法尼醇对白念珠菌生物膜耐药性作用的研究
发布时间：2019-05-28 17:00
【摘要】：目的：本研究主要研究法尼醇对白念珠菌生物膜药物易感性、形态、麦角甾醇的作用,探讨法尼醇作为一种抗微生物化学活性分子对白念珠菌生物膜耐药性的作用。 方法：本研究分为白念珠菌标准株未处理组(标准株组、对照组)、标准株法尼醇处理组、耐药株未处理组(耐药株组)及耐药株法尼醇处理组,针对白念珠菌不同时相(6h、12h、24h、36h)生物膜进行研究。。研究采用下述方法探讨法尼醇对白念珠菌生物膜耐药相关生物学特点的作用：XTT分析检测不同浓度法尼醇对白念珠菌生物膜的抑制作用；XTT分析用于不同生物膜时相法尼醇作用于白念珠菌生物膜的药物易感性实验；扫描电镜观察比较法尼醇处理后白念珠菌菌体形态的改变,激光共聚焦比较各组生物膜厚度；液相色相质谱联用仪对各组细胞质膜成分进行检测并比较。 结果：实验一：在生物膜形成6、12h,标准株和耐药株高浓度法尼醇处理组抑菌率比低浓度法尼醇处理组高；而标准株法尼醇处理组和耐药株法尼醇处理组的24h生物膜在法尼醇处理浓度为0-150μM时,抑菌率没有明显升高。标准株法尼醇处理组在法尼醇处理浓度为50μM时,四个时相生物膜的抑菌率无明显改变；在法尼醇处理浓度为100、150、200、250、300μM时,同一法尼醇浓度作用于标准株法尼醇处理组四个时相生物膜,抑制作用随时间延长而减弱。在同一生物膜时相,同一浓度法尼醇对耐药株生物膜的抑制作用弱于标准株。 实验二：XTT法检测白念珠菌(标准株组、标准株法尼醇处理组、耐药株组及耐药株法尼醇处理组)生物膜MIC,发现标准株法尼醇处理组生物膜SMICgo在6h、12h、24h、36h均低于标准株组,且差异在36h变小；耐药株法尼醇处理组生物膜SMIC80在6h、12h、24h低于耐药株组,在36h,四组生物膜SMIC80均无明显差异。耐药株生物膜SMIC80在6h、12h、24h及36h均高于标准株生物膜。 实验三：扫描电镜观察四组白念珠菌生物膜的附着状态及菌体形态,发现法尼醇处理使白念珠菌菌丝形成较晚且菌丝变短,这个变化在6h、12h和24h时较为明显,在36h则变化不明显。法尼醇使白念珠菌菌体表面皱缩和形成凹陷,在生物膜早期及中期尤为显著,随着生物膜的成熟,凹陷逐渐变少,至生物膜成熟期凹陷已经较为少见。同一时段下耐药株法尼醇处理组菌体表面的凹陷少于标准株法尼醇处理组。同一时段下耐药株组细胞外基质多于标准株法尼醇处理组。激光共聚焦观察发现,在生物膜形成6h、12h、24h,标准株组生物膜厚度高于标准株法尼醇处理组,耐药株组生物膜厚度高于耐药株法尼醇处理组。在36h,法尼醇处理后生物膜厚度未显著降低。在6h及12h,耐药株生物膜厚度高于标准株。 实验四：HPLC检测分析麦角甾醇结果显示,在生物膜形成6h、12h、24h,同一时段法尼醇处理组生物膜麦角甾醇含量低于未处理组,标准株法尼醇处理组生物膜麦角甾醇含量均低于标准株组,耐药株法尼醇处理组生物膜麦角甾醇含量均低于耐药株组,且耐药株组麦角甾醇含量高于标准株组。 随着生物膜的逐渐成熟,标准株组生物膜麦角甾醇含量逐渐升高,标准株法尼醇处理组24h生物膜麦角甾醇含量较6h生物膜升高,标准株法尼醇处理组36h生物膜麦角甾醇含量较24h生物膜显著降低；耐药株组24h及36h生物膜麦角甾醇含量较6h生物膜显著降低。 结论： 实验一：法尼醇对白念珠菌生物膜的抑制作用与法尼醇浓度及白念珠菌的生物膜时相相关。在生物膜早期及中期,高浓度法尼醇对生物膜的抑制效果比低浓度法尼醇强；在生物膜成熟期,法尼醇对白念珠菌生物膜无明显抑制作用。法尼醇对白念珠菌各时期生物膜的抑制作用随生物膜成熟而减弱,对早期生物膜抑制作用最强。 实验二：法尼醇可以抑制白念珠菌生物膜耐药性,且法尼醇对早期及中期生物膜耐药性的抑制作用明显,对晚期成熟生物膜耐药性的抑制作用不如早期及中期明显。 实验三：研究结果显示法尼醇抑制白念珠菌胞外基质的形成,造成菌体表面损伤,抑制菌丝的形成和伸长,影响生物膜的形成,降低生物膜厚度。法尼醇的这些作用对早期和中期生物膜较明显,对成熟期生物膜无明显作用。 实验四：麦角甾醇含量与耐药性呈正相关。法尼醇能降低早期及中期白念珠菌生物膜中的麦角甾醇含量,推测法尼醇抑制白念珠菌生物膜耐药性的机制是法尼醇降低生物膜中的麦角甾醇含量,使细胞质膜稳定性下降,对唑类药物通透性改变,从而导致白念珠菌对唑类药物的耐药性下降。
[Abstract]:Objective: To study the effect of farnesol on the drug susceptibility, morphology and ergoline, and to explore the effect of farnesol as an anti-microbial chemical active molecule on the drug resistance of Candida albicans. Methods: The study was divided into two groups: non-treated group (standard plant group, control group), standard strain FENOL treatment group, resistant strain non-treatment group (drug resistant strain group) and drug-resistant strain method, and the non-simultaneous phase of Candida albicans (6h, 12h,24). h,36 h) Biofilm ............................................................................................................ Application of XTT in the study of drug susceptibility to the membrane of Candida albicans in different biological membrane phases; the changes of the morphology of Candida albicans after the treatment with the method of scanning electron microscopy; and the thickness of the biofilms in each group were compared by laser confocal. The plasma membrane components of each group were detected and compared by a liquid phase-phase-phase-mass-mass spectrometer. The results were as follows:1. The inhibition rate of the high-concentration farnesol treatment group was higher than that of the low-concentration farnesol treatment group in the biofilm formation of 6,12 h, the standard strain and the drug-resistant strain. The inhibition rate is not clear at the time of the. m The results showed that the inhibition rate of the four-phase biofilms was not significantly changed when the treatment concentration of the normal plant was 50. m u.M. The concentration of the same method was 100,150,200,250,300. m u.M, and the concentration of the same method in the treatment group of the normal strain was 100,150,200,250 and 300. m Biofilm, inhibition over time In the same biological membrane phase, the inhibition of the same concentration of farnesol on the biofilms of