大鼠真菌性腹腔炎症渗出液的上清液抗真菌作用研究

目的:观察大鼠真菌性腹腔炎症渗出液的上清液中的抗菌物质对真菌的杀菌作用。方法:给大鼠ip活的白色念珠菌Candida albicans造成真菌感染模型,观察腹腔炎症渗出液的上清液对C.albicans、光滑念珠菌C.glabrata、热带念珠菌C.tropicalis的杀菌作用,观察温度对上清液抗真菌作用的影响,并在显微镜下观察杀灭C.albicans的过程。结果:造模后24 h的腹腔炎症渗出大量渗出液,渗出液上清液中含有大量的蛋白质,渗出液上清液对C.albicans,C.glabrata,C.tropicalis具有强的杀菌作用,杀菌作用受温度的影响,56℃处理后杀菌活性几乎丧失,上清液可使C.albicans菌体萎缩,然后胞体破碎死亡。结论:大鼠真菌性腹腔炎症渗出液上清液中存在强的抗真菌物质,具有强的抗真菌作用,可能通过对真菌的细胞壁/细胞膜而发挥作用。其可能是未被关注或未发现的抗菌蛋白/肽。     

牡丹皮等10种中药对白色念珠菌浮游菌 和生物膜作用的研究

目的:观察牡丹皮等10种中药和土槿皮乙酸体外对白色念珠菌浮游菌和生物膜的作用。方法:10种中药为牡丹皮,黄连,丁香,肉桂,高良姜,桂枝,知母,苦参,蛇床子,白花蛇舌草,用水煎煮后浓缩,提取物用DMSO溶解,培养基稀释,体外微量稀释法检测药物对白色念珠菌浮游菌最低抑菌浓度(MIC)和抑制生物膜50%的浓度(SMIC50)的作用。结果:土槿皮乙酸对白色念珠菌的MIC为15.6 mg.L-1,对生物膜的SMIC50是31.2 mg.L-1。牡丹皮和高良姜对白色念珠菌生物膜有一定的抑制作用,SMIC50分别125,250 mg.L-1。黄连,丁香,肉桂,桂枝,知母,苦参,蛇床子,白花蛇舌草在2 000 mg.L-1均没有表现出抑菌活性。结论:体外微量稀释法检测到土槿皮乙酸对白色念珠菌浮游菌和生物膜具有明显的抑制作用,10种中药材中仅牡丹皮和高良姜的水煎液对白色念珠菌生物膜有一定的抑制作用,其他8种中药材的水煎液对白色念珠菌浮游菌和生物膜的抑制作用不明显。对中药抗真菌活性的评价值得进一步探讨。     

黄芩苷对白色念珠菌酶活性的影响

目的运用分光光度法检测黄芩苷作用白色念珠菌后,其琥珀酸脱氢酶(SDH)、细胞色素C氧化酶(CCO)、Ca2 -Mg2 -ATPase活性的改变,以探讨黄芩苷抗白色念珠菌作用靶位,为探索中药活性成分抗真菌作用机理提供依据。方法收集经黄芩苷作用了相应时间的白色念珠菌,制备菌悬液,反复冻融研磨,差速离心法提取白色念珠菌线粒体;Lowry法检测线粒体蛋白质含量;分光光度法检测SDH酶、CCO酶、Ca2 -Mg2 -ATPase活力。结果①0.25,0.5,1mg/ml黄芩苷浓度组白色念珠菌SDH活力明显低于0mg/ml黄芩苷浓度组(P<0.05),且出现黄芩苷浓度越高SDH活力越低的趋势。不同作用时间组间白色念珠菌SDH活力差异无统计学意义(P>0.05)。②白色念珠菌经不同浓度黄芩苷作用不同时间后,其CCO酶活力均无明显差异(P>0.05)。③0.25,0.5,1mg/ml黄芩苷浓度组与0mg/ml黄芩苷浓度组比较,Ca2 -Mg2 -ATPase活力差异显著,黄芩苷浓度越高,Ca2 -Mg2 -ATPase活力越低,作用时间对Ca2 -Mg2 -AT-Pase活力没有明显影响(P>0.05)。结论①黄芩苷具有明显的抗白色念珠菌作用;②黄芩苷可降低白色念珠菌SDH酶活力;③黄芩苷对白色念珠菌CCO酶活力未见明显影响;④黄芩苷可降低白色念珠菌Ca2 -Mg2 -ATPase活力。     

