Huang Xiaoxue, Zheng Dongming, Yong Jiangyan, Li Yan
West China School of Public Health and West China Fourth Hospital, Sichuan University, Sichuan 610041, PR China.
College of Medical Technology, Chengdu University of Traditional Chinese Medicine, Sichuan 611137, PR China.
J Med Microbiol. 2022 Jun;71(6). doi: 10.1099/jmm.0.001542.
The emergence of resistance to fluconazole in has made the clinical treatment of this microbe difficult. A potential strategy to address this problem involves diminishing fungal resistance to antimicrobial drugs. Berberine hydrochloride (BH), the primary active ingredient of the traditional Chinese medicine (TCM) Coptis, inhibits the growth of fluconazole-resistant through its action on the high-osmolarity glycerol mitogen-activated protein kinase (HOG-MAPK) pathway. To examine the effect of BH on the HOG-MAPK pathway to assess the potential molecular mechanism by which BH inhibits fluconazole-resistant . The minimum inhibitory concentration (MIC) of BH to fluconazole-resistant was measured using the broth microdilution approach to determine the concentration of effective drug intervention. Changes in physiological functions regulated by the HOG-MAPK pathway in response to BH treatment were measured, as well as the expression of central signalling pathway genes and key downstream factors by qRT-PCR and Western blotting, respectively. BH inhibited fluconazole-resistant and the sensitivity to fluconazole increased after BH treatment. At a concentration of 256 and 64 μg ml BH may affect key downstream factors that regulate several physiological functions of by upregulating the core genes expression of , , , and in the HOG-MAPK pathway. Upregulation of , the key gene for glycerol synthesis, increased cell osmotic pressure. BH treatment increased the accumulation of reactive oxygen species by upregulating the expression of the key respiratory metabolism gene and downregulating the expression of the superoxide dismutase gene . Furthermore, downregulation of mycelial-specific hindered the morphological transformation of and inhibition of the chitin synthase gene and the β-(1,3) glucan synthase gene impaired cytoderm integrity. BH affects multiple target genes in diminishing the resistance of strains to fluconazole. This effect may be related to the action of BH on the HOG-MAPK pathway.
对氟康唑产生耐药性使得这种微生物的临床治疗变得困难。解决这一问题的潜在策略包括降低真菌对抗菌药物的耐药性。盐酸小檗碱（BH）是中药黄连的主要活性成分，通过作用于高渗甘油丝裂原活化蛋白激酶（HOG-MAPK）途径抑制耐氟康唑菌株的生长。为了研究BH对HOG-MAPK途径的影响，以评估BH抑制耐氟康唑菌株的潜在分子机制。采用肉汤微量稀释法测定BH对耐氟康唑菌株的最低抑菌浓度（MIC），以确定有效药物干预浓度。测定了HOG-MAPK途径调控的生理功能在BH处理后的变化，以及分别通过qRT-PCR和蛋白质免疫印迹法检测中心信号通路基因和关键下游因子的表达。BH抑制了耐氟康唑菌株，且BH处理后对氟康唑的敏感性增加。在浓度为256和64μg/ml时，BH可能通过上调HOG-MAPK途径中、、、和的核心基因表达来影响调控菌株多种生理功能的关键下游因子。甘油合成关键基因的上调增加了细胞渗透压。BH处理通过上调关键呼吸代谢基因的表达和下调超氧化物歧化酶基因的表达增加了活性氧的积累。此外，菌丝特异性的下调阻碍了菌株的形态转变，几丁质合酶基因和β-(1,3)葡聚糖合酶基因的抑制损害了细胞壁完整性。BH在降低菌株对氟康唑耐药性方面影响多个靶基因。这种作用可能与BH对HOG-MAPK途径的作用有关。
