Pos-graduation in Biophotonic, University Nove de Julho - Uninove, São Paulo, São Paulo, Brazil.
Department of Science and Technology, Institute of Science and Technology, Federal University of São Paulo - UNIFESP, Rua Talim, no. 330 - Vila Nair, CEP: 12231-280, São José dos Campos, São Paulo, Brazil.
Inflammation. 2021 Aug;44(4):1643-1661. doi: 10.1007/s10753-021-01448-5. Epub 2021 Mar 17.
The present study was aimed to investigate the phototherapy effect with low-level laser on human bronchial epithelial cells activated by cigarette smoke extract (CSE). Phototherapy has been reported to actuate positively for controlling the generation/release of anti-inflammatory and pro-inflammatory mediators from different cellular type activated by distinct stimuli. It is not known whether the IL-8 and IL-10 release from CSE-stimulated human bronchial epithelium (BEAS) cells can be influenced by phototherapy. Human bronchial epithelial cell (BEAS) line was cultured in a medium with CSE and irradiated (660 nm) at 9 J. Apoptosis index was standardized with Annexin V and the cellular viability was evaluated by MTT. IL-8, IL-10, cAMP, and NF-κB were measured by ELISA as well as the Sp1, JNK, ERK1/2, and p38MAPK. Phototherapy effect was studied in the presence of mithramycin or the inhibitors of JNK or ERK. The IL-8, cAMP, NF-κB, JNK, p38, and ERK1/2 were downregulated by phototherapy. Both the JNK and the ERK inhibitors potentiated the phototherapy effect on IL-8 as well as on cAMP secretion from BEAS. On the contrary, IL-10 and Sp1 were upregulated by phototherapy. The mithramycin blocked the phototherapy effect on IL-10. The results suggest that phototherapy has a dual effect on BEAS cells because it downregulates the IL-8 secretion by interfering with CSE-mediated signaling pathways, and oppositely upregulates the IL-10 secretion through of Sp1 transcription factor. The manuscript provides evidence that the phototherapy can interfere with MAPK signaling via cAMP in order to attenuate the IL-8 secretion from CSE-stimulated BEAS. In addition, the present study showed that phototherapy effect is driven to downregulation of the both the IL-8 and the ROS secretion and at the same time the upregulation of IL-10 secretion. Besides it, the increase of Sp-1 transcription factor was crucial for laser effect in upregulating the IL-10 secretion. The dexamethasone corticoid produces a significant inhibitory effect on IL-8 as well as ROS secretion, but on the other hand, the corticoid blocked the IL-10 secretion. Taking it into consideration, it is reasonable to suggest that the beneficial effect of laser therapy on lung diseases involves its action on unbalance between pro-inflammatory and anti-inflammatory mediators secreted by human bronchial epithelial cells through different signaling pathway.
本研究旨在探讨低强度激光对香烟烟雾提取物(CSE)激活的人支气管上皮细胞的光疗作用。光疗已被报道可积极控制不同细胞类型在不同刺激下产生/释放抗炎和促炎介质。目前尚不清楚 CSE 刺激的人支气管上皮细胞(BEAS)细胞释放的 IL-8 和 IL-10 是否可以受到光疗的影响。将人支气管上皮细胞(BEAS)系培养在含有 CSE 的培养基中,并以 9 J 的能量进行(660nm)照射。用 Annexin V 使凋亡指数标准化,并通过 MTT 评估细胞活力。通过 ELISA 测量 IL-8、IL-10、cAMP 和 NF-κB,以及 Sp1、JNK、ERK1/2 和 p38MAPK。在米托蒽醌或 JNK 或 ERK 抑制剂存在的情况下研究光疗效果。光疗可下调 IL-8、cAMP、NF-κB、JNK、p38 和 ERK1/2。JNK 和 ERK 抑制剂均增强了光疗对 BEAS 细胞中 IL-8 和 cAMP 分泌的作用。相反,IL-10 和 Sp1 被光疗上调。米托蒽醌阻断了光疗对 IL-10 的作用。结果表明,光疗对 BEAS 细胞具有双重作用,因为它通过干扰 CSE 介导的信号通路下调 IL-8 的分泌,而通过 Sp1 转录因子上调 IL-10 的分泌。该研究提供了证据表明,光疗可以通过 cAMP 干扰 MAPK 信号传导,以减轻 CSE 刺激的 BEAS 细胞中 IL-8 的分泌。此外,本研究表明,光疗效果是通过下调 IL-8 和 ROS 分泌,同时上调 IL-10 分泌来驱动的。此外,Sp1 转录因子的增加对于激光上调 IL-10 分泌至关重要。皮质类固醇地塞米松对 IL-8 和 ROS 分泌具有显著的抑制作用,但另一方面,皮质类固醇阻断了 IL-10 的分泌。考虑到这一点,有理由认为激光治疗对肺部疾病的有益作用涉及到它通过不同的信号通路作用于人支气管上皮细胞分泌的促炎和抗炎介质之间的失衡。