中草药体外抗白色念珠菌的实验研究

探索有效中草药对白色念珠菌抗菌作用。方法：采用菌基混合加药法、双倍稀释法体外测定８种中草药抗白色念珠菌作用的效果。结果：生大蒜汁、七叶一枝花有很强的抗白色念珠菌作用,其ＭＩＣ分别为１．０ｍｇ／ｍｌ和１．５ｍｇ／ｍｌ,抗菌效价为３．１２ｍｇ／ｍｌ和６．２５ｍｇ／ｍｌ;丁香、一口钟其ＭＩＣ均为２．０ｍｇ／ｍｌ,抗菌效价均为１２．５ｍｇ／ｍｌ,表示抗菌作用较强;虎杖、土槿皮、木鳖子抗菌作用较弱,其ＭＩＣ     

真菌安胶囊体外抗白色念珠菌的实验研究

目的探索真菌安胶囊对白色念珠菌抗菌的作用。方法:采用菌基混合加药法、双倍稀释法,体外测定真菌安胶囊抗白色念珠菌作用的效果。结果:真菌安胶囊有明显的抗白色念珠菌作用,其MIC为2.0mg/ml。结论:为真菌安胶囊治疗白色念珠菌感染性疾病,提供了实验依据。     

抗白色念珠菌中草药筛选概况

近年来由于抗生素、皮质类固醇等药物的广泛应用,白色念珠菌病(白念)较为常见,患者有所增加。目前临床应用的抗真菌药物主要为多烯抗生素、氨基甲酸酯及咪唑类等。这些药物疗效肯定,但存在一些毒副反应,理想的药物极少。因而急待开发疗效高,毒副作用小的新型药物。中草药具有悠久的历史,从中开发抗真菌药物已进行大量的筛选工作,并取得一定的进展。本文概述有关抗白念方面的研究结果。     

大蒜素对白色念珠菌毒力因子作用机制的研究

目的探讨大蒜素对白色念珠菌毒力因子的作用机制。方法采用CLSI-M27-A3微量液基稀释法检测大蒜素对白色念珠菌的最小抑菌浓度(MIC);通过时间-杀菌曲线检测大蒜素的杀菌作用;Spider液体培养基中观察大蒜素对白色念珠菌菌丝生长的影响;qRT-PCR法检测大蒜素对白色念珠菌菌丝相关基因表达的影响;卵黄乳液琼脂平板法检测大蒜素对白色念珠菌细胞外磷脂酶活性的影响。结果大蒜素对白色念珠菌的MIC值为12.5～25μg/mL;时间-杀菌曲线显示2 MIC、4 MIC浓度的大蒜素对白色念珠菌生长保持有效的抑制作用;在Spider液体培养基中,1 MIC、2 MIC、4 MIC浓度的大蒜素能明显抑制白色念珠菌菌丝的生长;qRT-PCR结果显示,在1 MIC、2 MIC、4 MIC浓度的大蒜素作用下,白色念珠菌菌丝相关基因表达下调;当大蒜素浓度≥1/2 MIC时,白色念珠菌细胞外磷脂酶的活性降低。结论大蒜素可通过抑制白色念珠菌菌丝生长、调节菌丝相关基因的表达,以及抑制细胞外磷脂酶活性等,发挥抑制白色念珠菌毒力因子的作用。     

