[논문]흰쥐에서 MIA로 유발된 골관절염에 선방활명음 가미방이 미치는 영향

신유빈; 박한별; 김재수; 이현종; 임성철; 이윤규

doi:10.14406/acu.2022.025

흰쥐에서 MIA로 유발된 골관절염에 선방활명음 가미방이 미치는 영향
Effects of Sunbanghwalmyung-Eum Gamibang on MIA-Induced Osteoarthritis in Rats 원문보기

Korean journal of acupuncture, v.39 no.4, 2022년, pp.152 - 171

신유빈 (대구한의대학교 한의과대학 침구의학교실) , 박한별 (대구한의대학교 한의과대학 침구의학교실) , 김재수 (대구한의대학교 한의과대학 침구의학교실) , 이현종 (대구한의대학교 한의과대학 침구의학교실) , 임성철 (대구한의대학교 한의과대학 침구의학교실) , 이윤규 (대구한의대학교 한의과대학 침구의학교실)

Abstract ▼ AI-Helper

Objectives : This study was designed to investigate the effects of Sunbanghwalmyung-eum gamibang on Monosodium iodoacetate-induced osteoarthritis rats. Methods : Forty Sprague-Dawley (SD) rats were divided into 5 groups of 8 rats each. Osteoarthritis (OA) was induced by injecting MIA (2 mg/50 µl) into the joint cavity of the left knee of SD rats belonging to the experimental group, and normal saline was injected into the joint cavity of the left knee instead of MIA in the normal group. To the normal group and the controlled group (OA group), 2 ml of distilled water was orally administered. To the positive control group (Indomethacin group), indomethacin 2 ml at a concentration of 2 mg/kg, to the low concentration group of SHG (Low group), 2 ml of SHG at a concentration of 2 mg/kg, and to the high concentration group of SHG (High group), 2 ml of SHG at a concentration of 4 mg/kg ml was orally administered. The drug was administered for a total of 4 weeks, and histological changes were analyzed by Hematoxylin-Eosin staining and Safranin-O staining. In addition, inflammatory cytokines such as TNF-α, IL-1β, and IL-6, and MMP-13, TIMP-1, and GAGs were immunohistochemically analyzed. Finally, hematological examination, blood biochemical examination, and liver and kidney biopsy were performed. Results : SHG groups (Low and High) inhibited the matrix destruction and damage of the knee joint cartilage in SD rat model, and significantly prevented the reduction in cartilage thickness. In SHG groups, the expressions of TNF-α, IL-1β, IL-6 and MMP-13 were significantly decreased, and the expressions of TIMP-1, GAGs were significantly increased compared with OA group. The safety indicators had no significant differences among five groups. Conclusions : These results show that SHG has cartilage protection capacity, anti-inflammatory effect.

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표/그림 (23)

표 Table 1. The composition and amount of Sunbanghwalmyung- eum gamibang
표 Table 2. Body weight changes of rats for 4 weeks
그림 Fig. 1. Comparisons of gross appearance of femur plateau in the rats. Normal, normal saline (injection at left knee)+distilled water(oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 2. Comparisons of gross appearance of tibial plateau in the rats. Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 3. The histological analyses of cartilage and bone changes in the rats. Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 4. Quantitative assessment of cartilage thickness in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^##p<0.01 compared with Normal group. *p<0.05, **p<0.01 compared with OA group. ^††p<0.01 compared with Indomethacin group. ^＄＄p<0.01 compared with Low group. Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 5. Articular cartilage of the rats knee with safranin-O staining. Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 6. Representative photographs of TNF-α immunoactivities of articular cartilage in the rats. TNF-α: tumor necrosis factor-α; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 7. Quantitative assessment of TNF-α immunohistochemistry in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^##p<0.01 compared with Normal group. *p<0.05, **p<0.01 compared with OA group.^††p<0.01 compared with Indomethacin group. TNF-α, tumor necrosis factor-α, Normal : normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 8. Representative photographs of IL-1β immunoactivities of articular cartilage in the rats. IL-1β, interleukin-1β; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 9. Quantitative assessment of IL-1β immunohistochemistry in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^##p<0.01 compared with Normal group. *p<0.05, **p<0.01 compared with OA group. ^††p<0.01 compared with Indomethacin group. ^＄p<0.05 compared with Low group. IL-1β, interleukin-1β, Normal, normal saline (injection at left knee)+distilled water (oral administration), OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 10. Representative photographs of IL-6 immunoactivities of articular cartilage in the rats. IL-6, interleukin-6; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+ distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 11. Quantitative assessment of IL-6 immunohistochemistry in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^#p<0.05, ^##p<0.01 compared with Normal group. *p<0.05, **p<0.01 compared with OA group. ^†p<0.05, ^††p<0.01 compared with Indomethacin group. IL-6, interleukin-6; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 12. Representative photographs of MMP-13 immunoactivities of articular cartilage in the rats. MMP-13, matrix metalloproteinase-13; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 13. Quantitative assessment of MMP-13 immunohistochemistry in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^##p<0.01 compared with Normal group. *p<0.05, **<0.01 compared with OA group. ^††p<,0.01 compared with Indomethacin group. MMP-13, matrix metalloproteinase-13; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 14. Representative photographs of TIMP-1 immunoactivities of articular cartilage in the rats. TIMP-1, tissue inhibitor of metalloproteinase-1; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 15. Quantitative assessment of TIMP-1 immunohistochemistry in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^##p<0.01 compared with Normal group. **p<0.01 compared with OA group. ^††p<0.01 compared with Indomethacin group. ^＄p<0.05 compared with Low group. TIMP-1, tissue inhibitor of metalloproteinase- 1; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 16. Representative photographs of GAGs immunoactivities of articular cartilage in the rats. GAGs, Glycosaminoglycans; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
그림 Fig. 17. Quantitative assessment of GAGs immunohistochemistry in the rats. Data represent the mean±standard deviation. Data were checked for significance using ANOVA and Dunnett's multiple comparison test to verify the significance levels of p<0.05 and p<0.01, respectively. ^##p<0.01 compared with Normal group. **p<0.01 compared with OA group. ^††p<0.01 compared with Indomethacin group. ^＄p<0.05 compared with Low group. GAGs, glycosaminoglycans; Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
표 Table 3. The Hematological Values of Rats
그림 Fig. 18. Hematoxylin & Eosin stained liver tissues from rats. Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).
표 Table 4. The serum biochemical values of rats
그림 Fig. 19. Hematoxylin & Eosin stained kidney tissues from rats. Normal, normal saline (injection at left knee)+distilled water (oral administration); OA, MIA (injection at left knee)+distilled water (oral administration); Indomethacin, MIA (injection at left knee)+indomethacin 2 mg/kg (oral administration); Low, MIA (injection at left knee)+2 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration); High, MIA (injection at left knee)+4 mg/kg of Sunbanghwalmyung-eum gamibang (oral administration).

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