계명대학교 의학도서관 Repository

강직척추염에서 골 재형성, 염증에 대한 CFHR5 역할 규명

Metadata Downloads
Issued Date
2023-02
Abstract
Ankylosing spondylitis (AS) is a chronic inflammatory rheumatic disease in which the spine becomes inflamed. The exact etiology is unknown. In prior studies, it was confirmed that complement factor H-related 5 (CFHR5), a protein of the complement system, was upregulated in the synovial fluid of AS. This study aimed to investigate the effect of CFHR5 on bone remodeling and inflammation in AS in vitro and in vivo. Osteoblast differentiation and mineralization were observed when CFHR5 (0 – 1000 ng/ml) was added to an osteoblast cell line (MC3T3-E1 cells) and AS, control (Ct) osteoprogenitor cells. In SKG mice, an AS-induced animal model, the experiment was conducted with a control group (n = 5) and a CFHR5 group (n = 5) and CFHR5 (0.5 mg/kg) was administered twice for 5 weeks. To confirm bone formation and inflammation, micro-CT and histopathological analysis (H&E, Safranin O) were performed on the paws of mice. As a result, it was confirmed that alkaline phosphatase (ALP) expression in MC3T3-E1 cells, AS and Ct osteoprogenitor cells was highly expressed in the CFHR5-treated group, causing bone differentiation. Alizarin red (ARS) was also highly expressed in the CFHR5 treatment group and it was confirmed that it causes bone calcification. Expressions of runt-related transcription factor 2 (RUNX2) and osteopontin (OPN) were confirmed in MC3T3-E1 cells. After CFHR5 treatment, RUNX2 showed the strongest expression at 7 days and OPN at 3 days and there was no significant difference by concentration. In the SKG animal model, there was no significant difference in inflammatory activity between the control and CFHR5-treated groups. Micro-CT and histological analysis of the ankles of mice confirmed that bone formation significantly increased in the CFHR5-treated group. In this study, it was confirmed that CFHR5 causes bone remodeling in vitro and in vivo models of AS. The above results will be important clues to confirm the immunological mechanism between AS and complement system proteins, suggesting that CFHR5 can be a biomarker protein for AS.
강직척추염(Ankylosing Spondylitis, AS)은 척추에 염증이 발생하여 척추 마디가 굳어지는 만성 염증성 류마티스 질환으로, 명확한 병인은 밝혀지지 않았다. 선행 연구를 통해 AS의 활액에서 보체계 단백질인 complement factor H-related 5(CFHR5)가 과발현되는 것을 확인하였다. 이 연구의 목표는 in vitro 및 in vivo에서 CFHR5가 AS의 골 재형성 및 염증에 미치는 영향을 확인하는 것이다. 조골세포주(MC3T3-E1 cells) 및 AS, control(Ct) 골전구세포에서 CFHR5(0 – 1000 ng/ml)를 처리했을 때, 골 분화 및 석회화를 관찰하였다. AS 유도 동물 모델인 SKG 마우스에서 대조군(n=5), CFHR5군(n=5)으로 실험을 진행하였으며, CFHR5(0.5mg/kg)를 5주간 2회씩 투여하였다. 골 형성 및 염증을 확인하기 위해 쥐의 발에서 micro-CT와 조직병리학적 분석(H&E, Safranin O)을 수행하였다. 그 결과, MC3T3-E1 cells 및 AS, Ct 골전구세포에서 alkaline phosphatase(ALP)의 발현이 CFHR5 처리군에서 높게 발현되었으며, 골 분화를 일으키는 것을 확인하였다. Alizarin red(ARS) 역시 CFHR5 처리군에서 높게 발현되었으며, 골 석회화를 일으키는 것을 확인하였다. MC3T3-E1 cells에서 runt-related transcription factor 2(RUNX2), osteopontin(OPN)의 발현을 확인하였다. CFHR5 처리 후 RUNX2는 7일, OPN은 3일에서 가장 강한 발현을 보였으며, 농도 별로는 큰 차이를 보이지 않았다. SKG 동물 모델에서 대조군 및 CFHR5 처리군 사이의 염증 활성에는 큰 차이가 없었다. 쥐의 발목에서 micro-CT와 조직학적 분석을 확인하였을 때, CFHR5 처리군에서 골 형성이 현저히 증가한 것을 확인하였다. 이 연구에서는 CFHR5가 AS의 in vitro 및 in vivo 모델에서 골 재형성을 일으키는 것을 확인하였다. 위 결과는 AS와 보체계 단백질 사이의 면역학적 기전을 확인하는데 중요한 단서가 될 것이며, CFHR5가 AS의 바이오마커가 될 수 있음을 시사한다.
Alternative Title
The role of CFHR5 on bone remodeling and inflammation in ankylosing spondylitis
Awarded Date
2023-02
Degree
박사
Type
Thesis
URI
https://kumel.medlib.dsmc.or.kr/handle/2015.oak/45145
Appears in Collections:
1. School of Medicine (의과대학) > 박사
Authorize & License
  • Authorize공개
Files in This Item:

Items in Repository are protected by copyright, with all rights reserved, unless otherwise indicated.