妇安泡腾片抗白色念珠菌的体外药敏试验

中草药治疗真菌感染有不少报道。本研究观察本院自制妇安泡腾片,对白色念珠菌引起的阴道炎的抗菌作用。兹将结果报告如下。     

参芪扶正液辅助治疗肺白色念珠菌感染的实验研究

目的探讨参芪扶正液辅助抗真菌药治疗肺白色念珠菌感染小鼠的效果。方法经环磷酰胺诱导小鼠免疫功能下降,并经肺感染白色念珠菌,之后分别给予单独抗真菌药和伍用参芪扶正液治疗,比较各组的疗效。结果参芪扶正液辅助抗真菌药治疗肺白色念珠菌感染较单独使用抗真菌药效果显著,2组比较有显著性差异(P<0.01)。结论参芪扶正液与抗真菌药伍用可提高生存率,增强抗真菌药物的疗效。     

Effects of Ginkgo biloba on haemostatic factors and inflammation in chronic peritoneal dialysis patients

Ginkgo biloba extract decreased plasma D-dimer concentration, a marker of intravascular coagulation, in chronic peritoneal dialysis patients. Blood levels of fibrinogen, von Willebrand factor, hs-CRP, albumin and liver enzyme levels were not significantly changed. No bleeding episode was reported. These results suggest that Ginkgo biloba extract was effective in partially reversing the thrombogenic coagulation profile without increasing the risk of bleeding.     

Effect of resveratrol on NF-kappaB activity in rat peritoneal macrophages

This study was to investigate the inhibitive effect of resveratrol (RESV) on nuclear factor kappa B (NF-kappaB) expression and activity induced by lipopolysaccharide (LPS) in rat peritoneal macrophages (PMA). Male Sprague-Dawley (SD) rats were randomly divided into 7 groups, including control group, LPS group and RESV I-V group. In the LPS group, PMA were incubated in DMEM containing LPS (10 microg/ml), whereas in control group, PMA were incubated in DMEM only. In the RESV I-V groups, PMA were incubated in DMEM containing LPS (10 microg/ml) and different concentrations of RESV. After 24 hours of incubation, NF-kappaB activity in PMA, and the levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1) and nitric oxide (NO) in the culture medium were measured. In the concentrations of 1.25-5 microg/ml, RESV had a dose- dependent inhibitive effect on NF-kappaB activity in PMA as well as the expressions of TNF-alpha, IL-1 and NO in the culture medium contrasted with the LPS group. There was no significant difference in the levels of these pro-inflammatory factors between the groups of 5 microg/ml and 10 microg/ml RESV. In conclusion, RESV has the potential for the future application of preventing inflammatory diseases involving PMA.     

痛风克冲剂对痛风大鼠前列腺素E_2的影响

目的　通过观察痛风克冲剂对大鼠前列腺素 (PG)E2 及其踝关节肿胀程度的改善情况 ,探讨其对痛风的治疗机制。方法　将 60只Wistar大鼠随机均分成空白组、模型组、痛风克组、秋水仙组、别嘌呤醇组、丙磺舒组 6组。造模前 2d开始用不同药剂连续灌胃 ,7d后测各组大鼠PGE2 及其踝关节肿胀程度。结果　痛风克与秋水仙碱组可明显降低大鼠PGE2 及踝关节肿胀程度 (P <0 .0 5) ,且两组疗效相近 (P >0 .0 1 ) ,并优于别嘌呤醇组和丙磺舒组 (P <0 .0 5)。结论　痛风克冲剂可降低炎症递质PGE2 ,从而达到抗炎、消肿的目的     

