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

中国真菌学杂志　 2020, Vol. 15 　Issue (2): 65-71.

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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

张静云¹, 彭靖雯¹, 王琼¹, 高盈¹, 陈万鑫¹, 沈永年¹, Li Dong-mei², 佘晓东¹, 刘维达¹

1. 中国医学科学院皮肤病医院, 南京 210042;
2. Georgetown University, DC USA 20057

收稿日期:2019-09-26 出版日期:2020-04-28 发布日期:2020-04-28
通讯作者: 佘晓东,E-mail:shexd1979@163.com;刘维达,E-mail:liumyco@hotmail.com E-mail:shexd1979@163.com;liumyco@hotmail.com
作者简介:张静云,女(汉族),硕士研究生在读,E-mail:18370959919@163.com
基金资助:
江苏省自然科学基金面上项目（BK20191137）

The role of EGFR in the pathogenesis of vaginal candidiasis

ZHANG Jing-yun¹, PENG Jing-wen¹, WANG Qiong¹, GAO Ying¹, CHEN Wan-xin¹, SHEN Yong-nian¹, LI Dong-mei², SHE Xiao-dong¹, LIU Wei-da¹

1. Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042;
2. Department of Microbiology&Immunology, Georgetown University, Medical Center, Washington DC 20057, USA

Received:2019-09-26 Online:2020-04-28 Published:2020-04-28

摘要/Abstract

摘要：

目的初步探索表皮生长因子受体（epidermal growth factor receptor，EGFR）在阴道念珠菌病发病过程中的作用及分子机制。方法培养阴道上皮细胞（VK2/E6E7细胞），白念珠菌刺激后利用荧光定量PCR检测EGFR及可能相关的免疫通路因子的表达。构建EGFR-siRNA VK2/E6E7细胞模型，与白念珠菌共培养后，分别利用ELLISA及全自动细胞检测仪检测表皮生长因子受体敲除前后阴道上皮细胞分泌细胞因子的变化以及对白念珠菌感染防御能力的改变。构建阴道念珠菌感染小鼠模型，qPCR检测阴道组织表皮生长因子受体及免疫通路因子的表达；并检测EGFR磷酸化抑制剂阻断通路后，阴道组织局部的真菌载量和炎性细胞的变化。结果 qPCR检测显示，EGFR、STAT3、GM-CSF及IL-1β在VK2/E6E7细胞感染白念珠菌后表达升高，且具有统计学意义（P<0.05）；ELLISA检测结果显示，EGFR-siRNA VK2/E6E7细胞感染白念珠菌后GM-CSF、IL-8、 IL-1β、 MIP-3α表达显著下降（P<0.05）；全自动细胞检测仪检测结果显示，EGFR敲除细胞在感染白念珠菌30 h后的防御能力和细胞活性较正常细胞明显降低。qPCR检测显示小鼠阴道感染白念珠菌后EGFR、HER2、STAT3、IL-8表达升高具有统计学意义（P<0.05）；局部应用磷酸化抑制剂阻断EGFR通路后，小鼠阴道灌洗液菌载量较对照组明显增加，且在感染后第7日差异有统计学意义（P<0.05）。结论表皮生长因子受体及其通路因子在阴道念珠菌病发病过程中起到较为重要的作用。

关键词: 表皮生长因子受体, 白念珠菌, 阴道念珠菌病

Abstract:

