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Expression and role of aryl hydrocarbon receptor in Aspergillus fumigatus keratitis

International Journal of Ophthalmology(English edition) 2020年第2期13卷 199-205页

作者： Li Zhang Nan Jiang Gui-Qiu Zhao Xu-Dong Peng Guo-Qiang Zhu Wei Jiang Jing-Jing Ma Department of Ophthalmology the Affiliated Hospital of Qingdao UniversityQingdao 266003Shandong ProvinceChina

●AIM:To observe the expression and role of aryl hydrocarbon receptor(Ah R)in the immune response of mouse cornea infected with Aspergillus fumigatus(***).●METHODS:Murine models of *** keratitis were established by scraping the central epithelium of mouse cornea,daubing *** on the cornea and covering with a contact *** mice were randomly divided into the control group and the ***-infected(A.F.)group for 1,3 and 5 d respectively,which corneas were daily monitored by a slit lamp microscope and the clinical scores were also recorded timely after *** this study,immunofluorescence staining was used to detect the expression and localization of Ah R in mouse corneas,and the m RNA and protein of Ah R were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western *** addition,mouse peritoneal macrophages were stimulated by *** with or without the pretreatment of Ah R antagonist CH223191 and Ah R agonist FICZ,and the tumor necrosis factor alpha(TNF-α),inducible nitric oxide synthase(i NOS),interleukin-10(IL-10)and Arg-1 m RNA were detected by RT-PCR.●RESULTS:According to the results of the slit light photography,it was clearly indicated that the corneal inflammation were the most severe and the clinical score became the highest as well on the 3 rd day after the infection of *** with the control group,the expression of Ah R in the corneal epithelial cells infected with *** was significantly increased detected by immunofluorescence *** R mainly expressed in the nucleus and cytoplasm of corneal epithelial *** with the transcriptional level of Ah R m RNA,the expression level of Ah R protein reached the peak on the 3 rd day after infection which was detected by Western ***,RT-PCR showed that CH223191 up-regulated the expression of TNF-αand i NOS and down-regulated the expression of IL-10 and Arg-1 in peritoneal macrophages;inversely,FICZ reduce

关键词： aryl hydrocarbon receptor keratitis Aspergillus fumigatus innate immune response

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Disparate expression of autophagy in corneas of C57BL/6 mice and BALB/c mice after Aspergillus fumigatus infection

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International Journal of Ophthalmology(English edition) 2019年第5期12卷 705-710页

作者： Chen-Yu Li Cui Li Hui Li Gui-Qiu Zhao Jing Lin Qian Wang Xu-Dong Peng Qiang Xu Guo-Qiang Zhu Jia-Qian Jiang Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM: To determine the disparate expression of autophagy in the Aspergillus fumigatus(A. fumigatus) keratitis between susceptible C57 BL/6 mice and resistant BALB/c ***: C57 BL/6 and BALB/c mice were used to establish fungal keratitis models. Disease severity and inflammatory response were observed by slit lamp microscopy in A. fumigatus-infected corneas of C57 BL/6 and BALB/c mice at 1, 3 and 5 d. Hematoxylin-eosin(H&E) staining was used to detect pathological changes of corneas. The expression of autophagy-related proteins Beclin-1, LC3, SQSTM1/p62, and LAMP-1 was assessed by Western blot in C57 BL/6 and BALB/c mice at 1, 3 and 5 d post infection(p.i.). Immunofluorescent staining was used to test the expression of LC3 in corneas after A. fumigatus ***: Keratitis severity was higher in C57 BL/6 mice versus BALB/c mice at 1, 3 and 5 d p.i. H&E staining showed that the number of inflammatory cells was larger and the severity of ulcer was higher in C57 BL/6 mice than in BALB/c mice after stimulation with A. fumigatus. Higher expression of LAMP-1, Beclin-1, and LC3 was shown in C57 BL/6 mice corneas than in BALB/c mice corneas at 1, 3 and 5 d p.i., while the expression of p62 was lower in C57 BL/6 mice. The fluorescence of LC3 was significantly increased in corneas of C57 BL/6 mice compared with BALB/c mice after A. fumigatus ***: The expression of autophagy is higher in corneas of C57 BL/6 mice than in BALB/c mice afterA. fumigatus infection. Autophagy may be positively correlated with keratitis severity and pathological changes.