drug-resistant strains was weaker than that of the same biofilms. Standard strain was used to test the MIC of the biofilms of Candida albicans (standard plant group, standard strain Fanyol treatment group, drug-resistant strain group and drug-resistant strain Fanyol treatment group) by means of XTT method, and it was found that the biofilms SMICgo of the standard strain method were lower than that of the standard plant group at 6 h,12 h,24 h and 36 h, and the difference was The treatment group biofilm SMIC80 was lower than that of the resistant strain group at 6 h,12 h and 24 h. There was no significant difference. The biological membrane of the resistant strain SMIC80 was higher than the standard in 6 h,12 h,24 h and 36 h. The attachment status and the cell morphology of the four groups of Candida albicans biofilm were observed by scanning electron microscope. The results showed that the hyphae of Candida albicans were formed later and the hyphae became shorter at 6 h,12 h and 24 h, and at 36 h,12 h and 24 h, In that early and mid-term of the biofilms, with the mature of the biofilms, the depression became less, and the maturation of the biofilms was concave. The depression of the cell surface of the drug-resistant strain in the same period is less than that of the standard. The extracellular matrix of drug-resistant strains in the same period was more than that of the standard. It was found that the membrane thickness of the standard plant group was higher than that of the standard method, the thickness of the biofilm was higher than that of the standard strain, the thickness of the biofilm was higher than that of the standard strain, the thickness of the biofilm was higher than that of the standard strain. Plant farnesol treatment group. After 36 h, the biofilms were treated with farnesol. The thickness was not significantly reduced. In 6 h and 12 h, the biofilms of the resistant strains The results showed that the content of ergoline in the treatment group was lower than that of the untreated group in the same period of time, and the results showed that the content of the ergoline in the treatment group was lower than that of the untreated group. The content of ergoline was lower than that of the standard plant, and the content of ergoline in the treatment group of the drug-resistant strain was lower than that of the resistant strains. The alcohol content was higher than that of the standard plant group. With the gradual maturation of the biofilms, the content of ergoline in the standard plant group was gradually increased, and the content of the ergoline in the standard strain method of the treatment group was higher than that of the 6-h biofilms. The content of ergoline in the treatment group of the standard strain in the treatment group for 36h was higher than that of the standard plant group. The 24-hour and 36-hour biofilms of the drug-resistant strains were significantly lower than that of the 24-hour biofilms. The amount is 6 Conclusion: The inhibitory effect of farnesol on the biological membrane of Candida albicans and the inhibition of farnesol In the early and mid-term of the biofilms, the inhibition effect of the high-concentration farnesol on the biofilms is stronger than that of the low-concentration farnesol; in the mature period of the biofilms, the farnesol The inhibitory effect of farnesol on the biofilms of Candida albicans in various stages of Candida albicans was observed with the maturation of the biofilms. The results showed that the inhibition of the drug resistance of Candida albicans biofilm was inhibited, and the inhibition of the drug resistance of farnesol on the early and medium-term biofilm was evident. The results showed that the formation of extracellular matrix of Candida albicans was inhibited, the