• 혜화의학회지
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• 제24권2호
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• pp.35-57
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• 2016

Objectives : The purpose of this study is to prove the effect and mechanism of Gamikyejakjimogawusul-tang(GKHA) herbal acupuncture on induced rheumatoid arthritis model of DBA/1 mice. Methods : We check effect of GKHA extract on the AST, ALT, Creatinine, BUN of serum and cell viability of GK extract in RAW 264.7 cells to test the stability of this study. In vitro, we measure total phenol contents, total flavonoid contents, DPPH free radical scavenging activity, ABTS cation radical scavenging activity of Gamikyejakjimogawusul-tang, effect of GK extract on ROS(Reactive Ooxygen Species) production to estimate a anti-oxidant capacity, and we also measure effect of GK extract on NO (Nitric Oxid), IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF production in RAW 264.7 cells to estimate a anti-inflammatory efficacy. In vivo, we compare a rheumatoid arthritis manifestation between control and experimental group and estimate a AI. Then we check effect of GKHA on the level of WBC, neutrophil, lympocyte, monocyte in the blood to see the effect of immune cells in blood. In addition we measure effect of GKHA on the level of hs-CRP, IgM, IgG, IL-$1{\beta}$, IL-6, IL-17, IL-21, TNF-${\alpha}$, MCP-1, GM-CSF in serum. We observe effects of GKHA on imaging of cartilage degeneration using micro CT-arthrography in paw hind. And we calculate effects of GKHA that reduced BV ratio, BS/BV ratio using 3D Micro-CT. Lastly we observe effects of GKHA histopathologic examination analysis. Results : 1. The toxicity on liver and kidney was disregardable and the cytotoxicity against RAW 264.7 cells was also disregardable. 1. Total phenol contents and total flavonoid contents in GK extract were in high level. 2. DPPH free radical scavenging activity and ABTS cation radical scavenging activity were increased according to concentration of GK extract 3. ROS production was significantly decreased in GK extract (at 10, $100{\mu}g/ml$). 4. NO, IL-6, TNF-${\alpha}$, MCP-1 production were significantly decreased in GK extract(at 10, $100{\mu}g/ml$). IL-17, GM-CSF production were significantly decreased in GK extract(at 1, 10, $100{\mu}g/ml$). IL-$1{\beta}$, IL-21 production were also decreased but there was no statistical significance. 5. 25x observation after H&E and M-T staining, infiltration of immune cells and subsidence of the cartilage and damage to the synovial cells were decreased. Conclusions : This study showed that GKHA extract had anti-oxidant capacity, anti-inflammatory efficacy. GKHA extract also had inhibiting effect on the process of rheumatoid arthritis and can protect joint and cartilage. So we expect that GKHA extract can be a meaningful treatment to rheumatoid arthritis patients.

• Journal of Acupuncture Research
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• 제19권5호
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• pp.161-175
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• 2002

봉독은 거풍습(祛風濕), 지통(止痛), 해경평천(解痙平喘), 소종강압(消腫降壓)의 효능으로 오랫동안 통증과 염증성 질환을 치료하는데 이용되어져 왔는데 최근에는 면역관련질환치료에 응용하여 좋은 결과가 보고되고 있다. 본 연구는 Rheumatoid arthritis와 유사한 형태의 대표적 실험모델로 알려진 실험용 쥐의 Type II collagen 유발 관절염(Type II collagen induced arthritis : CIA)의 활액에서 봉독약침이 단백분해효소와 유리기 손상에 미치는 면역억제효과를 알아보기 위해 실행되었다. 본 실험에서는 CIA가 유발된 실험용 쥐에 봉독약침($5{\mu}l/kg$)을 처리한 실험군과 대조군으로서 CIA 유발 쥐에 생리식염수를 처리한 군(CIA군), 정상적인 쥐에 생리식염수로 처리한 군(정상군)으로 구분하여 각 군들의 일련의 표본에서 세포질, 리소좀, 간질성 단백분해효소의 활성과 유리기로 인한 단백질 손상정도를 (carbonyl 유도체를 측정하여)서로 비교하였다. 그 결과 각 군의 활액표본에서는 많은 종 류의 단백분해효소가 정산군보다 CIA군에서 유의하게 활성이 높았으며, 봉독약침($5{\mu}l/kg$)을 처리한 군에서 효소들의 활성이 유의하게 감소하였다. 그러나 각 군들의 혈장표본에서는 이 효소들의 활성은 서로 유의한 차이가 없었다. 이는 혈장속의 면역반응과 연관되리라고 추측되는 단백분해효소들의 활성변화는 병인적 측면에서 RA와 같은 염증성관절 질환과는 큰 상관성이 없다는 것을 의미한다. Carbonyl 유도체 측정으로 평가한 유리기 손상은 활액과 현장표본에서 모두 봉독약침($5{\mu}l/kg$)을 처리한 군에서 유의성 있게 감소하였다. 이상의 결과로 볼 때 단백분해효소와 유리기의 활성은 RA의 병인학적 측면에서 모두 잠재적인 중요성을 가지고 있으므로 향후 새로운 RA치료법은 이들 단백분해효소의 활성저해와 유리기의 소거능을 포함해야 한다고 사료되며 봉독약침은 이러한 2가지 효능을 포함한 효과적인 치료라고 평가된다.

