原花青素对脂多糖诱导RAW264.7细胞COX-2酶活性、mRNA及蛋白表达的影响

Effect of proanthocyanidins on COX-2 enzyme activity and COX-2 mRNA /protein expression in LPS-induced RAW264.7 cells

目的 探讨原花青素对脂多糖(LPS)诱导小鼠巨噬细胞株RAW264 7细胞COX 2酶活性及蛋白表达的影响。方法 放射免疫法检测COX 2酶活性,RT PCR检测COX 2mRNA表达,Westernblotting检测COX 2蛋白表达。结果 原花青素( 0 8, 4和20mg·L-1 )不影响LPS诱导RAW264 7细胞COX 2酶活性,可下调LPS诱导RAW264 7细胞COX 2mRNA表达;原花青素( 4和20mg·L-1 )下调LPS诱导RAW264.7细胞COX-2蛋白表达。结论 原花青素不影响LPS诱导RAW264.7细胞COX 2酶活性,但对LPS诱导RAW264 7细胞COX 2mRNA及蛋白表达抑制作用明显。

Aim To study the effect of proanthocyanidins on the COX-2 enzyme activity and COX-2 protein expression in LPS-induced RAW264.7 cells. Methods After being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 (mg·L^(-1)) LPS for 9 h, the effect of proanthocyanidins on the activity of COX-2 enzyme in RAW264.7 cells was analysed by radioimmunoassay(RIA). After being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 (mg·L^(-1)) LPS for 9 h, the effects of proanthocyanidins on the expressions of COX-2 mRNA and protein in RAW264.7 cells were analysed by RT-PCR and Western blotting. Results The activity of COX-2 enzyme was not inhibited by proanthocyanidins (0.8, 4 and 20 (mg·L^(-1)), P>0.05 vs LPS group), but the activity of COX-2 enzyme was significantly inhibited by 10 (μmol·L^(-1)) NS-398 (P<0.01 vs LPS group). The expression of COX-2 mRNA was inhibited by proanthocyanidins (0.8, 4 and 20 (mg·L^(-1))). The expression of COX-2 protein was inhibited by proanthocyanidins (4 and 20 (mg·L^(-1))). Conclusion Proanthocyanidins had no effect on the activity of COX-2 enzyme in LPS-induced RAW264.7 cells. Proanthocyanidins inhibited significantly the expression of COX-2 mRNA and protein in LPS-induced RAW264.7 cells.