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Genomics and proteomics of immune modulatory effects of a butanol fraction of echinacea purpurea in human dendritic cells.

Wang CY, Staniforth V, Chiao MT, Hou CC, Wu HM, Yeh KC, Chen CH, Hwang PI, Wen TN, Shyur LF, Yang NS - BMC Genomics (2008)

Bottom Line: Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8.Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

View Article: PubMed Central - HTML - PubMed

Affiliation: Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan. sally@mail.biowell.com.tw

ABSTRACT

Background: Echinacea spp. extracts and the derived phytocompounds have been shown to induce specific immune cell activities and are popularly used as food supplements or nutraceuticals for immuno-modulatory functions. Dendritic cells (DCs), the most potent antigen presenting cells, play an important role in both innate and adaptive immunities. In this study, we investigated the specific and differential gene expression in human immature DCs (iDCs) in response to treatment with a butanol fraction containing defined bioactive phytocompounds extracted from stems and leaves of Echinacea purpurea, that we denoted [BF/S+L/Ep].

Results: Affymetrix DNA microarray results showed significant up regulation of specific genes for cytokines (IL-8, IL-1beta, and IL-18) and chemokines (CXCL 2, CCL 5, and CCL 2) within 4 h after [BF/S+L/Ep] treatment of iDCs. Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8. Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.

Conclusion: This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

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Confirmation of the 2-D gel proteomic results on down-regulation of cofilin protein expression with Western blot analysis. (A) Differential expression of cofilin in human iDCs treated with [BF/S+L/Ep] (75 μg/ml), cichoric acid (50 μg/ml) or vehicle control, at 12 and 24 h treatment, as revealed by 2-D gel electrophoresis. (B) Western blot analysis of expression of cofilin in human iDCs. Other experimental details are the same as described in Figure 7. Similar trends were observed in three independent experiments.
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Figure 8: Confirmation of the 2-D gel proteomic results on down-regulation of cofilin protein expression with Western blot analysis. (A) Differential expression of cofilin in human iDCs treated with [BF/S+L/Ep] (75 μg/ml), cichoric acid (50 μg/ml) or vehicle control, at 12 and 24 h treatment, as revealed by 2-D gel electrophoresis. (B) Western blot analysis of expression of cofilin in human iDCs. Other experimental details are the same as described in Figure 7. Similar trends were observed in three independent experiments.

Mentions: Western blot analysis was conducted to confirm some of the up-regulated proteins observed from proteomic analysis. The expression of Mn-SOD (SOD2) was increased 2.65- to 1.99-fold with [BF/S+L/Ep] and 2.35- and 1.67-fold with cichoric acid treatment at 12 and 24 h, respectively, as compared with vehicle controls, which is consistent with results from 2-D gel electrophoresis (Figure 7, Table 4). The levels of cofilin determined by 2-D gel electrophoresis (Figure 8A) were slightly decreased, by 0.75- and 0.84-fold, with [BF/S+L/Ep] and greatly decreased, by 0.09- and 0.42-fold, with cichoric acid treatment at 12 or 24 h, respectively. Interestingly, confirmation by Western blot analysis revealed only a 0.5- and 0.32-fold decrease in cofilin level at 12 and 24 h with cichoric acid treatment (Figure 8B). Both assay systems revealed reduced effect of [BF/S+L/Ep] on cofilin expression in iDCs (0.75- and 0.84-fold, respectively) (Figure 8A Vs 8B). Nevertheless, these results confirmed our finding that the expression of cofilin in iDCs can be substantially reduced by treatment with cichoric acid and may be slightly inhibited by [BF/S+L/Ep]. Rutin, another major index component of the [BF/S+L/Ep], effectively inhibited the expression of cofilin, by 0.4- and 0.6-fold at 12 and 24 h, respectively, in iDCs (Figure 8B). Treatment with 100 ng/ml lipopolysaccharide (LPS, used as a positive control) conferred an approximately 0.5- to 0.4-fold decrease in cofilin expression as compared to controls (Figure 8B).


Genomics and proteomics of immune modulatory effects of a butanol fraction of echinacea purpurea in human dendritic cells.

