摘要
该文基于pH信号通路探讨白头翁汤正丁醇提取物(butyl alcohol extract of Baitouweng Decoction,BAEB)对白念珠菌生物膜的影响。以扫描电镜观察pH突变株生物膜形态结构; CLSM测量pH突变株生物膜厚度;酶标仪检测pH突变株生物膜活性;流式细胞仪检测pH突变株生物膜损伤; qRT-PCR法检测pH突变株生物膜相关基因的表达。结果显示,对生物膜结构,PHR1基因缺失造成结构缺陷,PHR1回补则基质较多; BAEB干预对该2株无明显影响。RIM101基因缺失或回补无明显结构损伤,但BAEB干预后,这2种菌株生物膜均被明显抑制。对生物膜厚度,PHR1基因缺失或PHR1回补的厚度降低;BAEB干预后,该2株厚度变化不明显。而RIM101基因缺失或回补对生物膜厚度影响不大;加入BAEB后,该2株生物膜厚度明显变薄。对生物膜活性,PHR1基因缺失、PHR1回补和RIM101基因缺失均造成活性降低,RIM101回补无明显变化;BAEB干预后,PHR1基因缺失、PHR1回补、RIM101缺失与RIM101回补的生物膜活性与干预前相比显著降低。对生物膜损伤程度,PHR1基因缺失、PHR1回补和RIM101基因缺失、RIM101回补均出现不同程度损伤;加入BAEB干预后,PHR1缺失或PHR1回补的损伤率差异并不大,但RIM101缺失或RIM101回补的损伤率显著升高。对基因表达水平,除HSP90基因上调外,PHR1缺失、PHR1回补、RIM101缺失、RIM101回补的ALS3,SUN41,HWP1,UME6和PGA10基因均出现了不同倍数的下调趋势。该研究表明,pH信号通路中PHR1和RIM101基因突变能提高菌株对于抗真菌药物BAEB的敏感性,从而抑制白念珠菌生物膜形成及相关基因的表达。
This study aimed to investigate the effect of butyl alcohol extract of Baitouweng Decoction( BAEB) on Candida albicans biofilms based on pH signal pathway. The morphology of biofilms of the pH mutants was observed by scanning electron microscope. The biofilm thickness of the pH mutants was measured by CLSM. The biofilm activity of the pH mutants was analyzed by microplate reader.The biofilm damage of the pH mutants was detected by flow cytometry. The expression of pH mutant biofilm-related genes was detected by qRT-PCR. The results showed that the deletion of PHR1 gene resulted in the defect of biofilm,but there were more substrates for PHR1 complementation. BAEB had no significant effect on the two strains. RIM101 gene deletion or complementation did not cause significant structural damage,but after BAEB treatment,the biofilms of both strains were significantly inhibited. For the biofilm thickness,PHR1 deletion or complementation caused the thickness to decrease,after BAEB treatment,the thickness of the two strains did not change significantly. However,RIM101 gene deletion or complementation had little effect on the thickness,and the thickness of the two strains became thinner after adding BAEB. For biofilm activity,PHR1 deletion or complementation and RIM101 deletion resulted in decreased activity,RIM101 complementation did not change significantly;BAEB significantly inhibited biofilm activity of PHR1 deletion,PHR1 complemetation,RIM101 deletion and RIM101 complemetation strains. For the biofilm damage,PHR1 gene deletion or complementation,RIM101 gene deletion or complementation all showed different degrees of damage;after adding BAEB,the damage rate of PHR1 deletion or complementation was not significantly different,but the damage rate of RIM101 deletion or complementation was significantly increased. Except to the up-regulation of HSP90 gene expression,ALS3,SUN41,HWP1,UME6 and PGA10 genes of PHR1 deletion,PHR1 complementation,RIM101 deletion,and RIM101 complementation strains showed a downward expression tren
作者
汪云霞
马克龙
王艳
吴大强
邵菁
汪天明
汪长中
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)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2019年第2期350-356,共7页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81573725
81774034)
安徽省教育厅科研重点项目(KJ2015A030)
