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논문 기본 정보
- 자료유형
- 학위논문
- 저자정보
- 발행연도
- 2014
- 저작권
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Objectives :
This study was carried out to find out the anti-osteoarthritic effects of Mahwangbujaseshin-tang on the monosodium iodoacetate(MIA)-induced osteoarthritis SD-Rats.
Methods :
Osteoarthritis was induced by injecting MIA(50 ㎕) into the knee joint of rats. Rats were divided into a 3 groups(n=7). The injection did not fit the normal group. A week later, after the injection of MIA, while control group took normal saline 2 ㎖, the extract of Mahwangbujaseshin-tang (MBST)(200 ㎎/㎏) was injected to treated group. After that, we examined HPLC chromatogram, cytotoxicity, DPPH and ABTS free radical scavenging activity, ROS(reactive oxygen species) production, NO(Total Nitric oxide) production, Intracellular IL-1β(interleukin-1β), IL-6(interleukin-6), TNF-α, hind paw weight bearing ability, functions of liver and kidney, serum TNF-α, IL-1β, IL-6, PGE2, LTB4, TIMP-1, MMP-9 and hematology. Volume of cartilage was measured by micro CT arthrography. Injury of synovial tissue was measured by H & E, Safranin-O immunofluorescence.
Results :
1. The cytotoxicity against RAW 264.7 cells was disregardable and heavy metals were not detected or less than the reference value.
2. DPPH and ABTS free radical scavenging activity of MBST was increased according to concentration of MBST and total phenolic contents were in high level.
<in vitro>
1. ROS production was significantly decreased in MBST(at 10, 100 ㎍/㎖) and NO was also decreased but meaningless in MBST(at 100 ㎍/㎖).
2. IL-6 production was significantly decreased in MBST(at 100 ㎍/㎖) and TNF-α and IL-1β production were also decreased but meaningless in MBST(at 100 ㎍/㎖).
<in vivo>
1. In hind legs weight-bearing measurement, level of weight was increased.
2. ALT, AST, BUN and creatinine level in serum were decreased.
3. TNF-α, IL-1β and IL-6 production were significantly decreased.
4. PGE2, LTB4, MMP-9 and TIMP-1 production were significantly decreased.
5. In hematology, the levels of neutrophils, monocytes were significantly decreased and the levels of white blood cells, lymphocytes were also decreased but meaningless.
6. In micro CT-arthrography, cartilage volume was significantly increased.
7. Histopathologically, injury on cartilage and synovial membrane of MBST group was decreased.
Conclusions :
According to this study, Mahwangbujaseshin-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Mahwangbujaseshin-tang has anti-oxidants and anti-inflammation effects. And it is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.
Key words : Mahwangbujaseshin-tang, Osteoarthritis, Anti-inflammation, Anti-oxidation, MIA-induced.
This study was carried out to find out the anti-osteoarthritic effects of Mahwangbujaseshin-tang on the monosodium iodoacetate(MIA)-induced osteoarthritis SD-Rats.
Methods :
Osteoarthritis was induced by injecting MIA(50 ㎕) into the knee joint of rats. Rats were divided into a 3 groups(n=7). The injection did not fit the normal group. A week later, after the injection of MIA, while control group took normal saline 2 ㎖, the extract of Mahwangbujaseshin-tang (MBST)(200 ㎎/㎏) was injected to treated group. After that, we examined HPLC chromatogram, cytotoxicity, DPPH and ABTS free radical scavenging activity, ROS(reactive oxygen species) production, NO(Total Nitric oxide) production, Intracellular IL-1β(interleukin-1β), IL-6(interleukin-6), TNF-α, hind paw weight bearing ability, functions of liver and kidney, serum TNF-α, IL-1β, IL-6, PGE2, LTB4, TIMP-1, MMP-9 and hematology. Volume of cartilage was measured by micro CT arthrography. Injury of synovial tissue was measured by H & E, Safranin-O immunofluorescence.
Results :
1. The cytotoxicity against RAW 264.7 cells was disregardable and heavy metals were not detected or less than the reference value.
