Gelam Honey Inhibits the Production of Proinflammatory, Mediators NO, PGE(2), TNF-α, and IL-6 in Carrageenan-Induced Acute Paw Edema in Rats

Saba Zuhair Hussein, Kamaruddin Mohd Yusoff, Suzana Makpol, Yasmin Anum Mohd Yusof, Saba Zuhair Hussein, Kamaruddin Mohd Yusoff, Suzana Makpol, Yasmin Anum Mohd Yusof

Abstract

Natural honey is well known for its therapeutic value and has been used in traditional medicine of different cultures throughout the world. The aim of this study was to investigate the anti-inflammatory effect of Malaysian Gelam honey in inflammation-induced rats. Paw edema was induced by a subplantar injection of 1% carrageenan into the rat right hind paw. Rats were treated with the nonsteroidal anti-inflammatory drug (NSAID) Indomethacin (10 mg/kg, p.o.) or Gelam honey at different doses (1 or 2 g/kg, p.o.). The increase in footpad thickness was considered to be edema, which was measured using a dial caliper. Plasma and paw tissue were collected to analyze the production of inflammatory mediators, such as NO, PGE(2), TNF-α, and IL-6, as well as iNOS and COX-2. The results showed that Gelam honey could reduce edema in a dose-dependent fashion in inflamed rat paws, decrease the production of NO, PGE(2), TNF-α, and IL-6 in plasma, and suppress the expression of iNOS, COX-2, TNF-α, and IL-6 in paw tissue. Oral pretreatment of Gelam honey at 2 g/kg of body weight at two time points (1 and 7 days) showed a significantly decreased production of proinflammatory cytokines, which was similar to the effect of the anti-inflammatory drug Indomethacin (NSAID), both in plasma and tissue. Thus, our results suggest that Gelam honey has anti-inflammatory effects by reducing the rat paw edema size and inhibiting the production of proinflammatory mediators. Gelam honey is potentially useful for treating inflammatory conditions.

Figures

Figure 1
Figure 1
Representative photographs of rats' right hind paw (a) before (b) after 5 hr and (c) after 24 hr of carrageenan-induced paw edema inflammation.
Figure 2
Figure 2
The effect of Gelam honey on the paw size in carrageenan-induced paw edema in rats. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for (a) 1 day or (b) 7 days before carrageenan injection. D.W.: distilled water, Carr: Carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). *Significantly (P < 0.05) different compared to the control (D.W. + Carr).
Figure 3
Figure 3
The effect of Gelam honey on NO production. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation, as induced by carrageenan injection. D.W.: distilled water, Carr: Carrageenan, IND: Indomethacin. Data were presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared at different time points, 1 day and 7 days.
Figure 4
Figure 4
The effect of Gelam honey on PGE2 production. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation, as induced by carrageenan injection. D.W.: distilled water, Carr: Carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared between different time points, 1 day and 7 days.
Figure 5
Figure 5
The effect of Gelam honey on TNF-α production. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation, as induced by carrageenan injection. D.W.: distilled water, Carr: Carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared at different time points, 1 day and 7 days.
Figure 6
Figure 6
The effect of Gelam honey on IL-6 production. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation, as induced by carrageenan injection. D.W.: distilled water, Carr: carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared between different time points, 1 day and 7 days.
Figure 7
Figure 7
The effect of Gelam honey on the gene expression of (a) iNOS, (b) COX-2, (c) TNF-α, and (d) IL-6 in paw tissue. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation, as induced by carrageenan injection. D.W.: distilled water, Carr: carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared between different time points, 1 day and 7 days.
Figure 8
Figure 8
The effect of Gelam honey on the expression of iNOS protein in paw tissue. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation was induced by carrageenan injection. D.W.: distilled water, Carr: carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared at different time points, 1 day and 7 days.
Figure 9
Figure 9
The effect of Gelam honey on the expression of the COX-2 protein in paw tissue. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation was induced by carrageenan injection. D.W.: distilled water, Carr: carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared at different time points, 1 day and 7 days.
Figure 10
Figure 10
The effect of Gelam honey on the expression of TNF-α protein in paw tissue. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation was induced by carrageenan injection. D.W.: distilled water, Carr: carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared at different time points, 1 day and 7 days.
Figure 11
Figure 11
The effect of Gelam honey on the expression of IL-6 protein in paw tissue. Rats were pretreated orally with Gelam honey (1 or 2 g/kg of body weight) for 1 or 7 days before inflammation was induced by carrageenan injection. D.W.: distilled water, Carr: carrageenan, IND: Indomethacin. Data are presented as the mean ± S.E.M. (n = 6). aP < 0.05 compared to the inflammation group (D.W. + Carr.). bP < 0.05 compared to the Indomethacin group (10 mg IND/kg of body weight + Carr.). cP < 0.05 compared between different honey doses at the same time point, 1 day or 7 days. dP < 0.05 the same honey dose, compared at different time points, 1 day and 7 days.

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