Department of Ophthalmology, St. Vincent Hospital, College of Medicine, The Catholic University of Korea , Seoul , Korea .
Curr Eye Res. 2013 Dec;38(12):1207-13. doi: 10.3109/02713683.2013.811262. Epub 2013 Jul 10.
PURPOSE: To determine the impact of β-1,3-glucan isolated from Euglena gracilis on corneal epithelial cell migration and on wound healing in a rat alkali burn model. METHODS: Immortalized human corneal epithelial cells (HCECs) were cultured in media with 50, 100 and 200 μg/mL laminarin (β-1,3- and β-1,6-glucans), β-1,3-glucan and hyaluronic acid (HA)-conjugated β-1,3-glucan; Dulbecco's modified Eagle's medium (DMEM)/nutrient mixture F-12 (negative control) and serum containing DMEM/nutrient mixture F-12 (positive control). Migration assays were conducted via the manual scraping of HCECs. Next, alkali injuries were induced using 1 N NaOH in 40 eyes of 40 Sprague-Dawley male rats. The injury-only group (n = 10) received topical phosphate-buffered saline four times a day for 2 d. The study groups received 200 μg/mL topical laminarin (n = 10), β-1,3-glucan (n = 10) and β-1,3-glucan HA (n = 10). Using slit-lamp biomicroscopy, injured corneas were evaluated in terms of re-epithelialization and opacity, and tissue sections were histologically analyzed. RESULTS: Migration assay rates were enhanced as laminarin and β-1,3-glucan increased, compared to negative control cells (all p < 0.05). In the comparison between β-1,3-glucan and its HA conjugate form, β-1,3-glucan-HA showed more enhanced migration rate than β-1,3-glucan (p < 0.05). In rat alkali burn model, wound-healing ratio was greatest in β-1,3-glucan-HA groups (96.0 ± 4.1%), followed by β-1,3-glucan (86.0 ± 6.5%), laminarin (67.0 ± 7.5%) and injury-only group (54.0 ± 6.5%) (p < 0.0001; ANOVA). The opacity score was also lowest in β-1,3-glucan-HA groups (3.0 ± 0.75), followed by β-1,3-glucan (3.4 ± 0.5), laminarin (3.7 ± 0.8) and injury-only group (4.7 ± 0.46) (p < 0.0001; ANOVA) Histologically, relatively fewer polymorphonuclear leukocytes infiltrated the corneal stroma in the β-1,3-glucan and β-1,3-glucan-HA groups, compared to the injury-only group. CONCLUSIONS: β-1,3-Glucan, particularly when conjugated with HA, promoted epithelial wound healing in vitro and suppressed the acute inflammatory reaction in corneal alkali burns.
目的：确定从绿眼虫中分离出的β-1,3-葡聚糖对角膜上皮细胞迁移的影响，并在大鼠碱烧伤模型中观察其对伤口愈合的影响。
方法：将永生化人角膜上皮细胞（HCEC）在含有 50、100 和 200μg/mL 昆布多糖（β-1,3-和β-1,6-葡聚糖）、β-1,3-葡聚糖和透明质酸（HA）结合的β-1,3-葡聚糖、无血清 DMEM/营养混合物 F-12（阴性对照）和含血清的 DMEM/营养混合物 F-12（阳性对照）的培养基中进行培养。通过手动刮除 HCEC 进行迁移实验。然后，在 40 只 Sprague-Dawley 雄性大鼠的 40 只眼中用 1N NaOH 诱导碱烧伤。仅损伤组（n=10）每天接受 4 次局部磷酸盐缓冲盐水治疗 2 天。研究组接受 200μg/mL 局部昆布多糖（n=10）、β-1,3-葡聚糖（n=10）和β-1,3-葡聚糖-HA（n=10）治疗。使用裂隙灯生物显微镜评估受伤角膜的再上皮化和混浊程度，并对组织切片进行组织学分析。
结果：与阴性对照细胞相比，昆布多糖和β-1,3-葡聚糖均能提高迁移率（均 p<0.05）。在β-1,3-葡聚糖与其 HA 结合形式的比较中，β-1,3-葡聚糖-HA 比β-1,3-葡聚糖表现出更高的迁移率（p<0.05）。在大鼠碱烧伤模型中，β-1,3-葡聚糖-HA 组的愈合比例最大（96.0±4.1%），其次是β-1,3-葡聚糖（86.0±6.5%）、昆布多糖（67.0±7.5%）和仅损伤组（54.0±6.5%）（p<0.0001；方差分析）。β-1,3-葡聚糖-HA 组的混浊评分也最低（3.0±0.75），其次是β-1,3-葡聚糖（3.4±0.5）、昆布多糖（3.7±0.8）和仅损伤组（4.7±0.46）（p<0.0001；方差分析）。组织学上，与仅损伤组相比，β-1,3-葡聚糖和β-1,3-葡聚糖-HA 组角膜基质中浸润的多形核白细胞相对较少。
结论：β-1,3-葡聚糖，特别是与 HA 结合时，可促进体外上皮伤口愈合，并抑制角膜碱烧伤的急性炎症反应。
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