학술논문
제 II형 콜라겐으로 유도한 관절염 마우스에서 빈포세틴의 NLRP3 인플라마솜 억제에 의한 골미란 및 염증 완화 효과 : 콜라겐 유도 관절염 마우스에서 빈포세틴의 치료 효과 / Vinpocetine alleviates bone erosion and inflammation in type II collagen-induced arthritis in DBA1/J mice by suppressing NLRP3 inflammasome
Document Type
Dissertation/ Thesis
Author
Source
Subject
Language
English
Abstract
알칼로이드 빈카민의 반합성 유도체인 빈포세틴 (vinpocetine; VIN)은 항염증 효과가 있으며 장기간 사용 시 부작용이 거의 없는 것으로 널리 알려져 있다. 최근 연구에서는 NLRP3 인플라마솜이 류마티스 관절염 (RA)의 발병을 매개하는 중요한 단백질 복합체라고 알려지기 시작했다. 따라서, 본 연구는 VIN이 NLRP3 인플라마솜 발현 억제를 통해 RA의 증상을 완화하는지 조사하였다.콜라겐 유도 관절염 (CIA) 마우스는 0일차에 소 유래 제 2형 콜라겐 (BCII)과 Complete Freund’s adjuvant (CIA)로 면역화하고 21일차에 BCII와 Incomplete Freund’s adjuvant (IFA)로 활성화하여 유도되었다. RA의 특징적인 증상을 보고자 CIA 마우스의 발 두께 변화, 관절염 점수, 뒷발 회피 반응 역치 및 악력을 조사하였다. CIA 마우스의 뒷발과 무릎의 방사선 검사, 마이크로 컴퓨터단층촬영 및 조직적 평가는 RA의 병리학적 증상을 분석하는데 사용되었다. 실시간 정량적 역전사 중합 효소 연쇄 반응 (Real-time qPCR)과 웨스턴 블롯 분석은 NLRP3, 인터루킨 (IL)-1β, IL-18, 기타 염증성 사이토카인 그리고 NF-κB 리간드의 수용체 활성화 인자 (RANKL)의 발현을 확인하는 데 사용되었다. 효소면역측정법 (ELISA)은 CIA 마우스의 혈청에서 IL-18의 단백질 수준을 측정하기 위해 사용되었다.CIA 그룹과 비교하여 VIN 5 와 10 mg/kg은 CIA 마우스에서 관절 부종이나 RA 통증 평가로서의 악력 감소 및 기계적 유해자극에 대한 회피반응 등과 같은 RA의 특징적인 증상을 유의하게 완화시켰다. CIA 마우스의 골미란, 연골 파괴, 활액 과형성 등과 같은 RA의 병리학적 증상은 CIA+VIN 그룹 모두에서 개선되었다. 또한 RA 관련 염증성 사이토카인의 mRNA와 단백질 발현은 CIA 그룹에서 현저하게 증가했지만 CIA+VIN 그룹 모두에서 현저히 감소하였다. 특히, VIN은 CIA 마우스에서 NLRP3, IL-1β, IL-18과 같은 NLRP3 인플라마솜 인자의 발현을 유의적으로 억제하였다.결론적으로, VIN은 RA에서 중요 매개물질인 NLRP3 인플라마솜 관련 인자와 RA 관련 염증성 사이토카인의 발현을 억제함으로써 CIA 마우스의 골미란 및 염증을 완화시킬 수 있었으므로, 본 연구는 RA의 새로운 치료제로서 VIN의 사용에 대한 근거를 제공한다.
