Antinociceptive and Anti-Inflammatory Effects of Zerumbone against Mono-Iodoacetate-Induced Arthritis

Abstract

The fresh rhizome of Zingiber zerumbet Smith (Zingiberaceae) is used as a food flavoring and also serves as a folk medicine as an antipyretic and for analgesics in Taiwan. Zerumbone, a monocyclic sesquiterpene was isolated from the rhizome of Z. zerumbet and is the major active compound. In this study, the anti-inflammatory and antinociceptive effects of zerumbone on arthritis were explored using in vitro and in vivo models. Results showed that zerumbone inhibited inducible nitric oxide (NO) synthase (iNOS), cyclooxygenase (COX)-2 expressions, and NO and prostaglandin E₂ (PGE₂) production, but induced heme oxygenase (HO)-1 expression in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. When zerumbone was co-treated with an HO-1 inhibitor (tin protoporphyrin (SnPP)), the NO inhibitory effects of zerumbone were recovered. The above results suggest that zerumbone inhibited iNOS and COX-2 through induction of the HO-1 pathway. Moreover, matrix metalloproteinase (MMP)-13 and COX-2 expressions of interleukin (IL)-1β-stimulated primary rat chondrocytes were inhibited by zerumbone. In an in vivo assay, an acetic acid-induced writhing response in mice was significantly reduced by treatment with zerumbone. Furthermore, zerumbone reduced paw edema and the pain response in a mono-iodoacetate (MIA)-induced rat osteoarthritis model. Therefore, we suggest that zerumbone possesses anti-inflammatory and antinociceptive effects which indicate zerumbone could be a potential candidate for osteoarthritis treatment.

Keywords: Zingiber zerumbet Smith; anti-inflammatory; arthritis; heme oxygenase-1; metalloproteinase; zerumbone.

Figure 1

Anti-inflammatory action of zerumbone on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells after treatment for 6 h. (A) Inducible nitric oxide (NO) synthase (iNOS), cyclooxygenase (COX)-2, and heme oxygenase (HO)-1 protein expressions; (B) Quantitational and statistical analysis of protein expressions, * p < 0.05 compared to control; (C) prostaglandin E₂ (PGE₂) production level, * p < 0.05, the zerumbone group compared to the control group; (D) NO production inhibition of zerumbone co-treated with tin protoporphyrin (SnPP), * p < 0.05, the zerumbone group compared to the group co-treated with SnPP. C: control, cells were pretreated with vehicle and LPS (500 ng/mL) B: blank, the cells incubated with vehicle alone. Data are summarized and expressed as Mean ± SEM of three individual experiments (n = 3).

Figure 2

(A) Effects of zerumbone inhibiting matrix metalloproteinase (MMP)-13 expressions on interleukin (IL)-β-induced chondrocytes; (B) Quantitational and statistical analysis of protein expressions, * p < 0.05 compared to control; Data were used from three separate experiments; a picture of one is shown. C: control, cells were pretreated with vehicle and IL-1β (10 ng/mL) B: blank.

Figure 3

Abdominal writhing response of acetic-acid induces in mice. There were six mice per group. Results are expressed as Mean ± SEM. The positive control (PC) was morphine (5 mg/kg). * p < 0.05, compared to the control group.

Figure 4

Change in the swelling volume due to paw edema after a mono-iodoacetate (MIA) injection in the ankle of a rat on days 1 to 4. The positive control (PC) was indomethacin (10 mg/kg). *** p < 0.0005 compared to the control.

Figure 5

Distribution ratio of right and left hind-limb weight-bearing with mono-iodoacetate (MIA)-induced rat osteoarthritis on day 6. The positive control (PC) was indomethacin (10 mg/kg). * p < 0.05, *** p < 0.0005 compared to the control group.

Figure 6

Chemical structure of zerumbone isolated from the fresh rhizome of Zingiber zerumbet.

Figure 7

The experimental schedule of MIA-induced osteoarthritis model.