Withaferin A Inhibits Nuclear Factor-κB-Dependent Pro-Inflammatory and Stress Response Pathways in the Astrocytes

Figure 4

Pre-treatment with Withaferin A reduces the expression of TNFα, COX-2, and iNOS triggered by LPS in primary spinal cord astrocytes. Primary astrocytes were pre-treated with 10 μM SC-514, a selective NF-κB inhibitor, or 0.5 μM Withaferin A (WA) for 1 hour and then incubated in the presence or in the absence of LPS (1 μg/mL) for 6 hours. Total RNA was extracted and reverse-transcribed. TNFα, COX-2, and iNOS mRNA levels were analyzed by RT-qPCR ( experiments in triplicate). Values (mean ± s.e.m.) were normalized relative to HPRT and expressed as percentage of TNFα, COX-2, or iNOS expression levels in control conditions ( versus control, and versus LPS; one-way ANOVA followed by Bonferroni post hoc test).