發(fā)布時間：2018-05-17 00:14

  本文選題：白色念珠菌 + 生物被膜形成　； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:明確bCAT是否具有抑制白色念珠菌生物被膜形成的作用,并探索該作用與粘附基因HWP1表達(dá)的關(guān)系。方法白色念珠菌標(biāo)準(zhǔn)菌株ATCC10231及臨床菌株作為研究對象,XTT法檢測其形成生物被膜的能力,通過CLSI-M27-A3方法確定b CAT抗浮游狀態(tài)MIC值,XTT法及菌落計(jì)數(shù)檢測bCAT抑制生物被膜形成作用,并計(jì)算代謝活性確定BIC50,bCAT減少白色念珠菌的粘附作用經(jīng)倒置顯微鏡下觀察及菌落計(jì)數(shù)法確定,并采用RT-PCR檢測HWP1的表達(dá),空白對照組和實(shí)驗(yàn)組使用獨(dú)立樣本t檢驗(yàn),并通過2-ΔΔCt法計(jì)算相對表達(dá)量。統(tǒng)計(jì)方法為單因素方差分析,組內(nèi)比較采用Dunnett T3檢驗(yàn)。結(jié)果:標(biāo)準(zhǔn)菌株及臨床菌株均有強(qiáng)生物被膜形成能力。b CAT抗浮游狀態(tài)的白色念珠菌MIC值為40-80μmol/L,抑制白色念珠菌生物被膜形成BIC50為80-160μmol/L;并且bCAT能減少白色念珠菌的粘附作用,空白對照組菌落計(jì)數(shù)為27822.22±2472.74 cfu,bCAT濃度為160、80、40、20、10μmol/L時的菌落個數(shù)分別為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。組間差異具有統(tǒng)計(jì)學(xué)差異(F=147.018,P=0.000),組內(nèi)比較發(fā)現(xiàn)160、80、40μmol/L處理組與不含bCAT時差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。RT-PCR發(fā)現(xiàn)160μmol/L處理組HWP1相對表達(dá)量為不含bCAT的空白對照組的12.24±11.55%,差異具有統(tǒng)計(jì)學(xué)意義(t=58.985,P0.05)。結(jié)論:bCAT能夠有效抑制白色念珠菌形成生物被膜,其機(jī)制可與降低HWP1基因的表達(dá)從而減少白色念珠菌的粘附有關(guān)。以上研究結(jié)果為了解CGA衍生的天然抗菌多肽的先天性免疫作用增加了新的數(shù)據(jù),并且為臨床白色念珠菌耐藥的問題提供了新的解決思路。
[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 鹵2472.74 cfujib cat and the number of the colonies was 5355.55 鹵1264.03 mol/L, 11377.78 鹵2232.58 cfufun 17488.89 鹵1136.27 cfutio 22377.78 鹵3521.99 cfun 26044.44 鹵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 鹵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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R519.3

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本文編號：1899034