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目的:研究白鲜皮提取物在体外对白念珠菌生物膜及相关基因的影响。方法:M27-A2测定白鲜皮提取物对白念珠菌的最小抑菌浓度(MIC);XTT法评价白鲜皮提取物对白念珠菌生物膜形成及细胞黏附的影响;实时荧光定量RT-PCR(qRT-PCR)检测白鲜皮提取物作用后,ALS3、HWP1基因表达情况。结果:白鲜皮提取物对白念珠菌的MIC为128μg/m L;对生物膜的SMIC50和SMIC80分别为256μg/m L和512μg/m L;与空白对照组相比,不同浓度白鲜皮提取物(16~512μg/m L)对培养1、2、3、4 h的白念珠菌细胞黏附均有明显抑制作用(P<0.05);qRT-PCR结果显示药物作用后ALS3、HWP1基因表达降低(P<0.05)。结论:白鲜皮提取物对体外白念珠菌生物膜有较明显的抑制作用,其机制可能通过抑制ALS3、HWP1等相关基因的表达,从而抑制Ras/c AMP通路的功能。
OBJECTIVE: To study the effects of extract of Cortex Fragrans on C. albicans biofilms and related genes in vitro. Methods: The minimal inhibitory concentration (MIC) of Candida albicans was determined by M27-A2. The XTT method was used to evaluate the effect of P. crispus extract on the biofilm formation and cell adhesion of Candida albicans. Real-time fluorescent quantitative RT-PCR qRT-PCR) was used to detect the expression of ALS3 and HWP1 gene after the fresh-skin extract was taken. Results: The MIC for C. albicans was 128 μg / mL for C. albicans and 256 μg / mL for SMIC80 and 512 μg / mL for SM biofilms, respectively. Compared with the blank control group, (16 ~ 512μg / m L) inhibited the cell adhesion of Candida albicans cells cultured for 1, 2, 3 and 4 hours (P <0.05). The results of qRT-PCR showed that the expression of ALS3 and HWP1 mRNA decreased (P <0.05). CONCLUSION: The extract of P. crispus obviously inhibits the Candida albicans biofilm in vitro. The mechanism may inhibit the function of Ras / c AMP pathway by inhibiting the expression of related genes such as ALS3 and HWP1.