木瓜蛋白酶结合电刺激建立兔膝关节创伤性滑膜炎模型的实验研究

Experimental Study on the Establishment of Traumatic Synovitis of Knee Joint Model in Rabbits Using Papain Combined with Electrical Stimulation

目的探讨木瓜蛋白酶联合电刺激建立膝关节创伤性滑膜炎动物模型的可行性。方法将12只新西兰兔随机分为正常组和模型组,每组各6只。从造模开始的第1、4、7天,将0.1 mL的4%木瓜蛋白酶水溶液注射于兔右后肢关节腔内,第8天开始,将模型兔麻醉后仰卧位固定,将电刺激针插入右后肢的股直肌,正极直刺入股直肌中点,负极针直刺入股直肌近端止点上,对应连接BL-420S生物信息采集系统的刺激器,调整参数,在自动串刺激模式下被动牵拉肌肉使膝关节重复被动屈伸,继续造模7 d,对侧肢体只针刺不通电,每天进行1次,每次2 h。造模结束后,对各组采用Lequesne MG评分进行行为学观察;采用游标卡尺测量膝关节肿胀程度、电子体温枪测量兔右膝关节皮肤温度;采用数字触诊仪(Myoton Pro)检测股直肌的肌肉张力、弹性、硬度。采用ELISA法检测血清中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、P物质(SP)的表达水平;HE染色法观察滑膜病理组织情况。结果两组兔行为学Lequesne MG评分、膝关节皮温、周径及血清中IL-1β、TNF-α、SP以及兔右后肢股四头肌肌肉张力、弹性、硬度比较,差异均具有统计学意义(P<0.05);滑膜组织的病理结果显示炎症改变。结论木瓜蛋白酶结合电刺激建立膝关节创伤性滑膜炎模型成功。

Objective:To investigate the method of establishing an animal model of traumatic synovitis of knee joint(TSKJ)by papain combined with electrical stimulation.Methods:12 New Zealand rabbits were randomly divided into a normal group and a model group,with 6 rabbits in each group.Starting from day 1 of the modeling experiment,0.1 mL of a 4%papain aqueous solution was injected into the right hindlimb joint cavity of the rabbits in the model group on days 1,4,and 7.From day 8,the rabbits in the model group were placed in a supine position under anesthesia,and electrical stimulation needles were inserted into the rectus femoris muscle of the right hindlimb.The positive electrode was directly inserted into the midpoint of the rectus femoris,and the negative electrode needle was inserted near the proximal insertion point of the muscle,connected to the stimulator of the BL-420s biological information acquisition system.Parameters were adjusted,and under the automatic train stimulation mode,the muscles were passively stretched to repeatedly flex and extend the knee joint for 7 more days to continue the modeling process.The contralateral limb was only needled without electrical stimulation,with the procedure performed once daily for 2 h each time.After the modeling process,behavioral observations were first conducted on each group using the Lequesne MG score.The degree of knee joint swelling was measured using a vernier caliper,and the skin temperature of the right knee joint of the rabbits was measured using an infrared thermometer.The muscle tension,elasticity,and hardness of the rectus femoris were detected using a digital palpation device(Myoton Pro).The serum levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and substance P(SP)were detected using the ELISA method.The histopathological condition of the synovial tissue was observed using the hematoxylin-eosin(HE)staining method.Results:Comparisons of the Lequesne MG scores,skin temperature around the knee joint,circumference,and serum levels of IL-1β,TNF-α,SP,as well as the muscle tension,elasticity,and hardness of the quadriceps muscle in the right hindlimb of the rabbits between the two groups showed statistically significant differences(P<0.05).The histopathological results of the synovial tissue indicated inflammatory changes.Conclusion:The successful establishment of a TSKJ model in rabbit was achieved using a combination of papain and electrical stimulation。