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Effect of 1400W on blocking lipopolysaccharide-induced microglial toxicity to preoligodendrocytes
Ya-Fang He1, Hui-Jin Chen, Long-Huan Qian
1Shanghai Institute for Pediatric Research, Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.
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.
Insights
N-[3-(aminomethyl) benzyl] acetamidine (1400W) effectively reduces lipopolysaccharide-induced microglial toxicity to preoligodendrocytes by inhibiting inducible nitric oxide synthase (iNOS). This inhibition significantly lowers toxicity markers and increases preoligodendrocyte survival, offering a potential therapeutic strategy for conditions like periventricular leukomalacia.
Area of Science:
- Neuroscience
- Developmental Biology
- Immunology
Background:
- Maternal-fetal infection/inflammation is a suspected mechanism in periventricular leukomalacia (PVL) pathogenesis.
- Microglial (MG) activation may lead to preoligodendroglial (preOL) damage.
- N-[3-(aminomethyl) benzyl] acetamidine (1400W), a selective inducible nitric oxide synthase (iNOS) inhibitor, was investigated for its effect on lipopolysaccharide (LPS)-induced microglial toxicity to preOLs.
Purpose of the Study:
- To explore the efficacy of 1400W in blocking LPS-induced microglial toxicity to preOLs.
- To investigate the role of iNOS inhibition in mitigating neuroinflammation-related preOL damage.
Main Methods:
- Co-culture of Sprague-Dawley neonatal rat microglias (MGs) and preoligodendrocytes (preOLs).
- Groups included: co-culture control, co-culture LPS-induced, and co-culture LPS + 1400W.
- Measurements: nitric oxide (NO), peroxynitrite (ONOO-), iNOS levels, and preOL apoptosis rate via flow cytometry.
Main Results:
- LPS significantly increased NO, ONOO-, iNOS levels, and preOL apoptosis compared to controls (P<0.01).
- Treatment with 10 micromol/L 1400W significantly reduced NO, ONOO-, and iNOS levels (P<0.01).
- 1400W treatment also significantly decreased preOL apoptosis rate compared to the LPS-only group (P<0.01).
Conclusions:
- 1400W effectively blocks LPS-induced microglial toxicity to preOLs by specifically inhibiting iNOS.
- This inhibition leads to reduced toxicity markers and increased preOL survival.
- The findings suggest 1400W as a potential therapeutic agent for PVL and related conditions.
