伤口愈合
TLR5型
活力测定
TLR2型
兴奋剂
药理学
炎症
抗菌肽
MTT法
促炎细胞因子
体内
体外
医学
化学
分子生物学
免疫学
微生物学
TLR4型
生物
抗菌剂
受体
内科学
生物化学
生物技术
作者
Carolina Lema,Hasna Baidouri,Mingxia Sun,Susanne Pohl,Sharon Cookson,Rachel L. Redfern,Alison M. McDermott
标识
DOI:10.1016/j.jtos.2022.09.003
摘要
In the skin, Lucilia sericata maggot excretions/secretions (ES) accelerate wound healing and limit inflammation. This study aimed to determine whether ES have similar beneficial effects at the ocular surface. Human corneal epithelial cells (HCEC) were cultured with ES and cell viability was determined by the MTT assay. Additionally, mRNA expression of growth factors, antimicrobial peptides (AMPs) and cytokines was assessed by qPCR. ES ability to modulate TLR-induced IL-6 and IL-8 expression was determined by qPCR and ELISA. ES potential to promote corneal healing was evaluated in vitro by a migration assay in HCEC, and in vivo using a mouse model. ES did not impair HCEC viability up to 25 μg/ml. Among the factors evaluated, only hBD-2 was upregulated (2.5-fold) by 1.5 μg/ml ES after 6 hrs (P = 0.04). In HCEC, ES reduced Poly I:C-induced IL-6 and IL-8 mRNA (P ≤ 0.001) and protein (P ≤ 0.0001) expression. A similar effect was observed with Flagellin (TLR5 agonist) but it was less robust for FSL-1 (TLR2/6 agonist) and Pam3CSK4 (TLR1/2 agonist). The greatest in vitro migration effect was observed with 6.2 μg/ml ES after 44 hrs where gap area compared to vehicle was 53.3 ± 3.7% vs. 72.6 ± 5.4% (P = 0.001). In the mouse model, the maximum healing effect was present with 1.5 μg/ml ES after 12 hrs with a wound area of 19.0 ± 2.7% vs. 60.1 ± 21.6% (P = 0.003) or 77% reduction of the wound area compared to the negative control. ES significantly reduce in vitro TLR-induced production of inflammatory cytokines and promote corneal wound healing.
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