组比较,ICAM-1和VCAM-1活性明显降低(P0.01)。结论:WEP能够减少ICAM-1和VCAM-1的表达,且在一定的范围内,有随WEP浓度升高和作用时间延长效应增强的趋势。WEP与FS联合用药,对抑制ICAM-1和VCAM-1表达有协同作用。     

Synergistic Effects of Honey and Propolis toward Drug Multi-Resistant Staphylococcus Aureus, Escherichia Coli and Candida Albicans Isolates in Single and Polymicrobial Cultures

Background: Propolis and honey are natural bee products with wide range of biological and medicinal properties. The study investigated antimicrobial activity of ethyl alcohol extraction of propolis collected from Saudi Arabia (EEPS) and from Egypt (EEPE), and their synergistic effect when used with honey. Single and polymicrobial cultures of antibiotic resistant human pathogens were tested.Material and methods; Staphylococcus aureus (S. aureus),), Escherichia coli (E. coli) and Candida albicans (C.albicans) were cultured in 10-100% (v/v) honey diluted in broth, or 0.08-1.0% (weight/volume) EEPS and EEPE diluted in broth. Four types of polymicrobial cultures were prepared by culturing the isolates with each other in broth (control) and broth containing various concentrations of honey or propolis. Microbial growth was assessed on solid plate media after 24 h incubation.Results; EEPS and EEPE inhibited antibiotic resistant E.coli, and S.aureus, and C.albicans in single and polymicrobial cultures. S.aureus became more susceptible when it was cultured with E.coli or C.albicans or when all cultured together. C.albicans became more susceptible when it was cultured with S.aureus or with E.coli and S. aureus together. The presence of ethyl alcohol or honey potentiated antimicrobial effect of propolis toward entire microbes tested in single or polymicrobial cultures. EEPS had lower MIC toward E.coli and C.albicans than EEPE. When propolis was mixed with honey, EEPS showed lower MIC than EEPE. In addition, honey showed lower MIC toward entire microbes when mixed with EEPS than when it was mixed with EEPE.Conclusion; 1) propolis prevents the growth of the microorganisms in single and mixed microbial cultures, and has synergistic effect when used with honey or ethyl alcohol, 2) the antimicrobial property of propolis varies with geographical origin, and 3) this study will pave the way to isolate active ingredients from honey and propolis to be further tested individually or in combination against human resistant infections.     

蜂胶抑制白血病K562细胞增殖机制的初步研究

目的: 探讨蜂胶对K562细胞增殖、凋亡的影响及其作用机制。方法: 体外培养K562细胞,将不同浓度的蜂胶掺入细胞,采用MTT法检测细胞增殖抑制率,流式细胞术(FCM)检测细胞凋亡率,RT-PCR检测细胞中Nup98 mRNA表达水平。Western blotting检测细胞中Nup98蛋白表达水平。结果: 2 mg/L、20 mg/L和200 mg/L组蜂胶K562细胞的增殖抑制率和凋亡率明显高于对照组,并具有一定的时间和剂量依赖性。高浓度蜂胶作用后,Nup98 mRNA及蛋白表达均明显低于对照组。结论: 蜂胶可抑制K562细胞的增殖,诱导其凋亡,其机制可能与下调Nup98的表达有关。     

在流动血液中蜂胶对血小板活性的抑制作用

目的： 探讨蜂胶提取液对流动状态下血小板活性的影响。方法: 采用体外灌注法，用纤维蛋白原和人Ⅲ型胶原蛋白包被盖玻片，复制血管内膜创伤模型，正常血液流经纤维蛋白原和胶原膜表面后，在切应力为 1 000/s 时，测量血小板粘附率。浓度为0.1 g/L的24%乙醇蜂胶提取液作实验组，加入24%的乙醇作阴性对照组，同浓度的阿魏酸乙醇溶液作阳性对照组。结果: 分别以纤维蛋白原和胶原蛋白膜模拟创伤表面，两实验组血小板粘附率均明显低于阴性对照组，P<0.01；而实验组与阳性对照组比较，均无明显差异，P>0.05。当血液标本流经两个不同的蛋白膜表面时，蜂胶提取液对血小板在纤维蛋白原膜表面粘附率的抑制作用更明显，P<0.01。结论： 蜂胶乙醇提取液能抑制血小板的活化，降低其在纤维蛋白原和胶原蛋白膜表面的粘附活性，对血小板与纤维蛋白原的作用更明显。     

