Limits...
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

Effects of ginger and rosemary extracts on the viability and nitric oxide production by LPS and/or IFN-γ-stimulated J774 macrophages cells: influence of vehicle and concentration of extracts. Cells were incubated for 48 h with ginger and rosemary extracts dissolved in DMSO (a) and Pluronic-F68 (b) and stimulated with LPS/IFN-γ: LPS (1 μg/mL) plus IFN-γ (150 IU/mL). Cells were treated with extracts at the indicated concentrations. Cell viability was determined by MTT assay. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM of the data from triplicate tests. * 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
getmorefigures.php?uid=PMC4625945&req=5

Fig4: Effects of ginger and rosemary extracts on the viability and nitric oxide production by LPS and/or IFN-γ-stimulated J774 macrophages cells: influence of vehicle and concentration of extracts. Cells were incubated for 48 h with ginger and rosemary extracts dissolved in DMSO (a) and Pluronic-F68 (b) and stimulated with LPS/IFN-γ: LPS (1 μg/mL) plus IFN-γ (150 IU/mL). Cells were treated with extracts at the indicated concentrations. Cell viability was determined by MTT assay. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM of the data from triplicate tests. * indicates data statistically significantly different in comparison with the control (no-treated cells) at p < 0.05

Mentions: As depicted in Fig. 4a, treatment of J774 cells with LPS + IFN-γ did not result in significant proliferation decline when DMSO was used as a vehicle. Treatment of J774 cells with ginger and rosemary extracts dissolved in DMSO inhibited LPS-induced NO production in a dose-dependent manner at concentration range from 0.04 to 2.79 mg/mL. Treatment of J774 cells with LPS + IFN-γ and ginger extract dissolved in Pluronic-F68 caused a significant proliferation decline in a dose-dependent manner at concentration range from 0.17 to 2.79 mg/mL, whereas significant cell death was noted only when J774 cells were exposed to the highest concentration of rosemary (2.79 mg/mL) extract (Fig. 4b). The exposure of J774 cells to LPS plus IFN-γ resulted in the production of high amounts of NO, which was inhibited only by rosemary extract dispersed in Pluronic F-68.Fig. 4


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)

Effects of ginger and rosemary extracts on the viability and nitric oxide production by LPS and/or IFN-γ-stimulated J774 macrophages cells: influence of vehicle and concentration of extracts. Cells were incubated for 48 h with ginger and rosemary extracts dissolved in DMSO (a) and Pluronic-F68 (b) and stimulated with LPS/IFN-γ: LPS (1 μg/mL) plus IFN-γ (150 IU/mL). Cells were treated with extracts at the indicated concentrations. Cell viability was determined by MTT assay. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM of the data from triplicate tests. * 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

Fig4: Effects of ginger and rosemary extracts on the viability and nitric oxide production by LPS and/or IFN-γ-stimulated J774 macrophages cells: influence of vehicle and concentration of extracts. Cells were incubated for 48 h with ginger and rosemary extracts dissolved in DMSO (a) and Pluronic-F68 (b) and stimulated with LPS/IFN-γ: LPS (1 μg/mL) plus IFN-γ (150 IU/mL). Cells were treated with extracts at the indicated concentrations. Cell viability was determined by MTT assay. The amount of NO released into the culture supernatants is expressed as nitrite. The columns represent the means ± SEM of the data from triplicate tests. * indicates data statistically significantly different in comparison with the control (no-treated cells) at p < 0.05
Mentions: As depicted in Fig. 4a, treatment of J774 cells with LPS + IFN-γ did not result in significant proliferation decline when DMSO was used as a vehicle. Treatment of J774 cells with ginger and rosemary extracts dissolved in DMSO inhibited LPS-induced NO production in a dose-dependent manner at concentration range from 0.04 to 2.79 mg/mL. Treatment of J774 cells with LPS + IFN-γ and ginger extract dissolved in Pluronic-F68 caused a significant proliferation decline in a dose-dependent manner at concentration range from 0.17 to 2.79 mg/mL, whereas significant cell death was noted only when J774 cells were exposed to the highest concentration of rosemary (2.79 mg/mL) extract (Fig. 4b). The exposure of J774 cells to LPS plus IFN-γ resulted in the production of high amounts of NO, which was inhibited only by rosemary extract dispersed in Pluronic F-68.Fig. 4

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