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1.
选取传统中药提取单体化合物α-倒捻子素和8-甲氧基补骨脂素,通过2种药物敏感性实验方法MABA(Microdilution Alamar Blue Assay)分析与试管法分别测定了两种化合物对结核分枝杆菌H37Rv(ATCC27294)和H37Ra(ATCC25177)的体外抗菌活性,同时还测定了9种阳性抗结核药物对这两种结核菌的最低抑菌浓度(MIC),比较2种方法的检测结果。结果表明:α-倒捻子素对结核分枝杆菌H37Rv(ATCC27294)和H37Ra(ATCC25177)的最低抑菌浓度均为6.25μg/mL,而8-甲氧基补骨脂素对结核分枝杆菌H37Rv(ATCC27294)和H37Ra(ATCC25177)的最低抑菌浓度均为128μg/mL。Alamar B1ue法与试管法相比较,完全符合率是90%(9/11),具有较好的一致性。本研究结果表明Mamar B1ue法是一种快速、简便测定药物对结核分枝杆菌MIC的方法。本研究为α-倒捻子素和8-甲氧基补骨脂素的进一步开发利用和抗分枝杆菌机制研究打下了基础。  相似文献   

2.
MABA法与二倍稀释法测定齐墩果酸体外抗结核菌活性   总被引:1,自引:0,他引:1  
为筛选有开发价值的抗结核中草药,通过儿拉姆兰微板分析法(MABA)与二倍稀释法分别测定齐墩果酸对结核分支杆菌H37Rv(ATCC 27294)和H37Ra(ATCC 25177)的体外抗菌活性,测定最低抑菌浓度(MIC),同时对两种方法的检测结果进行了比较;以噻唑蓝(MTT)法进行了齐墩果酸对Balb/c小鼠的脾细胞和肝细胞的细胞毒性试验。结果表明,齐墩果酸对这两种结核菌的最低抑菌浓度均为16μg/mL,对Balb/c小鼠的脾细胞和肝细胞均无毒性。MABA法与试管法相比较,完全符合率为90%(9/10),具有较好的一致性。本研究结果为齐墩果酸作为抗结核菌药物的开发及其抗菌机制研究奠定了一定的基础。  相似文献   

3.
中药单体化合物光甘草定的体外抗菌活性研究   总被引:1,自引:0,他引:1  
选取传统中药甘草提取单体化合物光甘草定,通过MABA药物敏感性实验方法研究其对结核分枝杆菌H37Rv(ATCC27294)和H37Ra(ATCC25177)的体外抗菌活性,并通过NCCLS标准方法测定了光甘草定对金黄色葡萄球菌ATCC25923和ATCC29213、粪肠球菌ATCC29212和大肠杆菌ATCC25922的最低抑菌浓度(MIC)。结果表明:单体化合物光甘草定对结核分枝杆菌H37Rv(ATCC27294)和H37Ra(ATCC25177)有很好的抗菌活性,最低抑菌浓度均为25μg/m;光甘草定对实验中检测的金黄色葡萄球菌、粪肠球菌和大肠杆菌也有抗菌活性,并且对革兰氏阳性杆菌的抗菌活性高于革兰氏阴性杆菌。本研究为光甘草定的进一步开发利用和抗菌机制研究打下了基础。  相似文献   

4.
研究选取中药单体18β-甘草次酸联合一线抗结核药物异烟肼(INH)、利福平(RFP)和链霉素(SM),利用MABA法检测其体外单独及联合作用于牛源结核分枝杆菌的抗菌活性。18β-甘草次酸单独作用于结核分枝杆菌标准株(ATCC 27294)和牛分枝杆菌标准株(ATCC 19210)的最低抑菌浓度(MIC)分别为50和100 μg/mL。临床分离的2株敏感株和6株耐药株的MIC值分别为25~50和100~200 μg/mL。18β-甘草次酸联合INH、RFP和SM作用于6株耐药株均具有协同作用,且MIC值明显下降,INH的MIC值下降2~32倍(FICIs 0.125~0.375),RFP的MIC值下降4~8倍(FICIs 0.240~0.490),SM的MIC值下降4~16倍(FICIs 0.165~0.460)。中药单体药物对正常细胞BHK-21具有较低的细胞毒性,对肝癌细胞SMMC具有较好抑制作用。结果表明,18β-甘草次酸与抗结核药物INH、RFP和SM联合使用对结核分枝杆菌具有较好的抗菌药理活性。  相似文献   