蜂胶水提液对体外培养的血管内皮细胞凋亡的影响

目的：探讨泰山蜂胶水提液对一定浓度的肿瘤坏死因子-α诱导的人脐静脉内皮细胞凋亡的作用。方法:用消化灌注法收集人脐静脉内皮细胞进行体外原代培养，用终浓度为50 μg/L的肿瘤坏死因子-α诱导其凋亡，培养液中分别加入终浓度为50 mg/L、100 mg/L和200 mg/L的蜂胶水提液干预，共同孵育24h，用流式细胞仪和缺口末端标记技术TUNEL检测细胞凋亡率。结果:模型组内皮细胞凋亡率均明显高于对照组，不同浓度蜂胶组内皮细胞凋亡率明显低于模型组（P＜0.01）。结论:泰山蜂胶水提液能抑制肿瘤坏死因子-α诱导的人脐静脉内皮细胞的凋亡，从而起到保护血管内皮细胞的作用。     

The ameliorative effect of propolis against methoxychlor induced ovarian toxicity in rat

A study was designed to evaluate ameliorative effect of propolis against methoxychlor (MXC) induced ovarian toxicity in rat. The organochlorine pesticide (MXC) is a known endocrine disruptor with estrogenic, anti-estrogenic, and anti-androgenic properties. To investigate whether chronic exposure to MXC could cause ovarian dysfunction, two groups of Sprague–Dawley adult female rats were exposed to MXC alone in a dose of 200 mg/kg, twice/weekly, orally or MXC dose as previous plus propolis in a dose of 200 mg/l/day, in drinking water for 10 months. Another two groups of rat were given corn oil (control) or propolis. Multiple reproductive parameters, ovarian weight, serum hormone levels, ovarian oxidative status and ovarian morphology were examined. In MXC-exposed group, there is a significant decrease in body and ovarian weight vs. control. MXC decreases serum estradiol and progesterone levels. A significant increase in the levels of lipid peroxidation was obtained while a significant decrease of the total antioxidant was recorded. Ovarian histopathology showed primary, secondary and vesicular follicles displaying an atretic morphology. Increase in the ovarian surface epithelium height accompanied with vacuolated, pyknotic oocytes were obtained. The previous toxic effects were neutralized by the administration of propolis in MXC + propolis group. The present results suggest that propolis may be effective in decreasing of MXC-induced ovarian toxicity in rat.     

Effect of Brazilian propolis on sneezing and nasal rubbing in experimental allergic rhinitis of mice

We studied the effect of Brazilian propolis on sneezing and nasal rubbing in experimental allergic rhinitis of mice. A single administration of propolis caused no significant effect on both antigen-induced nasal rubbing and sneezing at a dose of 1000?mg/kg, but a significant inhibition was observed after repeated administration for 2 weeks at this dose. Propolis caused no significant inhibitory effect on the production of total IgE level after repeated administration of 1000?mg/kg. The drug also caused no significant inhibition of histamine-induced nasal rubbing and sneezing at a dose of 1000?mg/kg. On the other hand, propolis significantly inhibited histamine release from rat mast cells induced by antigen and compound 48/80 at a concentration of more than 10 μg/ml. These results clearly demonstrated that propolis may be effective in the relief of symptoms of allergic rhinitis through inhibition of histamine release.     

Efficacy of fish liver oil and propolis as neuroprotective agents in pilocarpine epileptic rats treated with valproate

Objective: To evaluate the action of fish liver oil and propolis in pilocarpine epileptic rats treated with the anticonvulsant drug valproate. Methods: Seven groups of rats were treated daily for six months: control; fish liver oil (0.4 ml/kg b.w); propolis (50 mg/kg b.w); pilocarpine-treated rats (epileptic control); epileptic rats treated with valproate (400 mg/kg b.w); groups 6 and 7, epileptic rats treated with valproate plus fish liver oil or propolis. Results: Pilocarpine administration caused a significant increase in hippocampal dopamine and serotonin levels accompanied with a significant decrease in their levels in serum. Lipid peroxidation level and LDH activity in hippocampus were significantly increased after pilocarpine treatment whereas Na⁺/K⁺-ATPase activity and total antioxidant capacity were significantly decreased compared to the controls. Animals treated with the combined treatments showed a significant improvement in tested parameters towards the normal values of the control. Conclusion: Fish liver oil and propolis when given in combination with valproate, neuroprotected against the neurophysiological disorders induced by pilocarpine epilepsy in rats.     