Objective To investigate the role and molecular mechanism of EGFR in the pathogenesis of vaginal candidiasis. Methods Vaginal epithelial cells (VK2/E6E7 cells) were cultured. After stimulation with Candida albicans, the expression of immune factors in EGFR and possible related immune pathway factors were detected by real-time PCR. The EGFR-siRNA VK2/E6E7 cell model was constructed and co-cultured with Candida albicans. The cytokines secreted by vaginal epithelial cells and the protective ability against Candida albicans infection were detected by ELLISA and automatic cell detector. A mouse model of vaginal Candida infection was constructed. qPCR was used to detect the expression of EGFR and immune pathway factors in vaginal tissues. The changes of fungal load and inflammatory cells in vaginal tissues were detected after EGFR phosphorylation inhibitor blocking pathway. Results qPCR assay showed that EGFR, STAT3, GM-CSF and IL-1β were up-regulated in VK2/E6E7 cells infected with Candida albicans, and were statistically significant (P <0.05). ELLISA assay showed the expressions of GM-CSF, IL-8, IL-1β and MIP-3α were significantly decreased in EGFR-siRNA VK2/E6E7 cells infected with Candida albicans (P<0.05). The results of automatic cell detector showed that the defense capacity and cell activity of EGFR knockout cells after 30 hours' infection of Candida albicans were significantly lower than normal cells. The results of qPCR showed that the expression of EGFR, HER2, STAT3 and IL-8 following the vaginal infection of Candida albicans in the mouse was statistically significant (P<0.05); after local application of phosphorylation inhibitor, the lavage fluid load in the mouse vagina was significantly higher than that of the control group with EGFR pathway being blocked and there was a statistical difference on the 7th day after infection (P<0.05). Conclusion The EGFR and its pathway factors should play an important role in the pathogenesis of vaginal candidiasis.

Key words: EGFR, Candida albicans, vaginal candidiasis

中图分类号:

R519.3

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.

ZHANG Jing-yun, PENG Jing-wen, WANG Qiong, GAO Ying, CHEN Wan-xin, SHEN Yong-nian, LI Dong-mei, SHE Xiao-dong, LIU Wei-da. The role of EGFR in the pathogenesis of vaginal candidiasis[J]. Chinese Journal of Mycology, 2020, 15(2): 65-71.

参考文献

[1] Sobel JD. Recurrent vulvovaginal candidiasis[J].Am J Obstet Gynecol, 2016,214(1):15-21.
[2] Cassone A. Vulvovaginal Candida albicans infections:pathogenesis, immunity and vaccine prospects[J]. BJOG, 2015,122(6):785-794.
[3] 佘晓东, 沈永年, 韩峻松, 等. 用Oligo芯片和小鼠模型研究外阴阴道念珠菌病的发病机制[J].中华皮肤科杂志, 2009, 42:681-684.
[4] Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer[J]. Mol Oncol,2018,12(1):3-20.
[5] Mascia F, Cataisson C, Lee TC, et al. EGFR regulates the expression of keratinocyte-derived granulocyte/macrophage colony-stimulating factor in vitro and in vivo[J]. J Invest Dermatol,2010,130(3):682-693.
[6] Shu DY, Wojciechowski M, Lovicu FJ. ERK1/2-mediated EGFR-signaling is required for TGFβ-induced lens epithelial-mesenchymal transition[J]. Exp Eye Res, 2019,178(1):108-121.
[7] Ray K, Ujvari B, Ramana V, et al. Cross-talk between EGFR and IL-6 drives oncogenic signaling and offers therapeutic opportunities in cancer[J]. Cytokine Growth Factor Rev, 2018,41(1):18-27.
[8] Chattopadhyay S, Veleeparambil M, Poddar D, et al. EGFR kinase activity is required for TLR4 signaling and the septic shock response[J]. EMBO Rep, 2015,16(11):1535-1547.
[9] Veleeparambil M, Poddar D, Abdulkhalek S, et al.Constitutively bound EGFR-mediated tyrosine phosphorylation of TLR9 is required for its ability to signal[J]. J Immunol,2018,200(8):2809-2818.
[10] Zhu W, Phan QT, Boontheung P, Solis NV, et al. EGFR and HER2 receptor kinase signaling mediate epithelial cell invasion by Candida albicans during oropharyngeal infection[J]. Proc Natl Acad Sci U S A,2012, 109:14194-14199.
[11] Wang SH, Wang SC, Chen PC, et al. Induction of cyclooxygenase-2 gene by Candida albicans through EGFR, ERK, and p38 pathways in human urinary epithelium[J]. Med Mycol,2017,55(3):314-322.
[12] Lionakis MS, Lim JK, Lee CC, et al. Organ-specific innate immune responses in a mouse model of invasive candidiasis[J]. J Innate Immun, 2011, 3(2):180-199.

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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

The role of EGFR in the pathogenesis of vaginal candidiasis

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