The purpose of this study was to investigate the lesions of a mouse collagen antibody-induced arthritis (CAIA) model using fluorescence bioimaging and micro-computed tomography (micro-CT) and to compare it with histopathological examination. Twelve mice were randomly divided into three groups: group 1 (G1) as control, group 2 (G2) as fluorescence probe control and group 3 (G3) as collagen antibodyinduced arthritis. The mice of G3 intravenously received anti-type II collagen 5-clone antibody cocktail (2 mg/mouse) on day 0 and intraperitoneally received lipopolysaccharide (50 μg/mouse) on day 3. On the while, the mice of G1 and G2 received 0.9% saline in equal volumes at equivalent times. Fluorescence bioimaging and micro-CT analysis were carried out to assess arthritis. Treatment with the collagen antibody cocktail increased the paw thickness of mice compared to those in both the control and probe-treated groups. Fluorescence bioimaging using a near infrared imaging agent showed high intensity in the joints of collagen antibody- treated mice, whereas those of control mice showed no signal. Micro-CT analysis of the knee joints of collagen antibody-treated mice showed rough and irregular articular appearance, whereas those of control mice showed normal appearance. Histopathological examination of the knee joints of collagen antibody-treated mice revealed destruction of cartilage and bony structure, synovial hyperplasia and infiltration of inflammatory cells. No cartilage destruction or inflammation was observed in control or probe control mice. Taken together, it is concluded that analyses of fluorescent bioimaging made it possible to evaluate CAIA lesions, comparable with those by micro-CT and histopathological examination in mice.

Calcineurin-binding protein 1 (Cabin1) regulates calcineurin phosphatase activity as well as the activation, apoptosis, and inflammatory responses of fibroblast-like synoviocytes (FLSs), which actively participate in the chronic inflammatory responses in rheumatoid arthritis (RA). However, the mechanism of action of Cabin1 in FLS apoptosis is not clear. The aim of this study was to define the regulatory role of Cabin1 in FLSs of mice with collagen-induced arthritis (CIA). Transgenic mice overexpressing human Cabin1 in joint tissues, under the control of a type II collagen promoter, were generated. hCabin1 expression in joints and FLSs was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of cytokines, matrix metalloproteinases (MMPs), and apoptosis-related genes in FLSs was determined by enzyme- linked immunosorbent assay, gelatin zymography, and RT-PCR, respectively. Joints were histologically examined after H&E and TRAP staining. hCabin1-transgenic CIA mice had less severe arthritis than wild-type CIA mice, based on hind paw thickness and histology. This was accompanied by significantly enhanced apoptosis in transgenic mice, evidenced by significantly more TUNEL-positive cells in synovial tissues. The expression of inflammatory cytokines and MMPs was reduced, and the transgenic CIA mice exhibited decreased Akt activation and increased expression of p53, caspase-3, caspase-9, and Bax. hCabin1 plays a critical role in promoting apoptosis of FLSs and in attenuating inflammation and the destruction of cartilage and bone in RA. These findings help elucidate the pathogenic mechanisms of RA and suggest that Cabin1 is a potential target for RA treatment.

Background: Taraxacum platycarpum has been used in traditional medicine in Korea to treat intoxication and edema and as a diuretic. According to previous reports, it has anti-cancer, anti-gastritis, and anti-inflammation effects. However, the improvement effect of T. platycarpum on rheumatoid arthritis has not been investigated. The anti-oxidative and anti-inflammation effects of the aerial parts of T. platycarpum are different from those of its subterranean parts. Thus, we evaluated the effect of the water extracts of Taraxaci radix (WTR) on type II collagen-induced rheumatoid arthritis (CIA) in animal models. Methods and Results: Rheumatoid arthritis was induced by type II collagen. WTR (100㎎/㎏ and 500㎎/㎏) was administered to the animal models. Methotrexate was used as the positive control. The levels of interleukin-6, TNF-alpha, and type II collagen IgG in the animals were measured by using enzyme-linked immunosorbent assay. Treatment with 500㎎/㎏ WTR decreased the serum levels of interleukin-6, TNF-alpha, and collagen IgG in the CIA models. Moreover, treatment with WTR diminished the arthritisinduced swelling of the hind legs and monocyte infiltration in the bloodvessels of the animal models. Conclusions: These results indicate that WTR has the potential to improve rheumatoid arthritis by reducing the levels of inflammatory cytokines such as interleukin-6 and TNF-alpha. However, further experiments are required to elucidate the influence of WTR on signal transduction in vitro and in vivo.

류마티스성 관절염의 동물 모델인 CIA(collagen induced arthritisd) 생쥐에 CRN을 복강 투여하여 CIA의 발병률 및 관절염지수 등에 대한 효과를 조사한 결과는 다음과 같다. RF(rhumatoid factor) 인자인 IgG와 IgM 생산량, 그리고 염증 사이토카인인 IL-6와 TNF-α의 생산량이 감소하였다. 사이토카인 분석결과, 비장세포에서 Th1과 Th2 세포에서 분비되는 IFN-γ와 IL-4를 측정하여 CRN 투여로 Th1 세포에서 Th2 세포로 면역반응이 전환됨을 시사하는 것은 CRN의 치료효과로 사료된다. 혈청 중에서 RA 인자인 IgG와 IgM, 염증 사이토카인인 TNF-α와 IL-6 및 type II bovine collagen antibody 생산량 이 대조군에 비하여 CRN 투여군이 감소하여 CRN이 관절염의염증 사이토카인을 억제하는 것으로 확인된다.

본 연구에서는 택칠에서 분리된 CRN(corilagin)이 류마티스성 관절염 치료제로 사용될 수 있는가능성을 조사하기 위하여 류마티스성 관절염의 동물 모델인 CIA(collagen induced arthritis) 생쥐에 CRN을 복강 투여하여 CIA의 발병률 및 관절염 지수 등에 대한 효과를 관찰하여 다음과 같은 결과를 얻었다. 택칠에서 분리한 corilagin을 관절염 유발 생쥐에게 투여한 결과, 관절염의 발병진행이 억제되고 관절염 지수가 감소되는 것을 확인하였다. 세포 조직학적 관찰을 통하여 염증성 세포들이 현저하게 줄어들고, 관절강이 잘 확보되었으며, pannus의 형성이 보이지않고 연골 또한 잘 보호되어져 있음을 확인하였다. 관절염 유발 생쥐의 형광유세포 분석결과 각 조직에 침윤되는 염증세포가 감소하였다.