Propolis from Turkey induces apoptosis through activating caspases in human breast carcinoma cell lines

Propolis is a sticky substance that is collected from plants by honeybees that has anti-mutagenic and anti-carcinogenic properties with biological and therapeutic effects. The target of this study was to investigate the anti-apoptotic effect of propolis extracts (PE) on the caspase pathway in the human breast cell line MCF-7 in culture. Seven different propolis extracts, numbered PE 1–7, produced in their natural ecological environment, were collected from the Hacettepe University Beytepe Campus area in Ankara, Turkey. Individual extracts at 0.5, 0.25, 0.125 and 0.063 mg/ml were incubated with MCF-7 cells during 2 days culture. Cell growth and cytotoxicity were measured colorimetrically by MTT assay. Apoptotic cell death was determined by the TUNEL method (terminal deoxynucleotidyltransferase-biotin nick end-labelling) and caspase activity was investigated by immunocytochemistry using antibodies directed against caspase 6, caspase 8 and caspase 9. The results showed that the PE 5 and 6 extracts at 0.125 mg/ml dilution induced apoptosis in association with increased number of TUNEL positive cells. MTT results showed that cultures exposed to the same extracts and at the same dilution experienced better cell growth compared to those cultures exposed to the other extracts. Immunpositivity for all caspases was detected after treatment with all the extracts and at all dilutions, with stronger immunoreactivity for caspase 6 than caspases 8 and 9. Caspase 6 labelling was especially strong in PE 5 and PE 6. We conclude that propolis may have anti-tumour effects by increasing apoptosis through the caspase pathway. Such propolis extracts may be important economically and allow development of a relatively inexpensive cancer treatment.     

Biological evaluation of hesperidin for direct pulp capping in dogs’ teeth

This study compared the biological effect of Hesperidin, Mineral Trioxide Aggregate (MTA)‐Angelus and calcium hydroxide for direct pulp capping. A total of 126 dogs^, teeth were divided according to the post‐treatment evaluation period into three groups (42 teeth each), group I: 2 weeks, group II: 4 weeks and group III: 8 weeks. Each group was further subdivided according to the pulp capping material into three subgroups (14 teeth each), subgroup A (Hesperidin), subgroup B (MTA‐Angelus) and subgroup C (Dycal). Both inflammatory response and dentine bridge formation were assessed by histopathology. All data were statistically analysed. Resolution of the inflammation was recorded by the time with a significant difference between subgroups   within the same group (P<.05). Hesperidin, MTA‐Angelus and Dycal showed either mild or moderate inflammation at 2 weeks with significant differences between subgroups (P < .05). At 4 and 8 weeks, there were no significant differences between subgroups (P > .05). Absence of complete or partial calcified bridge with no odontoblastic layer was reported in all subgroups at 2 weeks while at 4 weeks, the majority of samples in Hesperidin and MTA subgroups showed amorphous calcified deposit. At 8 weeks, there was no significant difference (P > .05) between subgroups except that 78.5% and 92.9% of Hesperidin and MTA‐Angelus samples, respectively, showed moderate dentine bridge. Also, 78.5% of Hesperidin and Dycal samples revealed moderately thick dentine bridge while 78.7% of MTA‐Angelus showed a thin dentine bridge with a significant difference between them (P < .05). In conclusion, Hesperidin is a promising pulp capping material inducing mild inflammation and good dentine bridge formation.     

Effect of topical application of Brazilian propolis on scratching behavior induced by compound 48/80 in mice

Background and aim: We investigated the effect of topical application of Brazilian propolis on scratching behavior induced by compound 48/80 in mice.

Results: Propolis inhibited compound 48/80-induced scratching behavior when applied immediately after treatment with propolis at a dose of 3?mg/site. Dibucaine 0.3?mg/site also significantly inhibited compound 48/80-induced scratching behavior immediately after application. On the other hand, propolis inhibited compound 48/80-induced scratching behavior even 15, 30 and 60?min after application; however, dibucaine showed no significant inhibition of compound 48/80-induced scratching behavior 15, 30 and 60?min after application. In addition, propolis had no effect on increased vascular permeability just after application, but the drug had a significant effect 15, 30 and 60?min after application. On the contrary, histamine-induced scratching behavior was inhibited significantly by propolis just after application. On the other hand, propolis significantly inhibited histamine release from rat mast cells induced by compound 48/80 at a concentration of more than 10?μg/ml.

Conclusion: From these results, it can be concluded that inhibition of scratching behavior induced by topical application occurred by both its local anesthetic and systemic action through inhibition of histamine release.     

Restoration of Glutamine Synthetase Activity,Nitric Oxide Levels and Amelioration of Oxidative Stress by Propolis in Kainic Acid Mediated Excitotoxicity

Background

Propolis has been proposed to be protective on neurodegenerative disorders. To understand the neuroprotective effects of honeybee propolis, glutamine synthetase (GS) activity, nitric oxide (NO), thiobarbituric acid reactive substances (TBARS) and total antioxidant status (TAS) were studied in different brain regions-cerebral cortex (CC), cerebellum (CB) and brain stem (BS) of rats supplemented with propolis and subjected to kainic acid (KA) mediated excitotoxicity.

Materials and Methods

Male Sprague-Dawley rats were divided into four groups; Control group and KA group received vehicle and saline. Propolis group and propolis + KA group were orally administered with propolis (150mg/kg body weight), five times every 12 hours. KA group and propolis + KA group were injected subcutaneously with kainic acid (15mg/kg body weight) and were sacrificed after 2 hrs and CC, CB and BS were separated homogenized and used for estimation of GS activity, NO, TBARS, and TAS concentrations by colorimetric methods. Results were analyzed by one-way ANOVA, reported as mean + SD from 6 animals, and p<0.05 considered statistically significant.

