Abstract

Objectives :
This study was performed to investigate the effects of Bujasasim-tang ethanol extract(BST) on oxidative stress, inflammation and osteoarthritic rat model.

Methods :
To ensure safety of BST, heavy metal levels were measured and cytotoxicity test was done.
in vitro
To evaluate antioxidative effects of BST, total phenolic contents, 1,1-diphenyl-2-picryl-hydrazyl(DPPH), 2,2''?azino?bis-(3-ethylbenzothiazoline-
6-sulfonic acid)(ABTS) scavenging activity, reactive oxygen species(ROS) levels were measured. Also, to evaluate anti-inflammatory effects of BST treated group, total nitric oxide(NO) and pro-inflammatory cytokines(IL-1β, IL-6, TNF-α) levels were measured in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells.

in vivo
We injected MIA 50 ㎕(60 ㎎/㎖) into knee joints of rats to induce osteoarthritis. Rats were divided into total 3 groups(normal, control, BST treated group, each n=7). Normal group was not treated at all without inducing osteoarthritis and taken normal diet. Control group was induced osteoarthritis by MIA and taken with 2 ㎖ of distilled water once a day for 4 weeks. BST treated group was induced osteoarthritis by MIA and taken BST 2 ㎖(200 ㎎/㎏/mouse) once a day for 4 weeks.
We evaluated dynamic weight bearing with the Incapacitance Test Meter. At the end of experiment, the rats were sacrificed to observe the functions of liver and kidney, changes of WBC, neutrophil, lymphocyte, monocyte levels in blood, to evaluate the levels of pro-inflammatory cytokines, tissue inhibitor of metallopreteinases-1(TIMP-1), matrix metalloproteinase-9(MMP-9), prostaglan-
din E2(PGE2), leukotriene B4(LTB4) within serum. We observed change of articular structures by Hematoxylin & Eosin(H&E), safranin-O staining method and measured amount of cartilage by micro CT-arthrography. Statistical analysis was done by unpaired student''s t-test with significance level at p<0.05 in SPSS 11.0 for windows.

Results :
1. Safety of the BST was identified.
2. AST, ALT, BUN, creatinine levels of BST treated group were within normal limit.

in vitro
1. DPPH and ABTS free radical scavenging activities of BST showed dose-dependent increase.
2. ROS production were significantly decreased.
3. Total nitric oxide(NO) and IL-1β production were decreased.
4. IL-6 and TNF-α production were significantly decreased.

in vivo
1. Weight bearing ability was significantly increased.
2. WBC, neutrophil, lymphocyte, monocyte levels in blood were decreased.
3. IL-1β and TNF-α levels in serum were significantly decreased. and the IL-6 level was decreased.
4. TIMP-1, MMP-9, LTB4, PGE2 levels in serum were significantly decreased.
5. Cartilage volume of BST treated group was significantly increased. Also changes of cartilage, synovial membrane, fibrous tissue were suppressed.

Conclusions :
The results obtained in this study Bujasasim-tang have effects of antioxidative, anti-inflammatory, relieve pain and protection of cartilage. Therefore we expect that Bujasasim-tang is effective treatment for osteoarthritis.

Key words : Bujasasim-tang, Osteoarthritis, Oxidative stress, Anti-inflammation, Monosodium iodoacetate(MIA)