5.
沙冬青种子总黄酮活性成分分析及对小鼠免疫功能的影响   总被引:2,自引:2,他引:0  
本研究旨在通过高效液相色谱-质谱联用(LC-MS)和高效液相色谱(HPLC)技术对沙冬青种子总黄酮主要活性成分进行分析及含量测定,并探索沙冬青种子总黄酮对小鼠免疫功能的影响。采用LC-MS分析沙冬青种子总黄酮主要活性成分,在HPLC优化条件下用标准品对主要活性成分的含量进行测定;采用MTT法和ELISA试剂盒检测沙冬青种子总黄酮对小鼠脾淋巴细胞增殖和IL-2、IL-4生成的影响,以及总黄酮对肝脏KCs细胞的增殖和NO、NOS含量的影响;运用定量溶血分光光度法和血清溶血素HC_(50)测定法检测不同浓度沙冬青种子总黄酮对小鼠抗体生成细胞和血清溶血素的影响。结果显示,沙冬青种子总黄酮中主要活性成分为芒柄花素、7,3’-二羟基-4’-甲氧基黄酮、黄豆黄素和穗花杉双黄酮,其中,芒柄花素含量高达52.48%,其次为7,3’-二羟基-4’-甲氧基黄酮,含量为11.20%。在2.5~100μg/mL总黄酮作用下,沙冬青种子总黄酮对脾淋巴细胞的增殖和细胞因子的产生具有明显的促进作用,尤其在20μg/mL时脾淋巴细胞增殖率和IL-2的分泌量分别高达217.62%和159.661 pg/mL,与空白对照组相比差异极显著(P0.01);在40μg/mL时,脾淋巴细胞IL-4的分泌量高达149.274 pg/mL,极显著高于空白对照组(P0.01)。沙冬青种子总黄酮对KCs细胞的增殖及NO和NOS分泌也具有明显促进作用,在60μg/mL时,与空白对照组相比,KCs细胞增殖率达67.77%,NO和NOS的分泌量分别达180.106和29.942μmol/L(P0.01)。此外,沙冬青种子总黄酮能明显促进小鼠体内抗体生成细胞和血清溶血素含量的增加。综上表明,沙冬青种子总黄酮主要活性成分为芒柄花素和7,3’-二羟基-4’-甲氧基黄酮。沙冬青种子总黄酮对小鼠免疫活性细胞的增殖、相关免疫细胞因子和抗体的产生与释放等都具有显著的促进作用。  相似文献   

6.
抑制鹿源多药耐药结核菌中药的筛选   总被引:1,自引:0,他引:1  
为了筛选对鹿源多药耐药结核菌有效的中药耐药抑制剂,通过MABA法检测24种中药的水和乙醇提取物对多药耐药结核菌及标准菌H37Rv的体外抗菌效果,将筛选出的抗菌效果好的黄连醇提物和黄连水提物采用紫外分光光度法进行活性成分含量测定,同时以MABA法与标准品比较体外抗结核菌活性。结果显示,黄连、独活和侧柏叶有明显的抗结核活性,其中黄连水粗提物和乙醇粗提物效果最佳,测得它们活性成分即总生物碱含量分别为22.74%和35.23%,并显示二者与小檗碱的体外抗结核作用相同。  相似文献   

7.
本研究旨在通过高效液相色谱-质谱联用(LC-MS)和高效液相色谱(HPLC)技术对沙冬青种子总黄酮主要活性成分进行分析及含量测定,并探索沙冬青种子总黄酮对小鼠免疫功能的影响。采用LC-MS分析沙冬青种子总黄酮主要活性成分,在HPLC优化条件下用标准品对主要活性成分的含量进行测定;采用MTT法和ELISA试剂盒检测沙冬青种子总黄酮对小鼠脾淋巴细胞增殖和IL-2、IL-4生成的影响,以及总黄酮对肝脏KCs细胞的增殖和NO、NOS含量的影响;运用定量溶血分光光度法和血清溶血素HC50测定法检测不同浓度沙冬青种子总黄酮对小鼠抗体生成细胞和血清溶血素的影响。结果显示,沙冬青种子总黄酮中主要活性成分为芒柄花素、7,3'-二羟基-4'-甲氧基黄酮、黄豆黄素和穗花杉双黄酮,其中,芒柄花素含量高达52.48%,其次为7,3'-二羟基-4'-甲氧基黄酮,含量为11.20%。在2.5~100 μg/mL总黄酮作用下,沙冬青种子总黄酮对脾淋巴细胞的增殖和细胞因子的产生具有明显的促进作用,尤其在20 μg/mL时脾淋巴细胞增殖率和IL-2的分泌量分别高达217.62%和159.661 pg/mL,与空白对照组相比差异极显著(P<0.01);在40 μg/mL时,脾淋巴细胞IL-4的分泌量高达149.274 pg/mL,极显著高于空白对照组(P<0.01)。沙冬青种子总黄酮对KCs细胞的增殖及NO和NOS分泌也具有明显促进作用,在60 μg/mL时,与空白对照组相比,KCs细胞增殖率达67.77%,NO和NOS的分泌量分别达180.106和29.942 μmol/L(P<0.01)。此外,沙冬青种子总黄酮能明显促进小鼠体内抗体生成细胞和血清溶血素含量的增加。综上表明,沙冬青种子总黄酮主要活性成分为芒柄花素和7,3'-二羟基-4'-甲氧基黄酮。沙冬青种子总黄酮对小鼠免疫活性细胞的增殖、相关免疫细胞因子和抗体的产生与释放等都具有显著的促进作用。  相似文献   

