An ethyl acetate fraction of Moringa oleifera Lam. Inhibits human macrophage cytokine production induced by cigarette smoke - PubMed (original) (raw)

An ethyl acetate fraction of Moringa oleifera Lam. Inhibits human macrophage cytokine production induced by cigarette smoke

Nateelak Kooltheat et al. Nutrients. 2014.

Abstract

Moringa oleifera Lam. (MO) has been reported to harbor anti-oxidation and anti-inflammatory activity and useful in the treatment of inflammatory diseases. However, despite these findings there has been little work done on the effects of MO on immune cellular function. Since macrophages, TNF and related cytokines play an important pathophysiologic role in lung damage induced by cigarette smoke, we examined the effects of MO on cigarette smoke extract (CSE)-induced cytokine production by human macrophages. An ethyl acetate fraction of MO (MOEF) was prepared from fresh leaves extract of Moringa and shown to consist of high levels of phenolic and antioxidant activities. Human monocyte derived macrophages (MDM) pre-treated with varying concentrations of MOEF showed decreased production of TNF, IL-6 and IL-8 in response to both LPS and CSE. The decrease was evident at both cytokine protein and mRNA levels. Furthermore, the extract inhibited the expression of RelA, a gene implicated in the NF-κB p65 signaling in inflammation. The findings highlight the ability of MOEF to inhibit cytokines (IL-8) which promote the infiltration of neutrophils into the lungs and others (TNF, IL-6) which mediate tissue disease and damage.

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Figures

Figure 1

Figure 1

Fractionation of the phenolic and antioxidant activity of Moringa oleifera. Total phenolic content was measured by Folin-Ciocalteau method and expressed as mg Pyrogallol equivalen/g of dry extract (□). Antioxidant was measured by ABTS radical cation decolorization assay and expressed as μM Trolox equivalent/100 mg of dry extract (■). Results were analyzed from triplicate data of experiments. * p < 0.01 by ANOVA, compared to crude extract and other fractions.

Figure 2

Figure 2

Dose-response curves of each test substances, showing LC50 and LC10 of NIC; nicotine (A); CSE; cigarette smoke extract (B); ASA; Aspirin (C); and MOEF; ethyl acetate fraction from Moringa oleifera extract (D). Cells were treated with 2-fold pre-diluted substances for 12 h and tested for their cellular cytotoxicity by neutral red uptake cytotoxicity bioassay. Data of cell viability in triplicate were used to calculate LC50 and LC10 of each substance by dose-response/sigmoidal curve fitting analysis.

Figure 3

Figure 3

Effects of MOEF on LPS- and CSE-induced tumor necrotic factor alpha gene (TNF), IL-6 and IL-8 production by human MDM. Cells were pre-treated with MOEF, ASA or diluent for 6 h and then stimulated with LPS or CSE. After further 12 h incubation, culture supernatants were harvested to quantify the cytokines. Results are presented as Mean ± SEM of three experiments each conducted with cell from different individual and each in triplicate. Statistical analyses; * p < 0.01 by ANOVA, compared to non stimulated control.

Figure 4

Figure 4

Effects of MOEF on LPS- and CSE-induced TNF, IL-6, IL-8 and RelA gene expression in human MDM. Cells were pre-treated with MOEF, ASA or diluent for 6 h and then stimulated with LPS or CSE. After a further 12 h incubation culture supernatants were harvested to quantify the cytokines mRNA by one-step qRT-PCR. Results are presented as Mean ± SEM of normalized data of three experiments each conducted with cell from different individual and each in triplicate. Statistical analyses; * p < 0.01 by ANOVA, compared to non stimulated control.

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