白头翁汤正丁醇提取物对酸性条件下白念珠菌pH突变株黏附的影响  

Effects of butyl alcohol extract of Baitouweng Decoction on Candida albicans adhesion based on pH-mutant strains under acidic conditions

作　　者：汪云霞[1,2,3] 马克龙 王艳[1,2,3] 吴大强 邵菁以[3] 汪天明 汪长中[1,2,3] WANG Yun-xia;MA Ke-long;WANG Yan;WU Da-qiang;SHAO Jing;WANG Tian-ming;WANG Chang-zhong(Institute of Integrated Traditional Chinese and Western Medicine,Anhui University of Traditional Chinese Medicine,Hefei 230012,China;Institute of Integrated Traditional Chinese and Western Medicine,Anhui Academy of Traditional Chinese Medicine,Hefei 230012,China;Key Laboratory of Chinese Medicinal Formula of Anhui Province,Hefei 230012,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽省中医药科学院中西医结合研究所,安徽合肥230012 [3]中药复方安徽省重点实验室,安徽合肥230012

出　　处：《中草药》2019年第24期6009-6016,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81573725);国家自然科学基金项目(81774034);安徽中医药大学重点项目(2017fzd012);安徽省教育厅重点项目(KJ2017A301);安徽省教育厅重点项目(KJ2018A0276)

摘　　要：目的 基于pH信号通路探讨白头翁汤正丁醇提取物(butyl alcohol extract of Baitouweng Decoction,BAEB)对白念珠菌黏附的影响。方法 Spotassay检测酸性条件下pH突变株的活性;XTT法检测酸性条件下pH突变株黏附时的代谢活力;荧光显微镜观察酸性条件下pH突变株细胞黏附活力;正辛烷容纳法测定酸性条件下pH突变株疏水性;实时荧光定量PCR(qRT-PCR)检测酸性条件下pH突变株黏附相关基因的表达。结果 512μg/m L BAEB对共培养24、48 h的pH突变株的活性影响不大;phr2/phr2在酸性条件下代谢活性低,512μg/m L BAEB可明显抑制白念珠菌野生株(WT)、PHR2回补菌株、rim101/rim101、RIM101回补菌株的代谢活性,对phr2/phr2代谢活性影响不大;512μg/m L BAEB可明显抑制WT、PHR2回补菌株、rim101/rim101、RIM101回补菌株的细胞黏附活力,phr2/phr2加药前后细胞黏附活性无明显变化;512μg/mL BAEB对WT、phr2/phr2、PHR2回补、rim101/rim101及RIM101回补菌株的细胞表面疏水性影响不明显;q RT-PCR法检测512μg/m L BAEB对pH突变株在酸性条件下黏附相关基因作用不明显,但1 024μg/m L BAEB则对大多数黏附相关基因有明显抑制作用。结论 BAEB在酸性条件下能一定程度抑制白念珠菌的黏附。Objective To investigate the effect and mechanism of butyl alcohol extract of Baitouweng Decoction(BAEB)on adhesion of Candida albicans based on pH signaling pathway.Methods Spot assay method was used to detect the sensitivity of pH mutants to BAEB under acidic conditions.XTT assay was used to detect the effect of BAEB on metabolic activity of pH mutants.The effect of BAEB on the adhesion activity of pH mutants was observed by fluorescence microscopy.The effect of BAEB on hydrophobicity of pH mutant was determined by/i-octane inclusion method.The effect of BAEB on the expression of adhesion genes related to pH mutants was detected by qRT-PCR.Results Under acidic conditions,spot assay observation showed that pH mutants were less sensitive to BAEB,512 ng/mL BAEB interfered with pH mutants for 24 h and 48 h,there was no significantly decrease in bacterial colony.XTT assay showed that the metabolic activity of WT,PHR2 complementation,rimlOl/rimlOl and RIM 101 complementation was significantly inhibited in 512μg/mL BAEB,and there was no significantly difference in the inhibition of phr2/phr2 metabolic activity.Fluorescence microscopy showed that the cell adhesion activity of WT,PHR2 complementation,rimlOl/rimlOl,RIM 101 complementation was significantly inhibited in 512μg/rnL BAEB,the cell adhesion activity of phr2/phr2 had no obvious effect in 512 μg/mL BAEB.The n-octane inclusion method showed that the effect of 512 ng/mL BAEB on the cell surface hydrophobicity of WT,phr2/phr2,PHR2 complementation,rimlOl/rimlOl,RIM 101 complementation was not significant.The qRT-PCR assay showed that the adhesion genes of pH mutants was inhibited in 1024 μg/mL BAEB.Conclusion Under acidic conditions,the Candida albicans pH mutants was inhibited by BAEB to a certain extent.

关 键 词：白头翁汤 正丁醇提取物 pH信号通路 PHR2基因 RIM101基因 白念珠菌 黏附 

分 类 号：R285.5[医药卫生—中药学]