Nutr Res Pract.  2016 Jun;10(3):265-273. 10.4162/nrp.2016.10.3.265.

Effect of Hijikia fusiforme extracts on degenerative osteoarthritis in vitro and in vivo models

• Kwon HO
• Lee M
• Kim OK
• Ha Y
• Jun W
• Lee J

• KMID: 2342128
• DOI: http://doi.org/10.4162/nrp.2016.10.3.265

Abstract

BACKGROUND/OBJECTIVES
The inhibitory effect of Hijikia fusiforme (HF) extracts on degenerative osteoarthritis was examined in primary cultured rat cartilage cells and a monosodium iodoacetate (MIA)-induced osteoarthritis rat model.
MATERIALS/METHODS
In vitro, cell survival and the expression of matrix metalloproteinases (MMPs), collagen type I, collagen type II, aggrecan, and tissue inhibitor of metalloproteinases (TIMPs) was measured after H2O2 (800 µM, 2 hr) treatment in primary chondrocytes. In vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats, and then RH500, HFE250 and HFE500 were administered orally once a day for 28 days. To determine the anti-inflammatory effects of HFE, nitric oxide (NO), prostaglandin E2 (PGE2) expression were measured. In addition, real-time PCR was performed to measure the genetic expression of MMPs, collagen type I, collagen type II, aggrecan, and TIMPs.
RESULTS
In the in vitro assay, cell survival after H2O2 treatment was increased by HFE extract (20% EtOH). In addition, anabolic factors (genetic expression of collagen type I, II, and aggrecan) were increased by HFE extract (20% EtOH). However, the genetic expression of MMP-3 and 7, known as catabolic factors were significantly inhibited by treatment with HFE extract (20% EtOH). In the in vivo assay, anabolic factors (genetic expression of collagen type I, II, aggrecan, and TIMPs) were increased by oral administration of HFE extract. However, the genetic expression of MMP-3 and 7, known as catabolic factors, and production of NO and PGE2 were significantly inhibited by treatment with oral administration of HFE extract.
CONCLUSIONS
HFE extract inhibited articular cartilage degeneration through preventing extracellular matrix degradation and chondrocyte injury.

Keyword

Chondrocyte; Collagen; MMP; TIMP

MeSH Terms

Administration, Oral
Aggrecans
Animals
Cartilage
Cartilage, Articular
Cell Survival
Chondrocytes
Collagen
Collagen Type I
Collagen Type II
Dinoprostone
Extracellular Matrix
In Vitro Techniques*
Injections, Intra-Articular
Knee Joint
Matrix Metalloproteinases
Models, Animal
Nitric Oxide
Osteoarthritis*
Rats
Real-Time Polymerase Chain Reaction
Tissue Inhibitor of Metalloproteinases
Aggrecans
Collagen
Collagen Type I
Collagen Type II
Dinoprostone
Matrix Metalloproteinases
Nitric Oxide
Tissue Inhibitor of Metalloproteinases

Figure

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  Fig. 1 Cell viability of primary cultured rat cartilage cells, determined by MTT reagent, with treatment of HF extracts at various concentrations. (A) Hot water extract 100-1,000 µg/mL, (B) 20% EtOH extract 100-1,000 µg/mL, (C) 50% EtOH extract 100-1,000 µg/mL. Data are expressed as mean ± SD (n = 3). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 2 Effect of HF extracts on survival against 800 µM H2O2 treatment. (A) Hot water extract 100-1,000 µg/mL, (B) 20% EtOH extract 100-1,000 µg/mL, (C) 50% EtOH extract 100-1,000 µg/mL. Data are expressed as mean ± SD (n = 3). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 3 Effect of HF extracts on genetic expression of cytokines in primary cultured cells. (A) TNF-α, (B) IL-6. HW; Hot water extract 100-1,000 µg/mL, ET20; 20% EtOH extract 100-1,000 µg/mL, ET50; 50% EtOH extract 100-1,000 µg/mL. Data are expressed as mean ± SD (n = 3). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 4 Effect of HF extracts on genetic expression of anabolic factors in primary cultured cells. (A) Col-1, (B) Col-2, (C) Aggrecan, (D) TIMP-1, (E) TIMP-3. HW; Hot water extract 100-1,000 µg/mL, ET20; 20% EtOH extract 100-1,000 µg/mL, ET50; 50% EtOH extract 100-1,000 µg/mL. Data are expressed as mean ± SD (n = 8/group). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 5 Effect of HF extracts on genetic expression of catabolic factors in primary cultured cells. (A) MMP-3, (B) MMP-7. HW; Hot water extract 100-1,000 µg/mL, ET20; 20% EtOH extract 100-1,000 µg/mL, ET50; 50% EtOH extract 100-1,000 µg/mL. Data are expressed as mean ± SD (n = 8/group). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 6 Inhibitory effects of HFE on serum NO level. Normal control; Not injection + AIN 93G, Sham; Injection MIA + AIN 93G, RH500; Injection MIA + AIN 93G + Rose Hip extract 500 µg/kg, HFE 250; Hijikia fusiforme 20% EtOH extract 250 µg/kg, HFE 500; Hijikia fusiforme 20% EtOH extract 500 µg/kg. Data are expressed as mean ± SD (n = 8/group). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 7 Inhibitory effects of HFE on serum PGE2 level. Normal control; Not injection + AIN 93G, Sham; Injection MIA + AIN 93G, RH500; Injection MIA + AIN 93G + Rose Hip extract 500 µg/kg, HFE 250; Hijikia fusiforme 20% EtOH extract 250 µg/kg, HFE 500; Hijikia fusiforme 20% EtOH extract 500 µg/kg. Data are expressed as mean ± SD (n = 8/group). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 8 Genetic expression of anabolic factors in MIA rat articular cartilage. Normal control; Not injection + AIN 93G, Sham; Injection MIA + AIN 93G, RH500; Injection MIA + AIN 93G + Rose Hip extract 500 µg/kg, HFE 250; Hijikia fusiforme 20% EtOH extract 250 µg/kg, HFE 500; Hijikia fusiforme 20% EtOH extract 500 µg/kg. Data are expressed as mean ± SD (n = 8/group). Significant differences were determined using Duncan's multiple range test at P < 0.05.

• 

  Fig. 9 Genetic expression of catabolic factors in MIA rat articular cartilage. Normal control; Not injection + AIN 93G, Sham; Injection MIA + AIN 93G, RH500; Injection MIA + AIN 93G + Rose Hip extract 500 µg/kg, HFE 250; Hijikia fusiforme 20% EtOH extract 250 µg/kg, HFE 500; Hijikia fusiforme 20% EtOH extract 500 µg/kg. Data are expressed as mean ± SD (n = 8/group). Significant differences were determined using Duncan's multiple range test at P < 0.05.

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