关键词： fungal keratitis Aspergillus fumigatus autophagy

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Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis

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International Journal of Ophthalmology(English edition) 2019年第5期12卷 711-716页

作者： Qiang Xu Li-Ting Hu Qian Wang Jing Lin Nan Jiang Cui Li Gui-Qiu Zhao Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM: To investigate the expression of macrophage migration inhibitory factor(MIF) and detect its role in the innate immune response of fungal keratitis(FK). METHODS: We collected the paraffin-embedded cornea tissues from 10 FK and 6 ocular trauma patients to explore the MIF expression by immunohistochemistry. Then we cultured telomease-immortalized human corneal epithelial cells(THCEs), stimulated by the hyphae suspension of Aspergillus fumigatus(A. fumigatus) to detect the change of MIF with or without the pretreatment of MIF inhibitor [4-Iodo-6-phenylpyrimidine(4-IPP)] by real-time polymerase chain reaction(PCR). The protein level of MIF was also tested by immunohistochemistry, and the level of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) mRNA were compared between normal, hyphae stimulated and 4-IPP pretreated groups by real-time PCR to study the influence of MIF on the expression of TNF-α and IL-6. Corneal severity of rats' FK models was documented by clinical scores, and real-time PCR. Western blot and immunohistochemistry were used to test the expression of MIF, TNF-α and IL-6 in rats' ***: In the corneas of FK patients, there was much stronger expression of MIF than that in the normal group showed by immunohistochemistry. In cultured THCEs stimulated by A. fumigatus, the expression of MIF became stronger in both immunohistochemistry and PCR at 16, 24, 32 and 48 h post infection(p.i.; Pfumigatus keratitis, the relative mRNA and protein level of MIF increased than thosein the normal group by PCR(at 1 d: P<0.01, 3 d: P<0.01, 5 d: P<0.01), Western blot and immunohistochemistry. After blocked MIF with 4-IPP, the clinical outcomes of rat keratitis showed markedly reduced inflammatory response(P<0.01), with TNF-α and IL-6 decre

关键词： macrophage migration inhibitory factor fungal keratitis innate immune A. fumigatus corneal epithelial cells rats

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Inhibition of LOX-1 alleviates the proinflammatory effects of high-mobility group box 1 in Aspergillus fumigatus keratitis

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International Journal of Ophthalmology(English edition) 2019年第6期12卷 898-903页

作者： Jia-Qian Jiang Cui Li Cong-Xian Cui Yu-Na Ma Gui-Qiu Zhao Xu-Dong Peng Qiang Xu Qian Wang Guo-Qiang Zhu Chen-Yu Li Department of Ophthalmology the Affiliated Hospital of Qingdao University

AIM: To investigate the inflammatory amplification effect of high-mobility group box 1(HMGB1) in Aspergillus fumigatus(A. fumigatus) keratitis and the relationship between lectin-like oxidized low-density lipoprotein receptor 1(LOX-1) and HMGB1 in keratitis immune ***: Phosphate buffer saline(PBS), and Boxb were injected into BALB/c mice subconjunctivally before the corneas were infected with A. fumigatus. RAW264.7 macrophages and neutrophils were pretreated with PBS and Boxb to determine the HMGB1 inflammatory amplification effects. Abdominal cavity extracted macrophages were pretreated with Boxb and Poly(I)(a LOX-1 inhibitor) before A. fumigatus hyphae stimulation to prove the the relationship between the two molecules. LOX-1, interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), macrophage inflammatory protein-2(MIP-2) and IL-10 were assessed by polymerase chain reaction and Western ***: Pretreatment with Boxb exacerbated corneal inflammation. In macrophages and neutrophils, A. fumigatus induced LOX-1, IL-1β, TNF-α and MIP-2 expression in Boxb group was higher than those in PBS group. Poly(I) treatments before infection alleviated the proinflammatory effects of Boxb in abdominal cavity extracted macrophages. Pretreatment with Boxb did not influence Dectin-1 mRNA levels in macrophages and ***: In fungal keratitis, HMGB1 is a proinflammatory factor in the first line of immune response. HMGB1 mainly stimulates neutrophils and macrophages to produce inflammatory cytokines and chemokines during the immune response. LOX-1 participates in HMGB1 induced inflammatory exacerbation in A. fumigatus keratitis.