Vinpocetine (VIN), which is a semisynthetic derivative of the alkaloid vincamine, has been widely known to have anti-inflammatory effects and a rare side effect for long-term use. Currently, NLRP3 inflammasome has been known to be an important protein complex in rheumatoid arthritis (RA). This study investigated whether VIN improves the hallmark and pathological symptoms of RA, and reduces the expressions of RA-related inflammatory cytokines and NLRP3 inflammasome in the collagen- induced arthritis (CIA) model in DBA/1J mice. CIA model in DBA/1J mice was induced by immunization injection with Bovine type II collagen (BCII) and Complete Freund’s adjuvant (CFA) at day 0, and by boosting injection with BCII and Incomplete Freund's adjuvant (IFA) at day 21. The paw thickness, arthritis score, paw withdrawal threshold (PWT), and grip strength (GS) were investigated for the hallmark symptoms of RA in CIA mice. The X-ray, micro- CT, and histological analysis in the paw and knee were used to analyze the pathological symptoms of RA in CIA mice. The real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blotting were used to detect the expression of the RA-related inflammatory cytokines and NLRP3 inflammasome factors. The enzyme-linked immunosorbent assay (ELISA) was used to determine the protein level of IL-18 in the serum of CIA mice. In comparison to the CIA group, the paw swelling, PWT, and GS were dramatically relieved in the CIA+VIN 5mg/kg group and the CIA+VIN 10mg/kg group. The bone erosion, cartilage destruction, and synovial hyperplasia which are the pathological symptoms of RA were also improved in both VIN-treated groups compared with the CIA group. Moreover, mRNA and protein expressions of RA- related inflammatory cytokines were remarkably increased in the CIA group but these were significantly decreased in the VIN-treated groups. In particular, VIN dramatically inhibited the mRNA and protein expressions, and protein level of NLRP3 inflammasome factors such as NLRP3, IL-1β, and IL-18 in CIA mice. Taken together, VIN could alleviate bone erosion and inflammation in CIA mice by suppressing NLRP3 inflammasome, this study provides a novel insight that VIN might be used as a new therapeutic for the treatment of RA.
Vinpocetine (VIN), which is a semisynthetic derivative of the alkaloid vincamine, has been widely known to have anti-inflammatory effects and a rare side effect for long-term use. Currently, NLRP3 inflammasome has been known to be an important protein complex in rheumatoid arthritis (RA). This study investigated whether VIN improves the hallmark and pathological symptoms of RA, and reduces the expressions of RA-related inflammatory cytokines and NLRP3 inflammasome in the collagen- induced arthritis (CIA) model in DBA/1J mice. CIA model in DBA/1J mice was induced by immunization injection with Bovine type II collagen (BCII) and Complete Freund’s adjuvant (CFA) at day 0, and by boosting injection with BCII and Incomplete Freund's adjuvant (IFA) at day 21. The paw thickness, arthritis score, paw withdrawal threshold (PWT), and grip strength (GS) were investigated for the hallmark symptoms of RA in CIA mice. The X-ray, micro- CT, and histological analysis in the paw and knee were used to analyze the pathological symptoms of RA in CIA mice. The real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blotting were used to detect the expression of the RA-related inflammatory cytokines and NLRP3 inflammasome factors. The enzyme-linked immunosorbent assay (ELISA) was used to determine the protein level of IL-18 in the serum of CIA mice. In comparison to the CIA group, the paw swelling, PWT, and GS were dramatically relieved in the CIA+VIN 5mg/kg group and the CIA+VIN 10mg/kg group. The bone erosion, cartilage destruction, and synovial hyperplasia which are the pathological symptoms of RA were also improved in both VIN-treated groups compared with the CIA group. Moreover, mRNA and protein expressions of RA- related inflammatory cytokines were remarkably increased in the CIA group but these were significantly decreased in the VIN-treated groups. In particular, VIN dramatically inhibited the mRNA and protein expressions, and protein level of NLRP3 inflammasome factors such as NLRP3, IL-1β, and IL-18 in CIA mice. Taken together, VIN could alleviate bone erosion and inflammation in CIA mice by suppressing NLRP3 inflammasome, this study provides a novel insight that VIN might be used as a new therapeutic for the treatment of RA.