大蒜素对白念珠菌形态转换的影响研究

摘要/Abstract

摘要： 目的探讨大蒜素对白念珠菌形态转换的影响及其作用机制。方法倒置显微镜观察白念珠菌菌丝形成的体外动力学过程；采用CLSI-M27-A3微量液基稀释法检测大蒜素对白念珠菌的最小抑菌浓度（minimum inhibitory concentration，MIC）；倒置显微镜观察不同浓度大蒜素对白念珠菌在Spider液体培养基中菌丝形成的影响；qRT-PCR法检测在不同浓度大蒜素作用下白念珠菌菌丝相关基因HWP1、ALS1、EFG1、PDE2表达水平的变化。结果白念珠菌在Spider液体培养基中6 h时出现较长菌丝，24 h后镜下可见大量念珠菌菌丝包裹酵母细胞，紧密交错；大蒜素对白念珠菌的MIC值为25 μg/mL；倒置显微镜观察（25~100）μg/mL浓度的大蒜素能明显抑制Spider液体培养基中白念珠菌菌丝的生长；qRT-PCR结果显示，在（25~100）μg/mL浓度的大蒜素作用下，白念珠菌菌丝相关基因表达下调。结论大蒜素能有效抑制白念珠菌的形态转换，其作用机制可能与调节菌丝形成相关基因的表达水平有关。

关键词: 白念珠菌, 形态转换, 菌丝相, 大蒜素

Abstract: Objective To investigate the effect and mechanism of allicin on morphological transition of Candida albicans. Methods The inverted process of dynamic formation of Candida albicans hyphae was observed under microscope. CLSI-M27-A3 microdilution method was used to detect the minimum inhibitory concentration(MIC)of allicin against Candida albicans. The effect of allicin on the hyphal formation of Candida albicans in Spider liquid medium was observed under inverted microscope. qRT-PCR method was used to measure the expression of hypal related genes HWP1,ALS1,EFG1 and PDE2 of Candida albicans under the effect of allicin. Results Long hyphae began to appear in the Spider liquid medium at 6h. After 24 hours, a large number of hyphal wrapped yeast cells which interlaced tightly could be seen under inverted microscope. The MIC of allicin against to Candida albicans was 25 μg/mL. The growth of Candida albicans hyphae could be significantly inhibited by allicin in concentrations of 25-100 μg/mL in Spider liquid media. qRT-PCR results showed that the expression of hyphae related genes was down-regulated under the effect of allicin in concentrations of 25-100 μg/mL. Conclusion Allicin could significantly inhibit the morphological transition of Candida albicans, and the mechanism may correlate to the regulation of the expression level of the genes which related to hyphal formation.

Key words: Candida albicans, morphological transition, hyphal form, allicin

中图分类号:

R379.4

熊延靖, 厉荣玉, 吴艳红. 大蒜素对白念珠菌形态转换的影响研究[J]. 中国真菌学杂志, 2021, 16(1): 14-18.

XIONG Yanjing, LI Rongyu, WU Yanhong. The effect of allicin on the morphological transition of Candida albicans[J]. Chinese Journal of Mycology, 2021, 16(1): 14-18.

参考文献

[1] Nobile CJ, Johnson AD. Candida albicans biofilms and human disease[J]. Annu Rev Microbiol, 2015, 69:71-92.
[2] Zeng B, Li J, Wang Y, et al. In vitro and in vivo effects of suloctidil on growth and biofilm formation of the opportunistic fungus Candida albicans[J]. Oncotarget, 2017, 8(41):69972-69982.
[3] Li D, Bernhardt J, Calderone R. Temporal expression of the Candida albicans genes CHK1 and CSSK1, adherence, and morphogenesis in a model of reconstituted human esophageal epithelial candidiasis[J]. Infect Immun, 2002, 70:1558-1565.
[4] Whiteway M, Oberholzer U. Candida morphogenesis and host-pathogen interactions[J]. Curr Opin Microbiol, 2004, 7(4):350-357.
[5] Naseem S, Araya E, Konopka JB, et al. Hyphal growth in Candida albicans does not require induction of hyphal-specific gene expression[J]. Mol Bio Cell, 2015, 26(6):1174-1187.
[6] 马弋, 朱必婷, 王田. 大蒜素对副溶血弧菌抑菌作用的研究[J]. 公共卫生与预防医学, 2017, 28(1):130-132.
[7] 刘娟, 谢琳, 任琪琪, 等. 大蒜汁的抑菌作用研究[J]. 湖南师范大学学报(医学版), 2019,16(4):1-3.
[8] 熊延靖, 董群. 大蒜素对体外白色念珠菌生物活性影响的研究[J]. 中国临床药理学与治疗学, 2015, 20(11):1233-1238.
[9] CLSI. Reference method for broth dilution antifungal susceptibility testing of yeast[S]. 4^th Ed. CLSI Standard M27. Wanyne, PA:Clinical Laboratory Standards Institute,2017.
[10] 李颖. 两类天然产物的抗真菌活性及作用机制研究[D]. 济南:山东大学, 2016:101-102.
[11] 陆一纶, 吴岚, 唐若愚, 等. 白念珠菌毒力因子及致病机制研究进展[J]. 中国真菌学杂志, 2018, 13(2):101-104.
[12] 朱丹婷, 李景, 俞立英. 白念珠菌致病性研究进展[J]. 中国真菌学杂志, 2015, 10(1):54-58.
[13] Moyes DL, Wilson D, Richardson JP, et al. Candidalysin is a fungal peptide toxin critical for mucosal infection[J]. Nature, 2016, 532(7597):64-68.
[14] Sudbery PE. Growth of Candida albicans hyphae[J]. Nat Rev Microbiol, 2011, 9(10):737-748.
[15] 贺慧娟, 谢水祥. 中草药抗白色念珠菌的作用机制[J]. 赣南医学院学报, 2017, 37(6):979-981.
[16] 彭敏. 大蒜素药理学作用机制及研究进展[J]. 现代中西医结合杂志,2018, 27(14):1593-1596.
[17] 逢金花, 唐建国, 童译庆, 等. 白念珠菌酵母相-菌丝相形态转换机制的研究进展[J]. 现代生物医学进展, 2017, 17(35):6950-6954.
[18] De Oliveira JR, Figueira LW, Sper FL, et al. Thymus vulgaris L. and thymol assist murine macrophages (RAW 264.7) in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans[J]. Immunol Res, 2017, 65(4):932-943.
[19] Hogan DA, Sundstrom P. The Ras/cAMP/PKA signaling pathway and virulence in Candida albicans[J]. Future Microbiol, 2009, 4(10):1263-1270.