关键词： Aspergillus fumigatus keratitis high-mobility group box 1 LOX-1

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Role of the IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to Aspergillus fumigatus infection

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International Journal of Ophthalmology(English edition) 2019年第4期12卷 549-556页

作者： Jia You Jing Lin Yi-Fan Zhou Xu-Dong Peng Hong He Cui Li Guo-Qiang Zhu Xue-Qi Zhao Gui-Qiu Zhao Department of Ophthalmology the Affiliated Hospital of Qingdao University Department of Clinical Laboratory the Affiliated Hospital of Qingdao University

AIM: To investigate the expression of interleukin(IL)-33 in the cornea and human corneal epithelial cells(HCECs) exposed to Aspergillus fumigatus(A. fumigatus), and to determine the function of IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to A. fumigatus ***: The mRNA and protein expression of IL-33 in HCECs and mice corneas were examined by quantitative real-time reverse transcription polymerase chain reaction(q RT-PCR) and Western blot analysis, respectively. IL-33 expression was also detected in cornea samples from healthy donors and patients with fungal keratitis with immunohistochemistry. The cultured HCECs were treated with inactive A. fumigatus hyphae at various concentrations with or without recombinant human IL-33 protein, soluble recombinant ST2 protein, specific ST2 neutralizing antibody, or the mitogen-activated protein kinase(MAPK) p38 inhibitor SB203580 for evaluation of the expression and activation of IL-33/ST2/p38 signaling in the regulation of proinflammatory cytokines. The production levels of IL-6 and IL-1β were determined by qR T-PCR and enzymelinked immunosorbent assay(ELISA). The proliferation of HCECs was determined by a Cell Counting Kit-8(CCK8) assay and cell ***: IL-33 expression levels increased in the corneal tissues of patients with fungal keratitis and in mice corneas of experimental A. fumigatus infection,as well as in HCECs with infection of A. fumigatus. A. fumigatus strongly stimulated HCECs-generated proinflammatory cytokine(IL-6 and IL-1β) production at both the mRNA and protein levels. This production of proinflammatory mediators stimulated by A. fumigatus was further stimulated by IL-33 and was prevented by soluble ST2 protein or ST2 neutralizing antibody. Moreover, IL-33 naturally promoted the p38 phosphorylation induced by A. fumigatus, which was suppressed by soluble ST2 protein. The MAPK p38 inhibitor SB203580 also inhibited the A. fumigatus-induced proinflammato

关键词： ST2 interleukin 33 Aspergillus fumigatus cornea p38 mitogen-activated protein kinase proliferation

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Proteomic Differences between Azole-Susceptible and -Resistant <i>Aspergillus fumigatus</i>Strains

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Advances in Microbiology 2018年第1期8卷 77-99页

作者： Edith Vermeulen Sebastien Carpentier Olaf Kniemeyer Machteld Sillen Johan Maertens Katrien Lagrou Department of Microbiology and Immunology University of Leuven Leuven Belgium Facility for Systems Biology Based Mass Spectrometry University of Leuven Leuven Belgium Leibniz Institute for Natural Product Research and Infection Biology—Hans Knoell Institute (HKI) Molecular and Applied Microbiology Jena Germany Department of Microbiology and Immunology and UZ Leuven Clinical Department of Hematology University of Leuven Leuven Belgium Department of Microbiology and Immunology and UZ Leuven Clinical Department of Laboratory Medicine and National Reference Center for Mycosis University of Leuven Leuven Belgium

Background: Azole-resistance is increasingly reported in Aspergillus fumigatus infections. It remains challenging to rapidly assess antifungal susceptibility to initiate the appropriate therapy. The aim of this study was to map the proteomic differences of azole-susceptible and -resistant strains. Methods: Proteomic studies were performed with ultra-performance liquid chromatography tandem mass-spectrometry (UPLC-MS/MS). Results: UPLC-MS/MS detected 7899 peptides, of which 1792 peptides had a significantly different abundance (p A. fumigatus resistance. Conclusions: UPLC-MS/MS provided proof-of-concept that the proteome of azole-resistant A. fumigatus is diverse enough to serve as a diagnostic tool.

