胶原诱导的关节炎模型 (CIA)
实验方案
1. 牛II型胶原溶解在0.05mol/ml的乙酸中,浓度为2mg/ml,搅拌4°C过夜。
2. 注射前,II胶原溶液与同体积的IFA在冰浴中*乳化。
3. *天,每只大鼠尾根部皮内注射0.1ml CII/IFA乳化剂(浓度为1mg/ml)。
4. 第7天,每只大鼠以同样的方式再次注射。
5. 注射后,大鼠关节炎指数根据肉眼评分系统每周两次进行评价。
0-正常
1-一个足爪肿胀和/或发红
2-2个相关关节肿胀和/或发红
3->2个相关关节肿胀和/或发红
4-整个足爪严重炎症
每只大小鼠关节炎指数为每只动物4足爪得分之和。
6. 肿胀严重性根据器官充满度测定法通过气道容积的改变来测定, 一周两次。
Brief protocol of Collagen induced arthritis model (CIA)
1. Bovine CII is dissolved in 0.05mol/L acetic acid at a concentration of 2 mg/ml by stirring overnight at 4°C.
2. Before injection, CII is emulsified with an equal volume of IFA in an ice-cold water bath.
3. On day1, each rat is injected intradermally at the base of the tail with 0.1ml CII/IFA the emulsion (1mg/ml).
4. On day 21, a second injection of CII in IFA will be administered using the same amount.
5. After the second injection,rats will be evaluated daily for arthritis using a macroscopic scoring system of 0–4.
0 no signs of arthritis
1 swelling and/or redness of the paw or 1 digit
2=2 joint involved
3=>2 joint involved
4severe arthritis of the entire paw and digits
An arthritis index for each rat is calculated by adding the scores for each of the 4 paws.
6. Swelling severity is also determined by quantitating the change in the paw volume, as measured by plethysmometry.
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