In Vitro and In Vivo Anti-Osteoarthritis Effects of 2,3,5,4'-Tetrahydroxystilbene-2-O-β-d-Glucoside from Polygonum Multiflorum

Abstract

Polygonum multiflorum Thunb. is a traditional herbal medicine that is rich in polyphenols. The major compound, 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside (THSG) has many pharmacological activities, such as antioxidative and free radical-scavenging properties, and the abilities to reduce hyperlipidemia, prevent lipid peroxidation, and protect the cardiovascular system. In this study, the anti-osteoarthritis (OA) effects of THSG were explored using in vitro and in vivo models. THSG inhibited nitric oxide (NO) and prostaglandin E₂ (PGE₂) production and inducible NO synthase (iNOS) and cyclooxygenase-2 expressions by lipopolysaccharide-stimulated RAW 264.7 cells. On the other hand, THSG inhibited PGE₂ production and iNOS and matrix metalloproteinase-13 expressions by interleukin-1β-stimulated primary rat chondrocytes. Through a mono-iodoacetate-induced rat OA model assay, THSG reduced paw edema and improved the weight-bearing distribution. Therefore, THSG has anti-inflammatory activity and could be applied as a lead compound for the development as an OA drug.

Keywords: ">d-glucoside; 2,3,5,4′-tetrahydroxystilbene-2-O-β-; Polygonum multiflorum; anti-inflammation; osteoarthritis.

Figure 1

Chemical structure of 2,3,5,4′-Tetrahydroxy-stilbene-2-O-β-d-glucoside (THSG).

Figure 2

The isolation flowchart of 2,3,5,4′-Tetrahydroxy-stilbene-2-O-β-d-glucoside (THSG) from Polygonum multiflorum Thunb. (PM).

Figure 3

The HPLC chromatogram of 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside. Column: LiChrospher 100 RP-18e (4 mm × 250 mm, 5 μm); Mobile phase: 0.05%TFA-CH₃CN (83:17); Flow rate: 1.0 mL/min; Column temperature: 40 ℃; Detection: 320 nm.

Figure 4

Anti-inflammatory of 2,3,5,4′-Tetrahydroxy-stilbene-2-O-β-d-glucoside (THSG) on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells after treatment for 6 h. iNOS and COX-2 protein expressions (A); ■ NO production inhibition rate of THSG and ☐ cell viability (B); PGE₂ production level (C), #: control group as compared to blank group, ### p < 0.005. *: TSHG group compared to control group, * p < 0.05, *** p < 0.005.

Figure 5

Effects of 2,3,5,4′-Tetrahydroxy-stilbene-2-O-β-d-glucoside THSG inhibited iNOS and MMP-13 expression on IL-β induced primary rat chondrocytes (A); PGE₂ production level (B), #: control group compared to blank group, ### p < 0.005; *: THSG group compared to control group, * p < 0.05, *** p < 0.005.

Figure 6

Paw edema volume of rats on mono-iodoacetate (MIA) injection in ankle of rat at 2nd day. Positive control (PC) is indomethacin (2.5 mg/kg). #: control group compared to blank group, ### p < 0.005. *: sample group as compared to control group, * p < 0.05, ** p < 0.001, *** p < 0.005.

Figure 7

Hind-limb weight bearing ratio of rats on mono-iodoacetate (MIA)-induced osteoarthritis (day 7–day 0). #: control group compared to blank group, ### p < 0.005. *: sample group compared to control group, *** p < 0.005.

Figure 8

The experimental schedule of mono-iodoacetate (MIA)-induced osteoarthritis model. MIA: first time injected 80 mg/mL MIA, 80 μL. (A) Paw edema measurement; (B) Hind-limb weighting bearing measurement.