关键词： Aspergillus fumigatus Triazole Resistance Proteomic Mass Spectrometry

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海洋真菌Aspergillus fumigatus次生代谢产物研究

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天然产物研究与开发 2016年第12期28卷 1857-1863页

作者：毛瑞华吴文明方伟阮汉利华中科技大学同济医学院药学院武汉430030 湖北省农业科学院国家生物农药工程研究中心武汉430070

对来源于腔节藻属海藻Coelarthrumboergesenii（采自中国南海西沙群岛）的一株海洋内生真菌Aspergillusfumigatus的发酵液进行化学成分的分离及鉴定。采用硅胶柱色谱、凝胶柱色谱和高压液相色谱等分离手段，从乙酸乙酯萃取部分中分离得... 详细信息

对来源于腔节藻属海藻Coelarthrumboergesenii（采自中国南海西沙群岛）的一株海洋内生真菌Aspergillusfumigatus的发酵液进行化学成分的分离及鉴定。采用硅胶柱色谱、凝胶柱色谱和高压液相色谱等分离手段，从乙酸乙酯萃取部分中分离得到16个次级代谢产物（1-16），经核磁共振、质谱等现代波谱技术鉴定了它们的结构，分别为gliotoxin （ 1 ）. bis （dethio） bis （methylsulfanyl） gliotoxin （ 2 ）, fumiquinazoline C （ 3 ）. （-） -chaetomium （ 4 ）.verruculogen（5） .fumitremorgin C（6） .brevianamide F（7） .fumigaclavine C（8） .decanedioic acid,2-methylene（9） .decanedioic acid, 2-methylene-, 1-methyl ester （ 10 ）. 2-undecenedioic acid （ 11 ）. 2-undecenedioic acid, 1-methyl ester （ 12 ）. helvolic acid （ 13 ）. di-2-ethyl hexyl phthalate （ 14 ）. physcion （ 15 ）. emodin （16）,其中化合物9、10和12为新天然产物。

关键词：海洋真菌内生真菌 Aspergillus fumigatus 代谢产物生物碱

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海洋真菌烟曲霉Aspergillus fumigatus YK-7次级代谢产物的分离鉴定及抗肿瘤活性

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中国药物化学杂志 2017年第5期27卷 382-387页

作者：王宇李占林白皎陈再兴华会明刘涛中国医科大学药学院天然药物化学教研室辽宁沈阳110122 沈阳药科大学基于靶点的药物设计与研究教育部重点实验室辽宁沈阳110016 沈阳药科大学中药学院辽宁沈阳110016 中国医科大学药学院中心实验室辽宁沈阳110122

目的研究海洋真菌烟曲霉Aspergillus fumigatus YK-7次级代谢产物,以期获得活性先导化合物。方法采用硅胶柱色谱、ODS柱色谱、凝胶柱色谱、高效液相色谱等方法进行分离纯化,通过理化性质和波谱数据分析鉴定化合物的结构。采用台盼蓝法和... 详细信息

目的研究海洋真菌烟曲霉Aspergillus fumigatus YK-7次级代谢产物,以期获得活性先导化合物。方法采用硅胶柱色谱、ODS柱色谱、凝胶柱色谱、高效液相色谱等方法进行分离纯化,通过理化性质和波谱数据分析鉴定化合物的结构。采用台盼蓝法和MTT法进行抗肿瘤活性测试。结果从烟曲霉的真菌发酵液的乙酸乙酯萃取物和菌丝体的丙酮提取物中分离得到10个生物碱类化合物,分别鉴定为chaetominine(1)、(-)-11-epi-chaetominine(2)、fumiquinazoline A(3)、fumiquinazoline B(4)、fumiquinazoline C(5)、fumiquinazoline F(6)、fumiquinazoline G(7)、fumiquinazoline J(8)、bis-N-norgliovictin(9)、2-methylthio-cyclo(Phe-Ser)(10)。体外抗肿瘤活性测试结果显示,化合物3、6、7、8对人单核细胞白血病细胞(U937)的IC50值分别为87.1、16.4、33.3、8.6μmol·L^(-1);化合物8对人前列腺癌细胞(PC-3)的IC50值为58.5μmol·L^(-1)。结论化合物1、2为首次从海洋来源烟曲霉中分离得到。化合物3、6、7、8对选定的人肿瘤细胞U937和PC-3具有一定的体外抗增殖活性。