surface of the cells was damaged, and the formation of hyphae was inhibited. elongation, the formation of biofilm, and the reduction of biofilm thickness. These effects of farnesol on early and mid-term The biofilm is more obvious and has no obvious effect on the mature membrane of the mature period. The experimental results showed that the content of ergoline was positively correlated with the drug resistance. The results showed that the content of ergoline in the early and medium-term Candida albicans biofilm was reduced, and the mechanism of the inhibition of the drug resistance of the farnesol on the biofilms of Candida albicans was the decrease of the content of ergoline in the biofilm. the stability of the cytoplasmic membrane is reduced, the permeability of the plasma membrane is modified,
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R780.2
本文编号：2487219
[Abstract]:Objective: To study the effect of farnesol on the drug susceptibility, morphology and ergoline, and to explore the effect of farnesol as an anti-microbial chemical active molecule on the drug resistance of Candida albicans. Methods: The study was divided into two groups: non-treated group (standard plant group, control group), standard strain FENOL treatment group, resistant strain non-treatment group (drug resistant strain group) and drug-resistant strain method, and the non-simultaneous phase of Candida albicans (6h, 12h,24). h,36 h) Biofilm ............................................................................................................ Application of XTT in the study of drug susceptibility to the membrane of Candida albicans in different biological membrane phases; the changes of the morphology of Candida albicans after the treatment with the method of scanning electron microscopy; and the thickness of the biofilms in each group were compared by laser confocal. The plasma membrane components of each group were detected and compared by a liquid phase-phase-phase-mass-mass spectrometer. The results were as follows:1. The inhibition rate of the high-concentration farnesol treatment group was higher than that of the low-concentration farnesol treatment group in the biofilm formation of 6,12 h, the standard strain and the drug-resistant strain. The inhibition rate is not clear at the time of the. m The results showed that the inhibition rate of the four-phase biofilms was not significantly changed when the treatment concentration of the normal plant was 50. m u.M. The concentration of the same method was 100,150,200,250,300. m u.M, and the concentration of the same method in the treatment group of the normal strain was 100,150,200,250 and 300. m Biofilm, inhibition over time In the same biological membrane phase, the inhibition of the same concentration of farnesol on the biofilms of drug-resistant strains was weaker than that of the same biofilms. Standard strain was used to test the MIC of the biofilms of Candida albicans (standard plant group, standard strain Fanyol treatment group, drug-resistant strain group and drug-resistant strain Fanyol treatment group) by means of XTT method, and it was found that the biofilms SMICgo of the standard strain method were lower than that of the standard plant group at 6 h,12 h,24 h and 36 h, and the difference was The treatment group biofilm SMIC80 was lower than that of the resistant strain group at 6 h,12 h and 24 h. There was no significant difference. The biological membrane of the resistant strain SMIC80 was higher than the standard in 6 h,12 h,24 h and 36 h. The attachment status and the cell morphology of the four groups of Candida albicans biofilm were observed by scanning electron microscope. The results showed that the hyphae of Candida albicans were formed later and the hyphae became shorter at 6 h,12 h and 24 