Results

NO was increased (p< 0.001) and GS activity was decreased (p< 0.001) in KA treated group compared to control group as well as propolis + KA treated group. TBARS was decreased and TAS was increased (p< 0.001) in propolis + KA treated group compared KA treated group.

Conclusion

This study clearly demonstrated the restoration of GS activity, NO levels and decreased oxidative stress by propolis in kainic acid mediated excitotoxicity. Hence the propolis can be a possible potential candidate (protective agent) against excitotoxicity and neurodegenerative disorders.     

蜂胶对脂多糖诱导的血管内皮细胞中PC-PLC活性和TLR4表达的影响

目的: 探讨在脂多糖(LPS)诱导的条件下中国蜂胶对血管内皮细胞(VECs)磷脂酰胆碱特异性磷脂酶C (PC-PLC)活性和TLR4表达的影响。方法: 将100 μg/L LPS 加入到含0.5%血清的培养液中培养VECs,经12.5 mg/L的中国蜂胶分别处理12 h和24 h后,SRB法测定细胞存活率;化学法测定NO含量;以L-α-卵磷脂为底物测定PC-PLC的活性;Western blotting检测TLR4、核因子κB p65(NF-κB p65)和p53的表达;细胞内活性氧(ROS)和线粒体膜电位分别通过荧光探针DCHF和JC-1检测。结果: 中国蜂胶处理LPS诱导的血管内皮细胞24 h并不影响细胞存活率,但降低NO的含量和ROS的水平;处理12 h后降低PC-PLC活性和NF-κB p65表达;处理12 h和24 h后降低TLR4和p53的表达。此外,中国蜂胶不影响细胞内线粒体膜电位的水平。结论: 中国蜂胶通过降低PC-PLC活性和TLR4表达,抑制其下游信号分子NF-κB p65、p53的表达和ROS的水平,以及抑制NO的释放,从而发挥抗炎功效。     

Aspergillus fumigatus conidia stimulate lung epithelial cells (TC-1 JHU-1) to produce IL-12, IFNγ, IL-13 and IL-17 cytokines: Modulatory effect of propolis extract

Aspergillus fumigatus conidia are the most prevalent indoors fungal allergens. The interaction between Aspergillus antigens and lung epithelial cells (LECs) result in innate immune functions. The association between Aspergillus conidia and allergic reactions, like allergic bronchopulmonary aspergillosis (ABPA) and asthma have been repeatedly reported. Since conventional therapies for allergy and asthma are limited, finding new promising treatments are inevitable. This study was designed to evaluate the effect of A. fumigatus conidia on IL-12, IFNγ, IL-13 and IL-17 release from mouse LECs and to investigate the effect of propolis on cytokines modulation. Cells were divided to two groups, one was exposed to 3 × 10⁴ conidia of Aspergillus fumigatus and another group was treated by propolis (25 μg/mL) as well as exposed to A. fumigatus conidia. Cytokines IL-13, IL-12, IFNγ and IL-17 were measured at times 0, 6 and 12 hours after exposure using ELISA assay. The results indicated that A. fumigatus could increase the release of the cytokines with IL-13 and IL-17 being the most affected ones whilst treatment with propolis decreased the effects of A. fumigatus on IL-13 and IL-17 production. The results showed that propolis has down regulatory effects on Th2 cytokine, IL-13, and IL-17 production, whereas it caused a significant induction of IL-12, as an important Th1 cytokines by LECs. With respect to the obtained results, propolis extract might be contributed to decrease Th2 responses in allergic asthma phenomenon. However more investigations must be done in future to fully understand its efficacy.     

蜂胶对急性肺损伤大鼠肺组织中NF-κB活性及CD54表达的影响

目的：观察蜂胶对急性肺损伤(ALI)肺组织中细胞间粘附分子—1(CD54)表达和NF—κBp65活化的影响。方法：40只Wistar大鼠分为正常组、肺损伤组、地塞米松组、蜂胶水提组和蜂胶醇提组，脂多糖加油酸两次打击复制ALI模型，观察ALI病理形态，分别用SABC法和SP法测定肺组织中CD54表达和NF—κB p65活性。结果：蜂胶水提液和醇提液均能缓解中性粒细胞的升高，明显减轻肺炎性病变的程度，抑制CD34的表达及NF—κBp65的活化。结论：在ALI肺组织中CD54表达增强和NF—κB p65的活化参与了ALI的发病过程，蜂胶可抑制CD54表达和NF—κB p65的活化，从而减轻肺组织损伤程度。