1400W 对阻断脂多糖诱导小胶质细胞毒性对少突胶质前体细胞的影响。

Effect of 1400W on blocking lipopolysaccharide-induced microglial toxicity to preoligodendrocytes.

机构信息

Shanghai Institute for Pediatric Research, Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.

出版信息

World J Pediatr. 2010 Aug;6(3):249-54. doi: 10.1007/s12519-010-0203-2. Epub 2010 Jun 12.

DOI:10.1007/s12519-010-0203-2

PMID:20549418

Abstract

BACKGROUND

The maternal-fetal infection/inflammation is believed to be the mechanism in the pathogenesis of periventricular leukomalacia (PVL). The activation of microglias (MGs) may contribute to preoligodendroglial damage. The present study was undertaken to explore the effect of N-[3-(aminomethyl) benzyl] acetamidine (1400W), a selective inhibitor of inducible nitric oxide synthase (iNOS), on the blockage of lipopolysaccharide (LPS)-induced microglial toxicity to preoligodendrocytes (preOLs).

METHODS

The co-cultural MGs and preOLs obtained from two-day-old Sprague-Dawley (SD) neonatal rats were divided into three groups: co-culture control group, coculture LPS group, and co-culture LPS plus 1400W group. The concentration of nitric oxide (NO) was measured by nitric acid-deoxidize-colorimetry, the level of peroxynitrite (ONOO(-)) determined by immunocytochemistry, the synthetic level of inducible nitric oxide synthase (iNOS) detected by western blotting, and the apoptotic rate of preOLs assessed by flow cytometry after the co-cultural cells were induced by LPS (100 ng/ml) for 48 hours.

RESULTS

Compared with those in the co-culture control group, the levels of NO (82.27+/-3.41 micromol/L vs. 167.86+/-9.87 micromol/L, P<0.01), ONOO(-) (6.14+/-1.27 vs. 34.38+/-7.75, P<0.01), and iNOS (0.18+/-0.027 vs. 0.79+/-0.068, P<0.01) induced by LPS increased remarkably in the co-culture LPS group, with a higher apoptotic rate of preOLs (6.73+/-1.39% vs. 24.77+/-2.05%, P<0.01). The levels of NO (69.55+/-5.07 micromol/L, P<0.01), ONOO(-) (10.33+/-3.47, P<0.01) and iNOS (0.35+/-0.042, P<0.01) were decreased significantly using 1400W at a dose of 10 micromol/L in the co-culture LPS plus 1400W group, and the apoptotic rate of preOLs (11.80+/-2.06% vs. 24.77+/-2.05%, P<0.01) also decreased compared with the co-culture LPS group.

CONCLUSION

1400W can block effectively the LPS-induced microglial toxicity to preOLs by inhibiting iNOS specifically, resulting in a significant reduction of toxicity parameters investigated and a marked increase of the survival preOLs.

摘要

背景

母体-胎儿感染/炎症被认为是脑室周围白质软化症（PVL）发病机制中的一种机制。小胶质细胞（MGs）的激活可能导致前少突胶质细胞损伤。本研究旨在探讨 N-[3-（氨甲基）苄基]乙脒（1400W），一种诱导型一氧化氮合酶（iNOS）的选择性抑制剂，对脂多糖（LPS）诱导的小胶质细胞毒性对前少突胶质细胞（preOLs）的阻断作用。

方法

从两天大的 Sprague-Dawley（SD）新生大鼠中获得共培养的 MGs 和 preOLs，分为三组：共培养对照组、共培养 LPS 组和共培养 LPS 加 1400W 组。通过硝酸还原比色法测量一氧化氮（NO）浓度，免疫细胞化学法测定过氧亚硝酸盐（ONOO（-））水平，Western blot 检测诱导型一氧化氮合酶（iNOS）的合成水平，流式细胞术检测共培养细胞在 LPS（100ng/ml）诱导 48 小时后 preOLs 的凋亡率。

结果

与共培养对照组相比，LPS 诱导的共培养 LPS 组中 NO（82.27+/-3.41μmol/L 对 167.86+/-9.87μmol/L，P<0.01）、ONOO（-）（6.14+/-1.27μmol/L 对 34.38+/-7.75μmol/L，P<0.01）和 iNOS（0.18+/-0.027 对 0.79+/-0.068，P<0.01）水平显著升高，preOLs 的凋亡率较高（6.73+/-1.39%对 24.77+/-2.05%，P<0.01）。在共培养 LPS 加 1400W 组中，用 10μM 1400W 处理后，NO（69.55+/-5.07μmol/L，P<0.01）、ONOO（-）（10.33+/-3.47μmol/L，P<0.01）和 iNOS（0.35+/-0.042，P<0.01）水平显著降低，preOLs 的凋亡率（11.80+/-2.06%对 24.77+/-2.05%，P<0.01）也较共培养 LPS 组降低。