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Evaluation of in vitro anti-inflammatory effects of crude ginger and rosemary extracts obtained through supercritical CO2 extraction on macrophage and tumor cell line: the influence of vehicle type.

Justo OR, Simioni PU, Gabriel DL, Tamashiro WM, Rosa Pde T, Moraes ÂM - BMC Complement Altern Med (2015)

Bottom Line: Controversially, rosemary extract dispersed on DMSO induced a more significant IL-1 and TNF-α reduction than ginger extract in primary macrophages.It can be concluded that some of the beneficial effects attributed to extracts of ginger and rosemary may be associated with the inhibition of inflammatory mediators due to their high antioxidant activity.However, these effects were influenced by the type of delivery vehicle.

View Article: PubMed Central - PubMed

Affiliation: Department of Engineering of Materials and of Bioprocesses - School of Chemical Engineering, University of Campinas, 13083-852, Campinas, SP, Brazil.

ABSTRACT

Background: Numerous plants from have been investigated due to their anti-inflammatory activity and, among then, extracts or components of ginger (Zingiber officinale Roscoe) and rosemary (Rosmarinus officinalis L.), sources of polyphenolic compounds. 6-gingerol from ginger rhizome and carnosic acid and carnosol from rosemary leaves present anti-tumor, anti-inflammatory and antioxidant activities. However, the evaluation of the mechanisms of action of these and other plant extracts is limited due to their high hydrophobicity. Dimethylsulfoxide (DMSO) is commonly used as a vehicle of liposoluble materials to mammalian cells in vitro, presenting enhanced cell penetration. Liposomes are also able to efficiently deliver agents to mammalian cells, being capable to incorporate in their structure not only hydrophobic molecules, but also hydrophilic and amphiphilic compounds. Another strategy is based on the use of Pluronic F-68, a biocompatible low-foaming, non-ionic surfactant, to disperse hydrophobic components. Here, these three delivery approaches were compared to analyze their influence on the in vitro anti-inflammatory effects of ginger and rosemary extracts, at different concentrations, on primary mammalian cells and on a tumor cell line.

Methods: Ginger and rosemary extracts free of organic solvents were obtained by supercritical fluid extraction and dispersed in DMSO, Pluronic F-68 or liposomes, in variable concentrations. Cell viability, production of inflammatory mediators and nitric oxide (NO) release were measured in vitro on J774 cell line and murine macrophages primary culture stimulated with bacterial lipopolysaccharide and interferon-γ after being exposed or not to these extracts.

Results: Ginger and rosemary extracts obtained by supercritical CO2 extraction inhibited the production of pro-inflammatory cytokines and the release of NO by peritoneal macrophages and J774 cells. The delivery vehicles influenced the anti-inflammatory effects. Comparatively, the ginger extract showed the highest anti-inflammatory activity on the tumor cell line. Controversially, rosemary extract dispersed on DMSO induced a more significant IL-1 and TNF-α reduction than ginger extract in primary macrophages.

Conclusions: Amongst the tested delivery vehicles, DMSO was the most suitable, presenting reduced cytotoxicity, followed by Pluronic F-68 and liposomes, provably due to differences in their form of absorption, distribution and cellular metabolism. Co-administration of liposomes and plant extracts may cause death of macrophages cells and induction of NO production. It can be concluded that some of the beneficial effects attributed to extracts of ginger and rosemary may be associated with the inhibition of inflammatory mediators due to their high antioxidant activity. However, these effects were influenced by the type of delivery vehicle.

No MeSH data available.


Related in: MedlinePlus

Comparison of the effects of ginger and rosemary extracts dissolved in DMSO on nitric oxide release and inflammatory cytokine production by stimulated murine peritoneal and J774 macrophages cells. Nitric oxide production (a) and levels of TNF-α (b) and IL-1 (c) cytokines secreted by murine peritoneal or J774 macrophages cells incubated for 48 h with rosemary and ginger extracts dissolved in DMSO and stimulated or not with LPS/IFN-γ: LPS (1 μg/mL) plus IFN- γ (150 IU/mL). Cells were treated with extracts at 0.7 mg/mL. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM (n = 6). * indicates data statistically significantly different in comparison with the control (no-treated cells) at p < 0.05
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Fig5: Comparison of the effects of ginger and rosemary extracts dissolved in DMSO on nitric oxide release and inflammatory cytokine production by stimulated murine peritoneal and J774 macrophages cells. Nitric oxide production (a) and levels of TNF-α (b) and IL-1 (c) cytokines secreted by murine peritoneal or J774 macrophages cells incubated for 48 h with rosemary and ginger extracts dissolved in DMSO and stimulated or not with LPS/IFN-γ: LPS (1 μg/mL) plus IFN- γ (150 IU/mL). Cells were treated with extracts at 0.7 mg/mL. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM (n = 6). * indicates data statistically significantly different in comparison with the control (no-treated cells) at p < 0.05

Mentions: Pro-inflammatory cytokines stimulate the generation of reactive oxygen and nitrogen species [27]. Therefore, the production of such reactive species in macrophages is likely to be inhibited when the generation of pro-inflammatory cytokines is reduced. Since the tested ginger and rosemary extracts dispersed in DMSO induced the highest inhibition on NO production in specific conditions, the effect of these extracts in DMSO on pro-inflammatory cytokine production by peritoneal macrophages and J774 cells was evaluated (Fig. 5). For the IL-1 and TNF-α assays, the plant extracts were used only at concentrations in which cell proliferation (evaluated by the MTT assay) was not affected (equal to 0.7 mg/mL) to exclude the possibility that the observed effects of the extracts were due to cytotoxicity.Fig. 5


Evaluation of in vitro anti-inflammatory effects of crude ginger and rosemary extracts obtained through supercritical CO2 extraction on macrophage and tumor cell line: the influence of vehicle type.

