黄芩苷联合氟康唑通过降低cAMP抑制白念珠菌酵母-菌丝形态转化

中国真菌学杂志　 2016, Vol. 11 　Issue (1): 8-12.

黄芩苷联合氟康唑通过降低cAMP抑制白念珠菌酵母-菌丝形态转化

冯鑫, 施高翔, 云云, 吴大强, 邵菁, 汪天明, 汪长中

安徽中医药大学中西医结合临床学院安徽省中医药科学院中西医结合研究所, 合肥 230038

收稿日期:2016-01-15 出版日期:2016-02-28 发布日期:2016-02-28
通讯作者: 汪长中,E-mail:ahwcz63@sina.com E-mail:ahwcz63@sina.com
作者简介:冯鑫,男(回族),硕士,讲师.E-mail:fx74@163.com
基金资助:
安徽省自然科学基金项目(1408085MH165,1508085MH163)

Effect of baicalin in combination with fluconazole against Candida albicans morphological transition from yeast to hyphae via inhibiting cAMP

FENG Xin, SHI Gao-xiang, YUN Yun, WU Da-qiang, SHAO Jing, WANG Tian-ming, WANG Chang-zhong

School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Institute of Integrated Traditional and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230038

Received:2016-01-15 Online:2016-02-28 Published:2016-02-28

摘要/Abstract

摘要：

目的探讨中药有效成分黄芩苷(Baicalin,BA)联合氟康唑(Fluconazole,FLC)对白念珠菌(Candida albicans)形态转化的影响及机制。方法在菌丝诱导培养基(液体和固体培养基)上观察两药联用对白念珠菌酵母-菌丝形态转化的影响;ELISA法检测两药联用对胞内cAMP含量的影响;回补实验验证两药联用对胞内cAMP的影响;qRT-PCR检测两药联用对白念珠菌cAMP相关基因RAS1,CDC35和PDE2表达的影响。结果黄芩苷联合氟康唑能显著抑制白念珠菌酵母-菌丝形态转化,降低胞内cAMP水平;外源性cAMP能逆转两药联用所造成的菌丝抑制;两药联用使RAS1和CDC35表达分别下调35%和27%,使PDE2上调1.21倍。结论黄芩苷协同氟康唑显著抑制白念珠菌形态转化,其作用机制可能与下调胞内cAMP有关。

关键词: 白念珠菌, 黄芩苷, 氟康唑, 形态转化, cAMP

Abstract:

Objective This study aimed to investigate the antifungal activity of baicalin in combination with fluconazole against Candida albicans morphological transition from yeast to hyphae and to explore the mechanism.Methods The hypha-inducing liquid and solid media were uesd to observe the effect of baicalin alone or in combination with fluconazole on yeast-hapal transformation in C.albicans;The intracellular cAMP level and cAMP rescue was measured by ELISA Kit;Quantitative real time PCR(qRT-PCR) was performed to measure the expression of cAMP-related genes.Results Baicalin in combination with fluconazole could inhibit hyphae formation and intracellular cAMP level obviously;exogenous cAMP could convert hypha-inhibiting effect;And expression of RAS1 and CDC35 were downregulated by 35% and 27% respectively,while the expression of PDE2 was upregulated by 121%.Conclusion Mechanism of baicalin in combination with fluconazole inhibiting C.albicans morphological transition was probabaly via down-regulating the intracellular cAMP.

Key words: C.albicans, baicalin, fluconazole, morphological transition, cAMP

中图分类号:

R379.4

冯鑫, 施高翔, 云云, 吴大强, 邵菁, 汪天明, 汪长中. 黄芩苷联合氟康唑通过降低cAMP抑制白念珠菌酵母-菌丝形态转化[J]. 中国真菌学杂志, 2016, 11(1): 8-12.

FENG Xin, SHI Gao-xiang, YUN Yun, WU Da-qiang, SHAO Jing, WANG Tian-ming, WANG Chang-zhong. Effect of baicalin in combination with fluconazole against Candida albicans morphological transition from yeast to hyphae via inhibiting cAMP[J]. Chinese Journal of Mycology, 2016, 11(1): 8-12.

