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Statistical analysis. The results were expressed as means ± SEM from at least three independent experiments. Comparisons between groups were performed using Student’s t-test, and analysis of variance (ANOVA) was used for comparisons among groups. A p-value of less than 0.05 was considered to be statistically significant.
Results
Screening, isolation, and identification of PCG and its effects on NFAT activation
From the natural product library screening, a herbal extract from the dried fruits of P. granatum (pomegranate) showed more than 90% inhibition in the NFAT reporter assay at 100 μg/ml, and it was non-toxic to NFAT-Jurkat (-)-MK 801 and human PBMC at the same concentration (results not shown). The active compound was identified as PCG based on the spectral data from the 1H and 13C NMR, and its structure was confirmed by comparison with the former literature data ( Fig. 1A) [8]. The immune-suppressive activity of isolated PCG was examined in the NFAT reporter assay. PCG reduced the SEAP activity significantly in a dose-dependent manner (p