Sibin Melo Katia C, Correia Marcelo H, Svidzinski Terezinha I E, Hernandes Luzmarina
Department of Morphological Sciences, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.
Department of Clinical Analysis and Biomedicine, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.
APMIS. 2018 May;126(5):418-427. doi: 10.1111/apm.12835.
The skin is an important gateway for Fusarium infection in humans. Our hypothesis is that metabolites produced by Fusarium oxysporum should change the barrier structure to permeate the skin. Male Wistar rats received a topical application of a solution (0.05 mg/mL) of Fusarium metabolites. The animals were euthanized 3, 6, 12, 24 h after and the skin was processed for immunostaining by laminin and E-cadherin to investigate whether the Fusarium metabolites can break the barrier of healthy skin. Other techniques were employed: H&E to study the morphology; metalloproteinase-9 (MMP-9), TUNEL, and PCNA immunostaining to evaluate the inflammation, cell death, and proliferation, respectively. There was an inflammatory response mainly centered in the dermis. Qualitatively, the skin of the experimental group showed reduced E-cadherin and laminin immunostaining at 3, 12, and 24 h. Higher intensity staining by TUNEL at 3 h, and PCNA at 6, 12, and 24 h. There was intense MMP-9 activity at 6, 12, and 24 h. None of analyses revealed any changes in the epidermis. It was concluded that the fraction was able to permeate the skin and act selectively in dermis, inducing inflammatory response, increasing MMP-9 immunostaining, inducing apoptosis, and reducing E-cadherin and laminin immunostaining.
皮肤是人类镰刀菌感染的重要途径。我们的假设是尖孢镰刀菌产生的代谢产物会改变屏障结构以穿透皮肤。雄性Wistar大鼠局部应用镰刀菌代谢产物溶液(0.05 mg/mL)。在给药后3、6、12、24小时对动物实施安乐死,并对皮肤进行层粘连蛋白和E-钙黏蛋白免疫染色处理,以研究镰刀菌代谢产物是否会破坏健康皮肤的屏障。还采用了其他技术:苏木精-伊红染色(H&E)来研究形态学;金属蛋白酶-9(MMP-9)、末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL)和增殖细胞核抗原(PCNA)免疫染色,分别用于评估炎症、细胞死亡和增殖情况。主要在真皮层出现了炎症反应。定性分析显示,实验组皮肤在3、12和24小时时E-钙黏蛋白和层粘连蛋白免疫染色减少。TUNEL在3小时时染色强度较高,PCNA在6、12和24小时时染色强度较高。MMP-9在6、12和24小时时活性较强。所有分析均未发现表皮有任何变化。得出的结论是,该组分能够穿透皮肤并在真皮中选择性发挥作用,引发炎症反应,增加MMP-9免疫染色,诱导细胞凋亡,并减少E-钙黏蛋白和层粘连蛋白免疫染色。