8.
BSN-37是从牛脾脏分离鉴定的含有37个氨基酸的新抗菌肽。为了阐明该抗菌肽的生物活性,本试验检测了其对大肠杆菌(ATCC25922)、鸡白痢沙门菌(BNCC124693)、金黄葡萄球菌(ATCC25923)、白色念珠菌(ATCC90029)及5株临床大肠杆菌和7株临床鼠伤寒沙门菌的最低抑菌浓度(MICs);同时,测定其对兔血红细胞悬液的溶血性和对Vero细胞、小鼠腹腔巨噬细胞(RAW264.7)和鸡成纤维细胞(CEF)的细胞毒性。结果表明,BSN-37对大肠杆菌ATCC25922、鸡白痢沙门菌(BNCC124693)MICs值分别为33.33和25.00mg/L;对所检测的临床耐药大肠杆菌(卡那霉素MICs≥320.00mg/L)和鼠伤寒沙门菌的MICs值均低于50.00mg/L;对金黄葡萄球菌和白色念珠菌的MICs值均大于100.00mg/L。400mg/L的BSN-37与兔血红细胞体外37℃恒温孵育1h后,红细胞溶血率仅为1.34%;50mg/L的BSN-37与RAW264.7细胞作用后,细胞存活率为96.3%,当BSN-37质量浓度为100,200和400mg/L时,ERO、RAW264.7和CEF 3种细胞的存活率均大于100.00%。以上结果证实,BSN-37对革兰阴性肠道菌有较强抑菌活性,溶血性很低且无细胞毒性,具有良好的临床药物开发前景。  相似文献   

9.
为设计研发新型天蚕素类抗菌肽(Cecropin-like antibacterial peptide,CLP),本研究参照抗菌肽Cecropin P1的作用机理和氨基酸组成,通过人工方法设计合成CLP,并分别测定其体外和体内抗菌活性。体外抑菌试验结果显示,新型CLP对金黄色葡萄球菌(ATCC29213)、禽多杀性巴氏杆菌(CVCC474)、鸡白痢沙门氏菌(ATCC10398)、鸡致病性大肠杆菌O1均具有良好的抑菌活性,对这4种细菌的最小抑菌浓度(MIC)分别为6.25μg/mL、1.56μg/mL、3.12μg/mL和6.25μg/mL。酸稳定性试验显示,新型CLP在pH值为6~8时活性最高,热稳定性试验显示新型CLP于100℃加热2 h后仍具有抑菌活性。在对鸡白痢沙门氏菌感染雏鸡的体内疗效试验中,新型CLP显示了良好的治疗效果。本研究为临床上设计有效的天蚕素类抗菌肽提供了参考数据。  相似文献   

10.
【目的】研究异丙氧苯胍与黏菌素联用对多杀性巴氏杆菌的体内外抗菌作用,为开发有效的新型黏菌素增效剂及临床防治多杀性巴氏杆菌病提供数据支持。【方法】通过微量肉汤稀释法测定异丙氧苯胍与黏菌素对多杀性巴氏杆菌的最小抑菌浓度,再通过棋盘法联合药敏试验和时间杀菌曲线评价异丙氧苯胍与黏菌素联用对多杀性巴氏杆菌的体外抗菌作用,并进一步通过建立小鼠肺部感染多杀性巴氏杆菌模型来评价两者联用的体内抗菌效果。【结果】药敏试验结果显示,异丙氧苯胍对多杀性巴氏杆菌无抗菌作用(MIC>256μg/mL),黏菌素对受试菌株的最小抑菌浓度范围为1~8μg/mL。联合药敏试验结果显示,异丙氧苯胍与黏菌素联用时能增强黏菌素的抗菌活性(分级抑菌浓度指数在0.094~0.313),表现出良好的协同抗菌作用。体外时间杀菌曲线结果进一步表明,联合用药组可显著降低细菌数量,当亚抑菌浓度(0.5μg/mL)的黏菌素与异丙氧苯胍联用时即可达到杀菌效果。在小鼠肺部感染模型中,与黏菌素或异丙氧苯胍单药组相比,异丙氧苯胍与黏菌素联合组能显著或极显著降低小鼠肺部多杀性巴氏杆菌的载菌量(P<0.05或P<0.01)。HE染色观察...  相似文献   