参考文献

[1] Martin GS,Maimino DM,Eaton S,et al.The epidemiology of sepsis in the United States from 1979 through 2000[J].N Eng J Med,2003,348(16):1546-1554.
[2] Carlisle PL,Banerjee M,Lazzell A,et al.Expression levels of a filament-specific transcriptional regulator are sufficient to determine Candida albicans morphology and virulence[J].Proc Natl Acad Sci U S A,2009,106(2):599-604.
[3] Chou TC,Chang LP,Li CY,et al.Theamtiinflammatory and analgesic effects of baicalin in carrageenan-evoked thermal hyperalgesia[J].Ansth Analg,2003,97(6):1724-1729.
[4] 熊英,傅颖媛,况南珍,等.黄芩苷抗白念珠菌作用机制研究[J],中国药理学通报,2004,20(12):1404-1407.
[5] Wang TM,Shi GX,Shao J,et al.In vitro antifungal activity of baicalin against Candida albicans biofilms via apoptotic induction[J].Micro Patho,2015,87:21-29.
[6] 颜贵明,施高翔,邵菁,等.黄芩苷联合氟康唑对白念珠菌生物膜的抑制作用研究[J].中国真菌学杂志,2015,10(3):139-145.
[7] 陈新辉,赵明玄,安金刚,等.单克隆天然抗角蛋白抗体IgM3B4体外抑制白念珠菌芽管形成及黏附作用[J].第四军医大学学报,2008,29(1):47-50.
[8] Maidan MM,De Rop L,Semeels J,et al.The G protein-coupled receptor Gprl and the Ga protein Gpa2 act through the cAMP-protein kinase A pathway to induce morphogenesis in Candida albicans[J].Mol Biol Cell,2005,16(4):1971-1986.
[9] Jung WH,Stateva LI.The cAMP phosphodiesterase encoded by CaPDE2 is required for hyphal development in Candida albicans[J].Microbiology,2003,149(Pt 10):2961-2976.
[10] Xie Z,Thompson A,Kashleva H,et al.A quantitative real-time RT-PCR assay for mature C.albicans biofilms[J].BMC Microbiol,2011,11:93.
[11] Hogan DA,Sundstrom P.The Ras/cAMP/PKA signaling pathway and virulence in Candida albicans[J].Future Microbiol,2009,4(10):1263-1270.

[1]	黄进宝, 兰长青, 翁恒, 林清华. 肋骨隐球菌感染2例报道并文献复习[J]. 中国真菌学杂志, 2020, 15(5): 278-282.
[2]	刘丹, 邢丽枝, 杨心茹, 罗伟, 何荣霞. 唑类药物治疗外阴阴道假丝酵母菌病研究进展[J]. 中国真菌学杂志, 2020, 15(5): 318-320.
[3]	马玉帛, 张逸, 黄晨, 王晓雯, 李若瑜. 自然杀伤细胞在真菌感染免疫的研究进展[J]. 中国真菌学杂志, 2020, 15(3): 175-178.
[4]	邱熙然, 陈思敏, 侯炜彤, 张玉, 郭诗雨, 姜远英, 安毛毛. 白念珠菌侵袭宿主毒力因子研究进展[J]. 中国真菌学杂志, 2020, 15(3): 183-188.
[5]	张静云, 彭靖雯, 王琼, 高盈, 陈万鑫, 沈永年, Li Dong-mei, 佘晓东, 刘维达. 表皮生长因子受体在阴道念珠菌病发病过程中的作用初探[J]. 中国真菌学杂志, 2020, 15(2): 65-71.
[6]	刘光荣, 翟春涛, 金敏蓉, 李成亮. 天然抗菌剂对5种致病菌的抑制作用研究[J]. 中国真菌学杂志, 2020, 15(2): 88-92.
[7]	谭静文, 徐红, 宋金凤, 高志琴, 杨虹, 杨连娟, 温海. 聚六亚甲基双胍体外抗真菌作用研究[J]. 中国真菌学杂志, 2020, 15(2): 93-96.
[8]	张静云, 佘晓东, 刘维达. 白念珠菌糖代谢与毒力[J]. 中国真菌学杂志, 2020, 15(1): 61-64.
[9]	刘文丽, 吴迪, 陈爱明, 苏万通, 鲍邦龙. 热带念珠菌致下肢皮肤感染1例并文献复习[J]. 中国真菌学杂志, 2019, 14(6): 369-370.
[10]	赵晓琴, 冯文莉. 转录因子在白念珠菌生物膜形成过程中的调控机制[J]. 中国真菌学杂志, 2019, 14(6): 381-384.
[11]	王钰婷, 王玉柱, 刘锦燕, 孟玲宁, 李卫华, 项明洁. 酵母相和菌丝相白念珠菌感染阴道上皮细胞免疫反应的比较[J]. 中国真菌学杂志, 2019, 14(4): 193-199.
[12]	蔡晴, 杨连娟, 高志琴, 任弘瑾, 杨虹, 谭静文. 罗米地辛对氟康唑体外抗白念珠菌影响的研究[J]. 中国真菌学杂志, 2019, 14(4): 200-203,216.
[13]	葛力瑜, 梁官钊, 刘维达. 抗黏膜白念珠菌感染的固有免疫研究进展[J]. 中国真菌学杂志, 2019, 14(4): 249-252.
[14]	姬名硕, 王晓东, 帕丽达·阿布利孜. NLRP3炎症小体在真菌感染中的作用机制研究进展[J]. 中国真菌学杂志, 2019, 14(4): 253-256.
[15]	施高翔, 姜晶晶, 汪云霞, 赵婷, 邵菁, 汪天明, 汪长中. 苦参-蛇床子药对提取物对白念珠菌VVC临床株的抑制作用研究[J]. 中国真菌学杂志, 2019, 14(3): 147-153.