关键词：海洋真菌 Aspergillus fumigatus 次级代谢产物抗肿瘤活性

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海洋真菌Aspergillus fumigatus YK-7次级代谢产物的研究

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化学与生物工程 2020年第3期37卷 25-28,41页

作者：钱诗筠刘涛华会明王宇中国医科大学药学院辽宁沈阳110122 沈阳药科大学中药学院辽宁沈阳110016

对一株海洋真菌Aspergillus fumigatus YK-7的次级代谢产物进行了分离纯化、结构鉴定和抗肿瘤活性测定。利用硅胶柱色谱、ODS柱色谱、凝胶柱色谱、高效液相色谱等方法从该菌株发酵液的乙酸乙酯萃取物和菌丝体的丙酮提取物中分离得到10... 详细信息

对一株海洋真菌Aspergillus fumigatus YK-7的次级代谢产物进行了分离纯化、结构鉴定和抗肿瘤活性测定。利用硅胶柱色谱、ODS柱色谱、凝胶柱色谱、高效液相色谱等方法从该菌株发酵液的乙酸乙酯萃取物和菌丝体的丙酮提取物中分离得到10个化合物。通过核磁与质谱数据分析结合理化常数鉴定化合物的结构,分别为:5α,8α-过氧-(22E,22R)-麦角甾-6,22-二烯-3β-醇(Ⅰ)、(22E,24R)-麦角甾-8,22-二烯-3β,5α,6β,7α-四醇(Ⅱ)、环(脯-色)二肽(Ⅲ)、环(苯丙-丝)二肽(Ⅳ)、4,8-二羟基-α-四氢萘酮(Ⅴ)、苔黑酚(Ⅵ)、苔黑酚羧酸(Ⅶ)、原儿茶酸(Ⅷ)、2-acetylamino-3,5,14-trihydroxyicos-6-en-4-olide(Ⅸ)、1-单亚油酸甘油酯(Ⅹ)。其中,化合物Ⅸ为新天然产物,化合物Ⅱ、Ⅶ、Ⅷ、Ⅹ从曲霉属真菌中首次分离得到,化合物Ⅳ、Ⅵ从该种真菌中首次分离得到。化合物Ⅳ、Ⅴ、Ⅸ对选定的人肿瘤细胞具有一定的体外抗增殖活性。

关键词：海洋真菌 Aspergillus fumigatus 次级代谢产物抗肿瘤活性

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海洋来源真菌烟曲霉(Aspergillus fumigatus)次级代谢产物研究(Ⅱ)

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青岛科技大学学报（自然科学版） 2007年第5期28卷 390-393页

作者：赵文英朱庆书顾谦群青岛科技大学化工学院山东青岛266042 中国海洋大学海洋药物教育部重点实验室山东青岛266042

对一株海洋来源的微生物（Aspergillus fumigatus.）中的次级代谢产物进行了研究,采用硅胶柱色谱方法进行分离纯化,根据理化性质和光谱数据鉴定了化合物的结构,分别为环-（丙-缬）（1）,环-（色-脯）（2）,N-[2-（4-羟基苯基）乙基]乙... 详细信息

对一株海洋来源的微生物（Aspergillus fumigatus.）中的次级代谢产物进行了研究,采用硅胶柱色谱方法进行分离纯化,根据理化性质和光谱数据鉴定了化合物的结构,分别为环-（丙-缬）（1）,环-（色-脯）（2）,N-[2-（4-羟基苯基）乙基]乙酰胺（3）,对羟基苯乙酰胺（4）,4,8-二羟基-1-四氢萘醌（5）,3,4-二羟基苯甲酸甲酯（6）,麦角甾醇（7）和过氧化麦角甾醇（8）。化合物1~8均首次从该菌株中分离。

关键词：海洋来源真菌 Aspergillus fumigatus 次级代谢成份

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