青藤碱对胶原诱导性关节炎大鼠治疗作用

Therapeutic effect of sinnomenine on collagen-induced arthritis in rats

目的探讨青藤碱对Ⅱ型胶原蛋白诱导性关节炎大鼠的治疗作用及其机制。方法 6周龄SPF级雌性Wistar大鼠40只,随机分为对照组、模型组、青藤碱组和甲氨蝶呤组各10只。对照组大鼠不予处理,模型组、青藤碱组和甲氨蝶呤组制备关节炎大鼠模型后,分别采用生理盐水10mL/(kg·d)、青藤碱100mg/(kg·d)和甲氨蝶呤40mg/(kg·d)连续灌胃28d。观察青藤碱对关节炎大鼠足容积和关节肿胀评分的影响;采用X线影像学分析大鼠关节骨侵蚀和关节间隙变化;采用ELISA法检测血清肿瘤坏死因子-α和白细胞介素-1β水平。结果模型组第1、2、3、4周足容积[(0.94±0.17)、(1.12±0.41)、(1.46±0.64)、(1.28±0.75)mL]明显高于对照组[(0.51±0.13)、(0.51±0.08)、(0.53±0.12)、(0.54±0.16)mL]、青藤碱组[(0.62±0.10)、(0.68±0.13)、(0.73±0.19)、(0.87±0.23)mL]和甲氨蝶呤组[(0.53±0.11)、(0.60±0.10)、(0.62±0.14)、(0.66±0.17)mL](P<0.05),青藤碱组与甲氨蝶呤组各时间点比较差异均无统计学意义(P>0.05);模型组第1、2、3、4周时关节肿胀评分[(2.1±0.3)、(7.9±0.5)、(11.7±1.2)、(10.4±0.9)分]均明显高于对照组(均为0分)和甲氨蝶呤组[(1.5±0.2)、(4.6±0.5)、(5.1±0.6)、(4.9±0.5)分],第3、4周关节肿胀评分高于青藤碱组[(7.2±0.8)、(6.1±0.7)分](P<0.05);第4周模型组关节间隙狭窄评分[(143.18±39.50)分]和总评分[(321.39±104.80)分]高于对照组(均为0分)、青藤碱组[(78.67±25.38)、(191.15±67.75)分]和甲氨蝶呤组[(61.72±21.24)、(158.26±51.40)分](P<0.05),模型组骨侵蚀评分[(178.21±65.49)分]明显高于对照组(0分)和甲氨喋呤组[(96.54±30.16)分](P<0.05);模型组大鼠第30天血清肿瘤坏死因子-α[(49.72±6.58)μg/L]、白细胞介素-1β[(46.94±5.07)μg/L]水平均明显高于对照组[(14.96±2.37)、(11.85±2.46)μg/L]、青藤碱组[(24.10±4.12)、(21.02±3.88)μg/L]和甲氨蝶呤组[(20.56±4.63)、(19.73±4.41)μg/L](P<0.05)。结论青藤碱对胶原诱导性关节炎大鼠有明显治疗作用,可能与其抑制血促炎细胞因子升高有关。

Objective To investigate the therapeutic effect of sinnomenine on typeⅡcollagen-induced arthritis（CIA）in rats and its related anti-inflammatory mechanisms.Methods Forty 6-week-old female Wistar rats（SPF grade）were randomly divided into control group,model group,sinnomenine group and methotrexate group,with 10 rates in each group.Control group received no treatment.Model group,sinnomenine group and methotrexate group received lavage by normal saline（10mL/（kg·d））,sinnomenine（100mg/（kg·d））and methotrexate（40mg/（kg·d））respectively for 28 days after the establishment of CIA rat models.The influences of sinnomenine on paw volume and arthritis index were observed.The radiological changes of joint damage including rosion and joint space narrowing were analyzed by X-ray iconography.Serum pro-inflammatory cytokines levels as tumor necrosis factor-alpha（TNF-α）and interkeukin-1beta（IL-1β）were measured by ELISA.Results The paw volumes in 1,2,3and 4weeks were significantly higher in model group（（0.94±0.17）,（1.12±0.41）,（1.46±0.64）,（1.28±0.75）mL）than those in control group（（0.51±0.13）,（0.51±0.08）,（0.53±0.12）,（0.54±0.16）mL）,sinnomenine group（（0.62±0.10）,（0.68±0.13）,（0.73±0.19）,（0.87±0.23）mL）and methotrexate group（（0.53±0.11）,（0.60±0.10）,（0.62±0.14）,（0.66±0.17）mL）（p〈0.05）,and there were no significant differences between sinnomenine group and methotrexate group（p〈0.05）.The joint swelling indexes were significantly higher in 1,2,3and 4weeks in model group（2.1±0.3,7.9±0.5,11.7±1.2,10.4±0.9）than those in control group（0）and methotrexate group（1.5±0.2,4.6±0.5,5.1±0.6,4.9±0.50）,and were higher in 3and 4weeks in model group than those in sinnomenine group（7.2±0.8,6.1±0.7）（p〈0.05）.The joint space narrowing scores and total scores were higher in model group（143.18±39.50,321.39±104.80）in 4weeks than those in control group（0）,sinnomenine group（78.67±25.38,191.15±67.75）and methotrexate group（61.72±21.24,158.26±51.40）（p〈0.05）.The bone erosion score was significantly higher in model group（178.21±65.49）than that in control group（0）and methotrexate group（96.54±30.16）（p〈0.05）,The serum concentrations of TNF-αand IL-1βwere significantly higher in model group（（49.72±6.58）,（46.94±5.07）μg/L）on the 30 th day than those in control group（（14.96±2.37）,（11.85±2.46）μg/L）,sinnomenine group（（24.10±4.12）,（21.02±3.88）μg/L）and methotrexate group（（20.56±4.63）,（19.73±4.41）μg/L）（p〈0.05）.Conclusion Sinomenine has an obvious antiinflammatory action on CIA rats,which is associated with its inhibitory effects on the production of pro-inflammatory cytokines.