Justo OR, Simioni PU, Gabriel DL, Tamashiro WM, Rosa Pde T, Moraes ÂM - BMC Complement Altern Med (2015)

Comparison of the effects of ginger and rosemary extracts dissolved in DMSO on nitric oxide release and inflammatory cytokine production by stimulated murine peritoneal and J774 macrophages cells. Nitric oxide production (a) and levels of TNF-α (b) and IL-1 (c) cytokines secreted by murine peritoneal or J774 macrophages cells incubated for 48 h with rosemary and ginger extracts dissolved in DMSO and stimulated or not with LPS/IFN-γ: LPS (1 μg/mL) plus IFN- γ (150 IU/mL). Cells were treated with extracts at 0.7 mg/mL. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM (n = 6). * indicates data statistically significantly different in comparison with the control (no-treated cells) at p < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4625945&req=5

Fig5: Comparison of the effects of ginger and rosemary extracts dissolved in DMSO on nitric oxide release and inflammatory cytokine production by stimulated murine peritoneal and J774 macrophages cells. Nitric oxide production (a) and levels of TNF-α (b) and IL-1 (c) cytokines secreted by murine peritoneal or J774 macrophages cells incubated for 48 h with rosemary and ginger extracts dissolved in DMSO and stimulated or not with LPS/IFN-γ: LPS (1 μg/mL) plus IFN- γ (150 IU/mL). Cells were treated with extracts at 0.7 mg/mL. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM (n = 6). * indicates data statistically significantly different in comparison with the control (no-treated cells) at p < 0.05
Mentions: Pro-inflammatory cytokines stimulate the generation of reactive oxygen and nitrogen species [27]. Therefore, the production of such reactive species in macrophages is likely to be inhibited when the generation of pro-inflammatory cytokines is reduced. Since the tested ginger and rosemary extracts dispersed in DMSO induced the highest inhibition on NO production in specific conditions, the effect of these extracts in DMSO on pro-inflammatory cytokine production by peritoneal macrophages and J774 cells was evaluated (Fig. 5). For the IL-1 and TNF-α assays, the plant extracts were used only at concentrations in which cell proliferation (evaluated by the MTT assay) was not affected (equal to 0.7 mg/mL) to exclude the possibility that the observed effects of the extracts were due to cytotoxicity.Fig. 5

Bottom Line: Controversially, rosemary extract dispersed on DMSO induced a more significant IL-1 and TNF-α reduction than ginger extract in primary macrophages.It can be concluded that some of the beneficial effects attributed to extracts of ginger and rosemary may be associated with the inhibition of inflammatory mediators due to their high antioxidant activity.However, these effects were influenced by the type of delivery vehicle.

View Article: PubMed Central - PubMed

Affiliation: Department of Engineering of Materials and of Bioprocesses - School of Chemical Engineering, University of Campinas, 13083-852, Campinas, SP, Brazil.

ABSTRACT

Background: Numerous plants from have been investigated due to their anti-inflammatory activity and, among then, extracts or components of ginger (Zingiber officinale Roscoe) and rosemary (Rosmarinus officinalis L.), sources of polyphenolic compounds. 6-gingerol from ginger rhizome and carnosic acid and carnosol from rosemary leaves present anti-tumor, anti-inflammatory and antioxidant activities. However, the evaluation of the mechanisms of action of these and other plant extracts is limited due to their high hydrophobicity. Dimethylsulfoxide (DMSO) is commonly used as a vehicle of liposoluble materials to mammalian cells in vitro, presenting enhanced cell penetration. Liposomes are also able to efficiently deliver agents to mammalian cells, being capable to incorporate in their structure not only hydrophobic molecules, but also hydrophilic and amphiphilic compounds. Another strategy is based on the use of Pluronic F-68, a biocompatible low-foaming, non-ionic surfactant, to disperse hydrophobic components. Here, these three delivery approaches were compared to analyze their influence on the in vitro anti-inflammatory effects of ginger and rosemary extracts, at different concentrations, on primary mammalian cells and on a tumor cell line.

Methods: Ginger and rosemary extracts free of organic solvents were obtained by supercritical fluid extraction and dispersed in DMSO, Pluronic F-68 or liposomes, in variable concentrations. Cell viability, production of inflammatory mediators and nitric oxide (NO) release were measured in vitro on J774 cell line and murine macrophages primary culture stimulated with bacterial lipopolysaccharide and interferon-γ after being exposed or not to these extracts.

Results: Ginger and rosemary extracts obtained by supercritical CO2 extraction inhibited the production of pro-inflammatory cytokines and the release of NO by peritoneal macrophages and J774 cells. The delivery vehicles influenced the anti-inflammatory effects. Comparatively, the ginger extract showed the highest anti-inflammatory activity on the tumor cell line. Controversially, rosemary extract dispersed on DMSO induced a more significant IL-1 and TNF-α reduction than ginger extract in primary macrophages.

Conclusions: Amongst the tested delivery vehicles, DMSO was the most suitable, presenting reduced cytotoxicity, followed by Pluronic F-68 and liposomes, provably due to differences in their form of absorption, distribution and cellular metabolism. Co-administration of liposomes and plant extracts may cause death of macrophages cells and induction of NO production. It can be concluded that some of the beneficial effects attributed to extracts of ginger and rosemary may be associated with the inhibition of inflammatory mediators due to their high antioxidant activity. However, these effects were influenced by the type of delivery vehicle.

No MeSH data available.


Related in: MedlinePlus