Wang CY, Staniforth V, Chiao MT, Hou CC, Wu HM, Yeh KC, Chen CH, Hwang PI, Wen TN, Shyur LF, Yang NS - BMC Genomics (2008)

Confirmation of the 2-D gel proteomic results on down-regulation of cofilin protein expression with Western blot analysis. (A) Differential expression of cofilin in human iDCs treated with [BF/S+L/Ep] (75 μg/ml), cichoric acid (50 μg/ml) or vehicle control, at 12 and 24 h treatment, as revealed by 2-D gel electrophoresis. (B) Western blot analysis of expression of cofilin in human iDCs. Other experimental details are the same as described in Figure 7. Similar trends were observed in three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2571112&req=5

Figure 8: Confirmation of the 2-D gel proteomic results on down-regulation of cofilin protein expression with Western blot analysis. (A) Differential expression of cofilin in human iDCs treated with [BF/S+L/Ep] (75 μg/ml), cichoric acid (50 μg/ml) or vehicle control, at 12 and 24 h treatment, as revealed by 2-D gel electrophoresis. (B) Western blot analysis of expression of cofilin in human iDCs. Other experimental details are the same as described in Figure 7. Similar trends were observed in three independent experiments.
Mentions: Western blot analysis was conducted to confirm some of the up-regulated proteins observed from proteomic analysis. The expression of Mn-SOD (SOD2) was increased 2.65- to 1.99-fold with [BF/S+L/Ep] and 2.35- and 1.67-fold with cichoric acid treatment at 12 and 24 h, respectively, as compared with vehicle controls, which is consistent with results from 2-D gel electrophoresis (Figure 7, Table 4). The levels of cofilin determined by 2-D gel electrophoresis (Figure 8A) were slightly decreased, by 0.75- and 0.84-fold, with [BF/S+L/Ep] and greatly decreased, by 0.09- and 0.42-fold, with cichoric acid treatment at 12 or 24 h, respectively. Interestingly, confirmation by Western blot analysis revealed only a 0.5- and 0.32-fold decrease in cofilin level at 12 and 24 h with cichoric acid treatment (Figure 8B). Both assay systems revealed reduced effect of [BF/S+L/Ep] on cofilin expression in iDCs (0.75- and 0.84-fold, respectively) (Figure 8A Vs 8B). Nevertheless, these results confirmed our finding that the expression of cofilin in iDCs can be substantially reduced by treatment with cichoric acid and may be slightly inhibited by [BF/S+L/Ep]. Rutin, another major index component of the [BF/S+L/Ep], effectively inhibited the expression of cofilin, by 0.4- and 0.6-fold at 12 and 24 h, respectively, in iDCs (Figure 8B). Treatment with 100 ng/ml lipopolysaccharide (LPS, used as a positive control) conferred an approximately 0.5- to 0.4-fold decrease in cofilin expression as compared to controls (Figure 8B).

Bottom Line: Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8.Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

View Article: PubMed Central - HTML - PubMed

Affiliation: Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan. sally@mail.biowell.com.tw

ABSTRACT

Background: Echinacea spp. extracts and the derived phytocompounds have been shown to induce specific immune cell activities and are popularly used as food supplements or nutraceuticals for immuno-modulatory functions. Dendritic cells (DCs), the most potent antigen presenting cells, play an important role in both innate and adaptive immunities. In this study, we investigated the specific and differential gene expression in human immature DCs (iDCs) in response to treatment with a butanol fraction containing defined bioactive phytocompounds extracted from stems and leaves of Echinacea purpurea, that we denoted [BF/S+L/Ep].

Results: Affymetrix DNA microarray results showed significant up regulation of specific genes for cytokines (IL-8, IL-1beta, and IL-18) and chemokines (CXCL 2, CCL 5, and CCL 2) within 4 h after [BF/S+L/Ep] treatment of iDCs. Bioinformatics analysis of genes expressed in [BF/S+L/Ep]-treated DCs revealed a key-signaling network involving a number of immune-modulatory molecules leading to the activation of a downstream molecule, adenylate cyclase 8. Proteomic analysis showed increased expression of antioxidant and cytoskeletal proteins after treatment with [BF/S+L/Ep] and cichoric acid.

Conclusion: This study provides information on candidate target molecules and molecular signaling mechanisms for future systematic research into the immune-modulatory activities of an important traditional medicinal herb and its derived phytocompounds.

Show MeSH
Related in: MedlinePlus