bCAT通过减少白色念珠菌的粘附抑制生物被膜形成的机制研究

发布时间：2018-05-17 00:14

  本文选题：白色念珠菌 + 生物被膜形成　； 参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:明确bCAT是否具有抑制白色念珠菌生物被膜形成的作用,并探索该作用与粘附基因HWP1表达的关系。方法白色念珠菌标准菌株ATCC10231及临床菌株作为研究对象,XTT法检测其形成生物被膜的能力,通过CLSI-M27-A3方法确定b CAT抗浮游状态MIC值,XTT法及菌落计数检测bCAT抑制生物被膜形成作用,并计算代谢活性确定BIC50,bCAT减少白色念珠菌的粘附作用经倒置显微镜下观察及菌落计数法确定,并采用RT-PCR检测HWP1的表达,空白对照组和实验组使用独立样本t检验,并通过2-ΔΔCt法计算相对表达量。统计方法为单因素方差分析,组内比较采用Dunnett T3检验。结果:标准菌株及临床菌株均有强生物被膜形成能力。b CAT抗浮游状态的白色念珠菌MIC值为40-80μmol/L,抑制白色念珠菌生物被膜形成BIC50为80-160μmol/L;并且bCAT能减少白色念珠菌的粘附作用,空白对照组菌落计数为27822.22±2472.74 cfu,bCAT浓度为160、80、40、20、10μmol/L时的菌落个数分别为5355.55±1264.03 cfu、11377.78±2232.58 cfu、17488.89±1136.27 cfu、22377.78±3521.99 cfu、26044.44±1329.57 cfu。组间差异具有统计学差异(F=147.018,P=0.000),组内比较发现160、80、40μmol/L处理组与不含bCAT时差异具有统计学意义(P0.05)。RT-PCR发现160μmol/L处理组HWP1相对表达量为不含bCAT的空白对照组的12.24±11.55%,差异具有统计学意义(t=58.985,P0.05)。结论:bCAT能够有效抑制白色念珠菌形成生物被膜,其机制可与降低HWP1基因的表达从而减少白色念珠菌的粘附有关。以上研究结果为了解CGA衍生的天然抗菌多肽的先天性免疫作用增加了新的数据,并且为临床白色念珠菌耐药的问题提供了新的解决思路。
[Abstract]:Aim: to investigate whether bCAT can inhibit the formation of biofilm in Candida albicans and to explore the relationship between this effect and the expression of adhesion gene HWP1. Methods the ability of Candida albicans standard strain ATCC10231 and clinical strains to form biofilm was determined by CLSI-M27-A3 method, and the inhibitory effect of bCAT on biofilm formation was determined by CLSI-M27-A3 method. The metabolic activity was calculated to determine the effect of BIC50 BCAT on reducing the adhesion of Candida albicans by inverted microscope and colony counting method. The expression of HWP1 was detected by RT-PCR, and the independent sample t test was used in the blank control group and the experimental group. The relative expression was calculated by 2-螖 Ct method. The statistical method was single factor ANOVA and Dunnett T3 test was used for intra-group comparison. Results: both standard strain and clinical strain had strong biofilm forming ability. The MIC value of Candida albicans in anti-planktonic state was 40-80 渭 mol / L, the inhibition of Candida albicans biofilm formation BIC50 was 80-160 渭 mol / L, and bCAT could reduce the adhesion of Candida albicans. In the blank control group, the colony count was 27822.22 卤2472.74 cfujib cat and the number of the colonies was 5355.55 卤1264.03 mol/L, 11377.78 卤2232.58 cfufun 17488.89 卤1136.27 cfutio 22377.78 卤3521.99 cfun 26044.44 卤1329.57 cfu. respectively. There was significant difference in the relative expression of HWP1 between the 160 渭 mol/L treatment group and the control group without bCAT. The relative expression of HWP1 in the 160 渭 mol/L treatment group was 12.24 卤11.555.The difference was statistically significant between the 160 渭 mol/L treatment group and the control group without bCAT, and the difference was statistically significant (P 0.05). Conclusion: BCAT can effectively inhibit the formation of biofilm in Candida albicans and its mechanism may be related to the reduction of the expression of HWP1 gene and the reduction of adhesion of Candida albicans. These results provide new data for understanding the innate immune function of natural antimicrobial peptides derived from CGA and provide a new way to solve the problem of clinical resistance of Candida albicans.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R519.3

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