11.
The objectives of this study were to determine the plasma and pulmonary disposition of gamithromycin in foals and to investigate the in vitro activity of the drug against Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) and Rhodococcus equi. A single dose of gamithromycin (6 mg/kg of body weight) was administered intramuscularly. Concentrations of gamithromycin in plasma, pulmonary epithelial lining fluid (PELF), bronchoalveolar lavage (BAL) cells, and blood neutrophils were determined using HPLC with tandem mass spectrometry detection. The minimum inhibitory concentration of gamithromycin required for growth inhibition of 90% of R. equi and S. zooepidemicus isolates (MIC(90)) was determined. Additionally, the activity of gamithromycin against intracellular R. equi was measured. Mean peak gamithromycin concentrations were significantly higher in blood neutrophils (8.35±1.77 μg/mL) and BAL cells (8.91±1.65 μg/mL) compared with PELF (2.15±2.78 μg/mL) and plasma (0.33±0.12 μg/mL). Mean terminal half-lives in neutrophils (78.6 h), BAL cells (70.3 h), and PELF (63.6 h) were significantly longer than those in plasma (39.1 h). The MIC(90) for S. zooepidemicus isolates was 0.125 μg/mL. The MIC of gamithromycin for macrolide-resistant R. equi isolates (MIC(90)=128 μg/mL) was significantly higher than that for macrolide-susceptible isolates (1.0 μg/mL). The activity of gamithromycin against intracellular R. equi was similar to that of azithromycin and erythromycin. Intramuscular administration of gamithromycin at a dosage of 6 mg/kg would maintain PELF concentrations above the MIC(90) for S. zooepidemicus and phagocytic cell concentrations above the MIC(90) for R. equi for approximately 7 days.  相似文献   

12.
Rapidly growing mycobacteria (RGM) and Nocardiae can cause severe or refractory infections in cats and dogs. Prolonged antibacterial therapy is required to cure these infections. As fluoroquinolones have been used in combination therapy for treating RGM infections, isolates from the Mycobacterium smegmatis cluster (n=64), Mycobacterium fortuitum cluster (n=17), and M. mageritense cluster (n=2), collected from feline and canine patients, underwent susceptibility testing to pradofloxacin. The MIC(50), MIC(90) and tentative epidemiological cut-off (ECOFF) values as determined by microbroth dilution susceptibility testing that inhibited growth of the M. smegmatis and M. fortuitum clusters were 0.063, 0.125 and ≤ 0.25; and 0.125, 0.250 and ≤ 1.0 μg/mL, respectively. E-Test results showed similar trends but MICs were lower than those for microbroth dilution. In summary, pradofloxacin demonstrated effective in vitro activity against RGM isolates. Additionally, veterinary isolates of Nocardia nova (n=18), Nocardia farcinica (n=3) and Nocardia cyriacigeorgica (n=1) underwent microbroth dilution testing to ciprofloxacin, enrofloxacin and pradofloxacin. The MIC(50) and MIC(90) of pradofloxacin, ciprofloxacin and enrofloxacin that inhibited growth of Nocardia nova isolates were 2 (4), 8 (16), 16 (32) μg/mL, respectively. The tentative ECOFF values for pradofloxacin and ciprofloxacin were 32 μg/mL and for enrofloxacin 64 μg/mL. The MIC or MIC range for the three N. farcinica isolates of pradofloxacin, ciprofloxacin and enrofloxacin were 0.25-0.5, 2 and 2 μg/mL and for the single N. cyriacigeorgica isolate were 1, 4 and 4 μg/mL, respectively. On the basis on these results, fluoroquinolones appear to have limited therapeutic potential for most Nocardia infections.  相似文献   