丹红注射液对高糖引起腹膜间皮细胞损伤的作用

目的研究丹红注射液对高糖所致大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMCs)氧化应激、炎症反应及其损伤的影响。方法原代培养RPMCs,随机分为正常对照组、高糖组(100 mmol/L高糖分别作用RPMCs12、24、48、72 h)、丹红注射液组(80 m L/L丹红注射液单独作用RPMCs 48 h)、高糖+丹红注射液组(100 mmol/L高糖分别与40、80、160 m L/L丹红注射液共同作用48 h)。MTT测定各组RPMCs增值率。酶联免疫法检测细胞培养液中白介素-18(IL-18)、白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)表达,试剂盒检测Caspase-3活性及活性氧(ROS)、丙二醛(MDA)、还原型谷胱甘肽(GSH)、超氧化物岐化酶(SOD)含有量,实时定量RT-PCR法检测各组凋亡相关基因(Bcl-2、Bax)、纤维连接蛋白(FN)、内皮素1(EDN1)和血红素氧化酶1(HMOX1)mRNA的表达。Western blot法检测EDN1和HMOX1蛋白表达。结果高糖明显抑制RPMCs的增殖,增强Caspase-3活性,增加IL-18、IL-6和TNF-α的表达量,诱导RPMCs中ROS、MDA过度产生和抑制GSH、SOD产生,同时增加EDN1 mRNA与蛋白、Bax mRNA、FN mRNA的表达,降低HMOX1 mRNA与蛋白、Bcl-2 mRNA表达。而丹红注射液+高糖组的RPMCs的增殖率明显高于高糖组,Caspase-3活性降低,IL-18、IL-6和TNF-α的表达减少,细胞内ROS水平及上清液中MDA的含有量显著降低,同时EDN1 mRNA与蛋白,Bax、FN mRNA的表达下降,而上清液中SOD及GSH的含有量明显上升,HMOX1 mRNA与蛋白、Bcl-2 mRNA表达增高。结论丹红注射液可减少高糖所致细胞凋亡,抑制高糖所致的氧化应激及炎性反应,进而起到对RPMCs的保护作用。     

生地注射液对脂多糖诱导大鼠肺部炎症的影响

目的：研究生地注射液对大鼠肺部炎症的影响。方法：气道滴入大肠杆菌脂多糖(LPS)建立大鼠肺部炎症模型；随机分正常对照组、模型组、药物组,各组均采用静脉注射给药方式,2次给药,模型组给生理盐水,药物组每次给生地注射液380 mg·kg^-1(以生地注射液中固形物量计),进行支气管肺泡灌洗液(BALF)中细胞分类计数；测定BALF中肿瘤坏死因子(TNF-α),超氧阴离子自由基(O^-₂),中性粒细胞髓过氧化酶(MPO)水平；进行肺组织病理检查。结果：生地注射液静脉给药可明显抑制LPS诱导的白细胞总数和中性粒细胞数目升高,对淋巴单核细胞数目无明显影响；明显抑制TNF-α,O^-2,MPO水平升高；明显抑制LPS诱导的肺组织中性粒细胞浸润、气管黏膜水肿,对肺组织损伤无影响。结论：生地注射液有一定的抗LPS致大鼠肺部炎症的作用,初步推断抗炎、抑制细胞因子释放、抑制炎症介质释放和抗氧化是其治疗慢性阻塞性肺疾病的作用机制之一。     

甘草酸单铵配伍复方丹参注射液对大鼠盆腔粘连及组织生长因子的影响

目的:观察甘草酸单铵盐(MAG, monoammonium glycyrrhetate)配伍复方丹参注射液对大鼠子宫角粘连的预防作用及其作用机理.方法:制备大鼠宫角粘连模型,观察粘连评分,测定粘连组织中血管内皮生长因子(VEGF)、成纤维细胞生长因子(bFGF)的阳性表达率.结果:除复方丹参组外,各用药组均明显降低粘连评分和严重粘连百分率.与MAG组比较,MAG 复方丹参组、Dex组严重粘连百分率更低.与粘连组比较,MAG组、复方丹参组、MAG 复方丹参组、Dex组的VEGF、bFGF阳性细胞百分率均显著降低.结论:MAG、MAG配伍复方丹参可显著减少大鼠盆腔粘连的形成,尤以MAG配伍复方丹参组显著,二者可能具有协同作用,对粘连形成中晚期血管及纤维组织增生具有抑制作用可能是其作用机理之一.     

The anti-Candida activity of Thymbra capitata essential oil: effect upon pre-formed biofilm

Ethopharmacological relevance

Thymbra capitata essential oil is traditionally considered to exhibit powerful antiseptic properties, thus being used to treat cutaneous infections. The aim of the present study was to evaluate the effect of Thymbra capitata essential oil upon pre-formed biofilm of different Candida strains while comparing it with the activity against planktonic cells.

