ginseng.
To further investigate the active immunomodulatory compounds of ginseng, the aqueous and polysaccharide extracts were fractioned using differential fractionation with ultra centrifugal filter units. The aqueous and polysaccharide extracts were chosen as they induced a much greater response in cytokine production, as well as were the only extracts to have an effect on the SEE response. When PBMC were treated with the fractions of the aqueous and polysaccharide extracts, the only fraction which was able to come close to recapitulating the IL-6 response of the extracts themselves was the greater than 100 kDa fraction. Furthermore, the other four fractions did not induce a similar profile of cytokine production; the production induced by these fractions was much less than that of the extracts themselves. This result was reflected in both the aqueous and polysaccharide extracts. Moreover, the ability of the fractions to induce the production of other cytokines including IL-1β and TNF-α was measured also, with similar trends as the IL-6 data presented.
In order to further understand the effect of the greater than 100 kDa fraction in its capacity to immunomodulate, we determined the effect of the fractions compared to the aqueous and polysaccharide ginseng extracts on the SEE response. PBMC were co- treated with SEE and the different ginseng fractions from the aqueous and polysaccharide extracts, as well as the extracts themselves. Similar to the previously observed profile, only the greater then 100 kDa fraction had a similar effect on the SEE response as the extracts.
Ginsenosides, low molecular weight saponin molecules unique to GS, have historically been considered the active immunomodulatory ingredients of ginseng. However, since
this conflicted with our previous results, we obtained 11 purified, biologically active ginsenosides and two ginsenoside mixtures to test their immunomodulatory capacity. C- Y, C-Mc and Gyp LXXV (pure) induced the pro-inflammatory cytokine IL-1β, while C- Y, C-Mc, Gyp LXXV (pure), C-O and Gyp XVII (mixture) induced significant IL-6 production in PBMC. However, contrary to the effects of the ginseng extracts, all of the 13 ginsenosides tested decreased IL-6 production in response to LPS. Alternatively, none of the ginsenosides tested had a biologically relevant effect on the response to SEE. The observation that ginsenosides down-regulate the LPS response while the aqueous and polysaccharide extracts had no effect on the LPS response, implied the existence of a possible antagonistic effect between ginsenosides and the other components of the extracts, in particular the polysaccharides in the ginseng extracts. To determine if this was the case, PBMC were co-stimulated with ginsenosides and the aqueous or
polysaccharide extract. While none of the ginsenosides tested antagonized IL-1β induced by the polysaccharide ginseng extract, C-Mc1, C-K, C-Mc, C-O, C-Y, Gyp XVII (pure), F2, PPD, Rg3 and Rh2 antagonized the IL-1β induced by the aqueous extract.
The antagonism of the aqueous ginseng extract prompted us to further characterize the effects of ginsenosides on PBMC treated with either the aqueous or polysaccharide extract when challenged with LPS or SEE. All of the 13 ginsenosides tested except Gyp LXXV (mixture), Gyp XVII (mixture) and Gyp LXXV (pure) antagonized the IL-6 production of PBMC treated with LPS, the aqueous ginseng extract and a ginsenoside, when compared to PBMC treated with LPS and the aqueous ginseng extract.
Furthermore, there was no meaningful biological difference between PBMC treated with LPS, the polysaccharide ginseng extract and a ginsenoside when compared to PBMC treated with LPS and the polysaccharide ginseng extract. Similarly, when the IL-2
production of PBMC treated with SEE, the aqueous ginseng extract and ginsenosides was compared to that of PBMC treated with SEE and the aqueous ginseng extract, Gyp XVII (mixture), F2, Rh2 were the only ginsenosides that did not result in an increase in IL-2 production. While, when PBMC were treated with SEE, the polysaccharide ginseng extract and a ginsenoside was compared with the IL-2 production of PBMC treated with SEE and the polysaccharide ginseng extract, only Gyp XVII (mixture) and Gyp XVII
(pure) resulted in an increase in IL-2 production. Based on our results it is likely that the antagonism of ginseng polysaccharides by ginsenosides occurs at a common step
downstream of the binding receptor for each compound. This also has major implications for the use of ginseng extracts and it may be more beneficial to use single compounds of ginseng based on the desired outcomes. For example, ginseng polysaccharides may be more beneficial for increasing pro-inflammatory cytokine secretion, while ginsenosides decrease the immune response to LPS.
1.7
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