• 대한한의학방제학회지
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• 제29권2호
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• pp.81-91
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• 2021

Objectives : This study was conducted to evaluate the efficacy of a complex mixture of natural substances of ginseng and baeknyeoncho on the arthritic rats. Methods : In vitro experiments were conducted to ensure the stability of the complex. After setting toxicity and concentration by MTT assay, the antioxidant effect was measured through DPPH and ABTS radical scavenging activity. To confirm the anti-inflammatory effects of the complex, levels of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, TNF-α) were measured in LPS-treated macrophage cell lines (RAW264.7). We injected monosodium iodoacetate (MIA) 50 μl (60 mg/ml) into knee joints of rats to induce osteoarthritis. The rats were divided into three groups (normal (n=5), control (n=5), and OR (n=5) group). The control group consumed 2 mg/kg of physiological saline once a day for 4 weeks, and the OR group was mixed at a concentration of 416.5 mg/kg of Baengnyeoncho (O) and 208.25 mg/kg of red ginseng (R) and ingested 1 mL each 5 days a week. Results : This complex increased the DPPH and ABTS radical scavenging rate. The complex decreased NO production and pro-inflammatory cytokine production of macrophages. In the OR group, the secretion of cytokine in serum was decreased. In histopathological examination, the joint tissue of the composite showed less damage to the synovial membrane, cartilage, and fibrous tissue than the control group. Conclusions : As a result of this study, natural complexes have antioxidant, anti-inflammatory and cartilage protection effects. Therefore, we expect the complex to be effective in treating osteoarthritis.

• IMMUNE NETWORK
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• 제22권4호
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• pp.34.1-34.19
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• 2022

Osteoarthritis (OA) is the most common form of arthritis associated with ageing. Vitamin D has diverse biological effect on bone and cartilage, and observational studies have suggested it potential benefit in OA progression and inflammation process. However, the effect of vitamin D on OA is still contradictory. Here, we investigated the therapeutic potential of vitamin D in OA. Six-week-old male Wistar rats were injected with monosodium iodoacetate (MIA) to induce OA. Pain severity, cartilage destruction, and inflammation were measured in MIA-induced OA rats. Autophagy activity and mitochondrial function were also measured. Vitamin-D (1,25(OH)₂D3) and celecoxib were used to treat MIA-induced OA rats and OA chondrocytes. Oral supplementation of vitamin D resulted in significant attenuations in OA pain, inflammation, and cartilage destruction. Interestingly, the expressions of MMP-13, IL-1β, and MCP-1 in synovial tissues were remarkably attenuated by vitamin D treatment, suggesting its potential to attenuate synovitis in OA. Vitamin D treatment in OA chondrocytes resulted in autophagy induction in human OA chondrocytes and increased expression of TFEB, but not LC3B, caspase-1 and -3, in inflamed synovium. Vitamin D and celecoxib showed a synergistic effect on antinociceptive and chondroprotective properties in vivo. Vitamin D showed the chondroprotective and antinociceptive property in OA rats. Autophagy induction by vitamin D treatment may be a promising treatment strategy in OA patients especially presenting vitamin D deficiency. Autophagy promoting strategy may attenuate OA progression through protecting cells from damage and inflammatory cell death.