Evaluation of Apoptosis in Collagen-induced Arthritis with 123I-Annexin V Scintigraphy

碘－123-Annexin V造影檢查對膠原誘導關節炎自體凋亡之評估

李聖慧(Sheng-Huei Lee) ； 廖美秀(Mei-Hsiu Liao) ； 江昭志(Chao-Chih Chiang) ； 沈立漢(Lie-Hang Shen) ； 林萬鈺(Wan-Yu Lin)

《核子醫學雜誌》 22卷3期 (2009/09) Pp. 169-176

https://doi.org/10.6332/ANMS.2203.004

摘要

背景：風濕性關節炎是一種慢性自體免疫疾病，其主要特徵爲巨嗜細胞與T細胞不正常積聚於關節因而導致關節發炎。自體凋亡亦被認爲是此疾病之重要機轉。以放射藥物標記annexin V已被明對偵測自體凋亡相當有用。近來，國內核研所成功研發以碘－123標誌annexin V，本研究即以核研所所研發之碘－123 annexin V評估經膠原誘導關節炎之老鼠其自體凋亡之嚴重度。
方法：本研究使用10隻SD老鼠，分成關節炎老鼠組與正常對照組（每組5隻）。我們使用眞皮內注射type II膠原於老鼠背部和尾巴之方法以誘導關節炎。在經尾靜脈注射碘－123 annexin V後4小時以及24小時，所有老鼠均進行核醫造影。影像結果經電腦處理，於老鼠之左後踩關節與左復大腿軟組織分別圈定興趣區並計算足踝關節與大腿肌肉之活性比值(A/M ratio)。造影結束後，犠牲老鼠進行H&E、TUNEL以及DAPI染色檢查。
結果：在關節炎老鼠組可見老鼠關節明顯紅腫。此外，於造影中亦可見紅腫關節有明顯之碘－123標誌annexin V攝取。相反地，在正常老鼠組，沒有明顯之碘－123標誌annexin V攝取。在注射碘－123標誌annexin V後4小時以及24小時，關節炎老鼠組之A/M ratio比起正常老鼠組明顯較高。使用Mann-Whitney U test統計法分析其P值小於0.05。以TUNEL和DAPI方法染色可以見到在關節炎老鼠組之踩關節有自體凋亡之現象，但在正常老鼠組之踩關節則不見自體凋亡之現象。
結論：核研所研發之碘－123標誌annexin V在膠原誘導關節炎之老鼠可有效評估其關節自體凋亡之情形，未來應有潛力成爲一個可以體內評估人體自體凋亡之有效方法。

關鍵字

膠原誘導關節炎 ； 自體凋亡 ； 碘－123標誌annexin V

並列摘要

Background: Rheumatoid arthritis (RA) is a chronic progressive disease and an autoimmune disorder characterized by joint inflammation due to abnormal accumulation of macrophages and autoreactive T lymphocytes in joints. Apoptosis is considered to be part of the pathogenesis of the disease. The radiolabeled annexin V has been proven effective in detecting apoptotic cells. Recently, the Institute of Nuclear Energy Research (INER) has successfully labeled annexin V with 123I. In this study, we used INER-produced 123I-Annexin V for the evaluation of apoptotic status in collagen-induced arthritis (CIA) rats.
Methods: Ten Sprague-Dawley (SD) rats were divided into two groups: a control group and a CIA group (five in each group). Bovine type II collagen emulsified with complete Freund's adjuvant was injected intradermally at the back and tail to induce arthritis in SD rats. Scintigraphy was performed 4h and 24h after injection of 123I-annexin V in both CIA rats and control rats. ROI was selected for the ankle joint and the soft tissue of the thigh of the left rear leg in all rats. Activity of ankle to muscle (A/M ratio) was calculated by computer. Afterwards, rats were sacrificed for H&E, TUNEL and DAPI (diamidino-2-phenylindole) staining.
Results: Obvious swelling of the left rear legs was noted in CIA rats. Increased 123I-annexin V uptake in the ankles and paws was noted in the CIA rats. In contrast, no significant 123I-annexin V uptake was noted in the ankles of the control rats. The A/M ratios in CIA rats were significantly higher when compared to that in control rats at both 4h and 24h after 123I-annexin V injection. The P values were less than 0.05 using the Mann-Whitney U test. Using the TUNEL and DAPI stain, apoptotic cells were seen in the ankle joints of the left rear legs in CIA rats but not in the ankle joints of normal rats.
Conclusion: 123I-annexin V produced by INER can be used to evaluate apoptosis in the joints of a CIA rat and may be a potential agent for the in-vivo evaluation of apoptosis in humans.

並列關鍵字

collagen-induced arthritis ； apoptosis ； 123I-annexin V