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盐酸小檗碱对白色念珠菌菌丝细胞壁完整性的影响

[Effect of berberine hydrochloride on cell wall integrity of Candida albicans hypha].

作者信息

Yang Yu, Wang Ya-Dong, Wang Yan, Xu Zhi-Qing, Wu Da-Qiang, Wang Tian-Ming, Shao Jing, Wang Chang-Zhong

机构信息

School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine Hefei 230012, China Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine Hefei 230012, China.

School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine Hefei 230012, China Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine Hefei 230012, China Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine Hefei 230012, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2021 Jan;46(1):155-161. doi: 10.19540/j.cnki.cjcmm.20200907.401.

Abstract

The aim of this paper was to investigate the effect of berberine hydrochloride on the cell wall integrity of Candida albicans hypha. The minimal inhibitory concentration(MIC) of berberine hydrochloride against clinical and standard C. albicans strains was detected by micro liquid-based dilution method; the effect of berberine hydrochloride on the colony formation of C. albicans SC5314 was investigated by spot assay; the effect of berberine hydrochloride on the metabolism of C. albicans SC5314 hypha was checked by XTT reduction assay, and the viability of C. albicans SC5314 hypha was tested by fluorescent staining assay. The effect of berberine hydrochloride on the morphology of C. albicans SC5314 hypha was examined by scanning electron microscope. The changes in the cell wall of C. albicans SC5314 hypha after berberine hydrochloride treatment were detected by transmission electron microscopy. The effect of berberine hydrochloride on β-glucan from C. albicans SC5314 was detected by flow cytometry. The effect of berberine hydrochloride on hypha-specific gene ECE1 and β-glucan synthase genes FKS1 and FKS2 in C. albicans was examined by qRT-PCR. The results showed that berberine hydrochloride showed a strong inhibitory effect on both clinical and standard strains of C. albicans, and the MIC was 64-128 μg·mL~(-1). Spot assay, XTT redunction assay and fluorescent staining assay showed that with the increase of berberine hydrochloride concentration, the viability of C. albicans SC5314 gradually decreased. The transmission electron microscopy scanning assay showed that this compound could cause cell wall damage of C. albicans. The flow cytometry analysis showed the exposure degree of C. albicans β-glucan. The qRT-PCR further showed that berberine hydrochloride could significantly down-regulate hypha-specific gene ECE1 and β-glucan synthase-related gene FKS1 and FKS2. In conclusion, this compound can down-regulate C. albicans and β-glucan synthase-related gene expressions, so as to destroy the cell wall structure of C. albicans, expose β-glucan and damage the integrity of the wall.

摘要

本文旨在研究盐酸小檗碱对白色念珠菌菌丝细胞壁完整性的影响。采用微量液体稀释法检测盐酸小檗碱对临床和标准白色念珠菌菌株的最小抑菌浓度(MIC);采用点样法研究盐酸小檗碱对白色念珠菌SC5314菌落形成的影响;采用XTT还原法检测盐酸小檗碱对白色念珠菌SC5314菌丝代谢的影响,采用荧光染色法检测白色念珠菌SC5314菌丝的活力。通过扫描电子显微镜观察盐酸小檗碱对白色念珠菌SC5314菌丝形态的影响。采用透射电子显微镜检测盐酸小檗碱处理后白色念珠菌SC5314菌丝细胞壁的变化。采用流式细胞术检测盐酸小檗碱对白色念珠菌SC5314β-葡聚糖的影响。采用qRT-PCR检测盐酸小檗碱对白色念珠菌菌丝特异性基因ECE1和β-葡聚糖合酶基因FKS1和FKS2的影响。结果表明,盐酸小檗碱对临床和标准白色念珠菌菌株均有较强的抑制作用,MIC为64 - 128μg·mL⁻¹。点样法、XTT还原法和荧光染色法表明,随着盐酸小檗碱浓度的增加,白色念珠菌SC5314的活力逐渐降低。透射电子显微镜扫描分析表明,该化合物可导致白色念珠菌细胞壁损伤。流式细胞术分析显示了白色念珠菌β-葡聚糖的暴露程度。qRT-PCR进一步表明,盐酸小檗碱可显著下调菌丝特异性基因ECE1和β-葡聚糖合酶相关基因FKS1和FKS2。综上所述,该化合物可下调白色念珠菌及β-葡聚糖合酶相关基因表达,从而破坏白色念珠菌细胞壁结构,暴露β-葡聚糖,损害细胞壁完整性。

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