h, and at 36 h,12 h and 24 h, In that early and mid-term of the biofilms, with the mature of the biofilms, the depression became less, and the maturation of the biofilms was concave. The depression of the cell surface of the drug-resistant strain in the same period is less than that of the standard. The extracellular matrix of drug-resistant strains in the same period was more than that of the standard. It was found that the membrane thickness of the standard plant group was higher than that of the standard method, the thickness of the biofilm was higher than that of the standard strain, the thickness of the biofilm was higher than that of the standard strain, the thickness of the biofilm was higher than that of the standard strain. Plant farnesol treatment group. After 36 h, the biofilms were treated with farnesol. The thickness was not significantly reduced. In 6 h and 12 h, the biofilms of the resistant strains The results showed that the content of ergoline in the treatment group was lower than that of the untreated group in the same period of time, and the results showed that the content of the ergoline in the treatment group was lower than that of the untreated group. The content of ergoline was lower than that of the standard plant, and the content of ergoline in the treatment group of the drug-resistant strain was lower than that of the resistant strains. The alcohol content was higher than that of the standard plant group. With the gradual maturation of the biofilms, the content of ergoline in the standard plant group was gradually increased, and the content of the ergoline in the standard strain method of the treatment group was higher than that of the 6-h biofilms. The content of ergoline in the treatment group of the standard strain in the treatment group for 36h was higher than that of the standard plant group. The 24-hour and 36-hour biofilms of the drug-resistant strains were significantly lower than that of the 24-hour biofilms. The amount is 6 Conclusion: The inhibitory effect of farnesol on the biological membrane of Candida albicans and the inhibition of farnesol In the early and mid-term of the biofilms, the inhibition effect of the high-concentration farnesol on the biofilms is stronger than that of the low-concentration farnesol; in the mature period of the biofilms, the farnesol The inhibitory effect of farnesol on the biofilms of Candida albicans in various stages of Candida albicans was observed with the maturation of the biofilms. The results showed that the inhibition of the drug resistance of Candida albicans biofilm was inhibited, and the inhibition of the drug resistance of farnesol on the early and medium-term biofilm was evident. The results showed that the formation of extracellular matrix of Candida albicans was inhibited, the surface of the cells was damaged, and the formation of hyphae was inhibited. elongation, the formation of biofilm, and the reduction of biofilm thickness. These effects of farnesol on early and mid-term The biofilm is more obvious and has no obvious effect on the mature membrane of the mature period. The experimental results showed that the content of ergoline was positively correlated with the drug resistance. The results showed that the content of ergoline in the early and medium-term Candida albicans biofilm was reduced, and the mechanism of the inhibition of the drug resistance of the farnesol on the biofilms of Candida albicans was the decrease of the content of ergoline in the biofilm. the stability of the cytoplasmic membrane is reduced, the permeability of the plasma membrane is modified,
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R780.2
【参考文献】
相关期刊论文 前1条
1 王应斌,王宏,郭辉玉,赵永忠,罗深秋;白假丝酵母ERG11基因突变分析[J];第一军医大学学报;2005年11期
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