한우의 혈액시료 처리 방법(시간, 온도) 따른 혈액 내 사이토카인 및 생화학적 변화
Blood Sample Handling Method (Time, Temperature) for Cytokines and Biochemical Analysis in Hanwoo
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혈청을 이용한 분석은 다양한 질병 진단의 지표로 이용된다. 혈청의 분리 방법, 저장시간 및 온도에서 부적절한 혈액 처리는 사이토카인의 농도와 생화학 검사결과에 영향을 줄 수 있다. 본 연구에서는 한우를 공시하여 혈청 분리 시기, 저장 시간과 온도가 사이토카인 및 생화학 검사결과에 미치는 영향을 조사 하였다. 혈액은 경정맥에서 채취하여, 26±3℃에서 1~2시간 방치 후 분리조건을 다음과 같이 4개의 그룹 (a-d)으로 나뉘어 진행하였다. a) 혈청 분리하여 보관(대조구), b) 혈청분리 후 아이스박스 2시간 방치 후 보관, c) 아이스박스 2시간 방치 후 혈청분리, d) 24시간 4℃ 방치 후 혈청분리, 이러한 조건으로 분리된 혈청은 분석 전까지 –70℃ 보관 하였다. 혈청 분리시기, 아이스박스 또는 4℃ 저장 시간 및 온도에 따라 IFN-γ는 유의적인 차이를 보이지 않았고, TNF-α의 농도는 그룹 a-c에 비해 그룹 d에서 유의적(p<0.05)으로 낮아졌다. 또한 albumin, total protein의 농도는 그룹 a보다 d에서 유의적(p<0.01)으로 높았고, magnesium, calcium에서도 유사한 결과를 보였다(p<0.05). 그러나 G-GTP의 경우 다른 그룹에 비해 그룹 d에서 유의적(p<0.05)으로 가장 낮아졌다. 그러므로 시료채취 시 농가와 실험실의 거리를 고려하여 온도와 저장시간이 혈청 내 사이토카인 및 생화학 농도에 미치는 영향을 최소화 하여 연구 수행을 진행해야 될 것으로 사료된다.
Abstract
Serum plays a role as biomarker in the assessment of health condition or severity of diseases. The separation method, storage temperature and duration affect the levels of cytokines and its biochemical constituents. In this study, blood sampling was performed from Hanwoo cattle through jugular venipuncture. All blood samples were kept at 26±3℃ for 1~2 h and then separated into four conditions (groups a-d). For group (a), the serum was immediately separated (control group) and stored at -70℃; for group (b) the separated serum was kept in ice box for 2 h and then stored at -70℃; for group (c) the collected blood samples were stored in ice box for 2hrs before serum separation; and for group (d); the whole blood was kept at 4℃ for 24hrs before serum separation and then stored at -70℃. All serum samples were stored at -70℃ until analysis. Depending on serum separation time, storage period and temperature, IFN-γ level revealed no significant differences; whereas, TNF-α level was significantly lower (p<0.05) in group (d) than in groups (a-c). Besides, concentration of albumin and total protein were significantly higher (p<0.01) in (d) than that in group (a). The levels of magnesium and calcium also echoed similar results (p<0.05). But G-GTP was significantly lower (p<0.05) in group (d) compared with other groups. Therefore, considering the distance between the farm and the laboratory, it is necessary to minimize the effect of temperature and storage time on serum cytokine and biochemical concentration.
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