13.
本研究旨在寻找防治鲟鱼病害的有效中草药药物。用水煎煮法提取天然物中草药有效成分,采用试管法测定各提取物对维氏气单胞菌的最低抑菌浓度,最后通过正交试验确定了4种中草药提取物体外抑菌的最优组方。33种中草药提取物对维氏气单胞菌抑菌效果最好的是五倍子,其MIC为0.97 μg/mL;其次是地榆和黄芩,两者MIC均为1.95 μg/mL;马齿苋和苦参抑菌效果最差,MIC为250 μg/mL。选取五倍子、地榆、地锦草和连翘进行正交试验,其最优组方为1∶2∶4∶8,该组方对维氏气单胞菌有明显的抑制作用,为进一步应用中草药防治水产养殖病害提供依据。  相似文献   

14.
β‐Defensins (BDs) are highly conserved antimicrobial peptides important in innate defence against bacteria. β‐Defensin 3 has a specific role in protecting the skin. This study quantified the minimal inhibitory concentration (MIC) of human (h)BD3 against Staphylococcus pseudintermedius isolates from atopic and healthy dogs. Single colony isolates (1 × 105 colony‐forming units/mL log phase) were cultured with doubling dilutions of hBD3 in sodium phosphate buffer from 0.8 to 50 μg/mL at 37 °C for 2 h, before adding 100 μL of tryptone soy broth and incubating for a further 20 h. Bacterial growth was assessed as the mean optical density at 540 nm corrected for background. The median MIC was 12.5 μg hBD3/mL (range 3.125–25 μg/mL; n = 22). Forty‐five percent of the isolates were inhibited at ≤6.25 μg hBD3/mL, and 90% were inhibited at ≤12.5 μg hBD3/mL. Bacterial growth was not inhibited at ≤1.6 μg hBD3/mL. There were no significant differences in the inhibition by hBD3 of isolates from atopic (median MIC 12.5 μg/mL, range 6.25–25 μg/mL, n = 14) and healthy dogs (median MIC 9.4 μg/mL, range 3.125–12.5 μg/mL, n = 8); from noninfected colonized sites (median MIC 12.5 μg/mL, range 3.125–25 μg/mL, n = 16) and infected lesions (median MIC 9.4 μg/mL, range 6.25–12.5 μg/mL, n = 6); or between sample sites (nose median MIC 12.5 μg/mL, range 6.25–25 μg/mL, n = 5; perineum median MIC 12.5 μg/mL, range 3.125–25 μg/mL, n = 7; ear median MIC 6.25 μg/mL, range 6.25–12.5 μg/mL, n = 4; lesions median MIC 9.4 μg/mL, range 6.25–12.5 μg/mL, n = 6). In conclusion, hBD3 inhibited the growth of canine S. pseudintermedius isolates in vitro irrespective of origin.  相似文献   

15.
为了有效地治疗鸡大肠杆菌病,采用微量倍比稀释方法测定了17种抗菌药物对临床分离的豫北地区15株鸡大肠杆菌的体外最小抑菌浓度(MIC),并根据其MIC及MIC范围(MICRange)使用SPSS 13.0中Probit过程计算出17种抗菌药物的MIC50和MIC90.结果表明:多粘菌素B的抑菌作用最强,MIC50、MIC90分别为0.11、0.87 μg/mL;加替沙星的抑菌作用次之,MIC50、MIC90分别为2.53、3.88 μg/mL,其它3种药物恩诺沙星、左氧氟沙星、环丙沙星的抑菌作用相当,但不及加替沙星,MIC50、MIC90分别为10.11~11.79 μg/mL、15.16~21.13 μg/mL;多西环素和阿莫西林等12种抗菌药物的抑菌作用较小,MIC50、MIC90分别为18.53~388.50 μg/mL和30.59~713.42 μg/mL.  相似文献   

16.
豫北地区临床分离鸡大肠杆菌的体外抑菌作用测定   总被引:1,自引:0,他引:1  
为了有效地治疗鸡大肠杆菌病,采用微量倍比稀释方法测定了17种抗菌药物对临床分离的豫北地区15株鸡大肠杆菌的体外最小抑菌浓度(MIC),并根据其MIC及MIC范围(MICRange)使用SPSS 13.0中Probit过程计算出17种抗菌药物的MIC50和MIC90。结果表明:多粘菌素B的抑菌作用最强,MIC50、MIC90分别为0.11、0.87μg/mL;加替沙星的抑菌作用次之,MIC50、MIC90分别为2.53、3.88μg/mL,其它3种药物恩诺沙星、左氧氟沙星、环丙沙星的抑菌作用相当,但不及加替沙星,MIC50、MIC90分别为10.11-11.79μg/mL、15.16-21.13μg/mL;多西环素和阿莫西林等12种抗菌药物的抑菌作用较小,MIC50、MIC90分别为18.53-388.50μg/mL和30.59-713.42μg/mL。  相似文献   

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