2. DPPH and ABTS free radical scavenging activity of MBST was increased according to concentration of MBST and total phenolic contents were in high level.
<in vitro>
1. ROS production was significantly decreased in MBST(at 10, 100 ㎍/㎖) and NO was also decreased but meaningless in MBST(at 100 ㎍/㎖).
2. IL-6 production was significantly decreased in MBST(at 100 ㎍/㎖) and TNF-α and IL-1β production were also decreased but meaningless in MBST(at 100 ㎍/㎖).
<in vivo>
1. In hind legs weight-bearing measurement, level of weight was increased.
2. ALT, AST, BUN and creatinine level in serum were decreased.
3. TNF-α, IL-1β and IL-6 production were significantly decreased.
4. PGE2, LTB4, MMP-9 and TIMP-1 production were significantly decreased.
5. In hematology, the levels of neutrophils, monocytes were significantly decreased and the levels of white blood cells, lymphocytes were also decreased but meaningless.
6. In micro CT-arthrography, cartilage volume was significantly increased.
7. Histopathologically, injury on cartilage and synovial membrane of MBST group was decreased.
Conclusions :
According to this study, Mahwangbujaseshin-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Mahwangbujaseshin-tang has anti-oxidants and anti-inflammation effects. And it is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.
Key words : Mahwangbujaseshin-tang, Osteoarthritis, Anti-inflammation, Anti-oxidation, MIA-induced.
목차
Ⅰ. 서 론 1Ⅱ. 실 험 31. 재료 및 방법 31) 재 료 3(1) 약재 3(2) 세포 3(3) 동물 3(4) 시약 4(5) 기기 52) 방 법 6(1) 시료 추출 6(2) HPLC 분석 6(3) 중금속 검사 6(4) 총 폴리페놀 함량 72. in vitro 및 in vivo 71) in vitro 7(1) 세포독성 측정 7(2) 항산화 활성 측정 8① DPPH 소거능 8② ABTS 소거능 8③ ROS 생성 9(3) 항염증 효능 측정 9① NO 생성 9② 염증관련 cytokine 분석 92) in vivo 10(1) MIA 주사에 의한 골관절염 유발 10(2) 군 분리 및 약물 투여 10(3) 뒷다리 체중 부하 검사 11(4) 혈액분석 11(5) 혈청 내 염증 cytokine 및 염증 매개 인자 측정 12① TNF-α, IL-1β, IL-6 12② Prostaglandin E2 12③ Leukotriene B4 13④ Matrix metalloproteinase-9 13⑤ Tissue inhibitor of metallopreteinases-1 13(6) 연골량 측정 14(7) 병리조직학적 검사 143. 통계처리 14Ⅲ. 결 과 151. HPLC 패턴 152. 중금속 함량 163. 총 폴리페놀 함량 16in vitro1. 세포 독성에 미치는 영향 172. 항산화 활성에 미치는 영향 181) DPPH 소거능 182) ABTS 소거능 193) ROS 생성 203. 항염증 효능에 미치는 영향 211) NO 생성 212) 염증 cytokine 생성 22(1) TNF-α 22(2) IL-1β 23(3) IL-6 24in vivo1. 뒷다리 체중 부하에 미치는 영향 252. 간 기능에 미치는 영향 263. 신장 기능에 미치는 영향 274. 염증 cytokine 및 염증 매개 인자에 미치는 영향 281) TNF-α 282) IL-1β 293) IL-6 304) Prostaglandin E2 315) Leukotriene B4 326) Matrix metalloproteinase-9 337) Tissue inhibitor of metallopreteinases-1 345. 혈액 내 면역세포에 미치는 영향 351) 총 백혈구 생성량 352) 호중구 생성량 363) 림프구 생성량 374) 단핵구 생성량 386. 연골량에 미치는 영향 397. 병리조직학적 변화에 미치는 영향 411) Hematoxylin & Eosin 염색(H & E) 412) Safranin-O 염색 42Ⅳ. 고 찰 43Ⅴ. 결 론 52참고문헌 54Abstract 61
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