Materials and methods

Fifteen Candida isolates were included, corresponding to clinical and collection type strains. Essential oil was obtained by hydrodistillation and its composition analysed by GC/MS. Activity upon planktonic cells was evaluated according to M27-A3 macromethod. Its effect upon 24 h preformed biofilm biomass was determined using the crystal violet procedure and the metabolic activity was studied applying the XTT/menadione technique.

Results

Biofilm biomass and metabolic activity of all tested species were reduced up to 50% at MIC values. The effect was more pronounced at double MIC values, achieving >80% reduction, except for Candida albicans that presented a more resistant profile (62%).

Conclusion

Thymbra capitata essential oil presented an important effect upon Candida biofilms. It is proposed as a valuable antifungal product to be used in an appropriate pharmaceutical formulation for the management of resistant mucocutaneous candidosis.     

Effect of ganhuangenin obtained from Scutellaria radix on the chemical mediator production of peritoneal exudate cells and immunoglobulin E level of mesenteric lymph node lymphocytes in Sprague-Dawley rats

We previously showed that ganhuangenin (GHG) has beneficial antioxidative properties against lipid peroxidation in tissues of Sprague-Dawley (SD) rats. In this study, the effect of GHG on immunoglobulin E (IgE), histamine and leukotriene B(4) (LTB(4)) levels of cells isolated from SD rats was examined with regard to the manifestation of the type I allergic reaction. We showed that GHG inhibited the IgE production of mesenteric lymph node (MLN) lymphocytes. In the presence or absence of concanavalin A (ConA), the concentration used did not exert a toxic effect against MLN lymphocytes. Interestingly, the increase in the IgE content and lipid peroxidation induced by ConA was alleviated in the presence of GHG. Moreover, GHG also inhibited histamine release from the peritoneal exudate cells stimulated with a calcium ionophore, A23187, or with the histamine releaser, compound 48/80. In the case of LTB(4), GHG markedly inhibited its release at a concentration of 100 microM. Thus, it is concluded that GHG may block the common pathway for the release of histamine and LTB(4), and that the IgE level is responsible for the lipid peroxidation induced by ConA.     

Co-cultured adventitious roots of Echinacea pallida and Echinacea purpurea inhibit lipopolysaccharide-induced inflammation via MAPK pathway in mouse peritoneal macrophages

Objective: In order to elucidate the biological activity of the co-cultured adventitious roots (ARs) of Echinacea pallida and Echinacea purpurea and provide theoretical basis for its application, and the anti-inflammatory activities and potential mechanisms of co-cultured ARs were studied. Methods: The experimental materials were obtained by bioreactor co-culture technology and used in the activity research. In this study, mouse macrophages induced by lipopolysaccharide (LPS) were used as in vitro model. Different concentrations of AR extract (50?400 g/mL) were used to treat cells. The expression of pro-inflammatory cytokines was determined using enzyme linked immunosorbent assay. The inducible nitric oxide synthase and cyclooxygenase-2 expression, mitogen-activated protein kinase (MAPK) phosphorylation, and the inhibitor of nuclear factor-kappa B-α levels were determined by the Western blot analysis. Results: In the co-cultured ARs, total flavonoids and total caffeic acid were determined, and the contents of both bioactive compounds were significantly higher than those ARs from the single-species culture. Compared with the control group, the large amount of pro-inflammatory mediators was released after LPS stimulation. However, in the extract groups with different concentrations (25, 50, and 100 g/mL), the production of these pro-inflammatory mediators was inhibited in a dose-dependent manner. Furthermore, the levels of phosphorylation of MAPK proteins, including p-p38, p-c-Jun N-terminal kinase, and p-extracellular regulated protein kinases were significantly (P < 0.05) decreased in the extract groups, revealing that the AR extract probably involved in regulating the MAPK signaling pathway. Conclusion: Collectively, our findings suggested that the co-cultured ARs of E. pallida and E. purpurea can inhibit production of pro-inflammatory mediators in mouse peritoneal macrophages and possess the anti-inflammatory effect by regulating MAPK signaling pathways.