二甲双胍对Ⅱ型胶原诱导类风湿性关节炎大鼠模型的抗炎及关节保护作用的研究

Effect of metformin in vivo on the inhibition of inflammation and joint protection of the collagen-induced arthritis rat model

目的 研究二甲双胍对Ⅱ型胶原诱导类风湿性关节炎大鼠模型的抗炎及关节保护的作用。方法 4℃下,将Ⅱ型胶原溶于醋酸中,制成混合液;再与完全弗氏佐剂以1∶1相配比制成乳剂。于大鼠尾部、肛周皮肤多点皮内注射0.3 mL乳剂;实验第7天重复操作,加强诱导1次.造模成功后,于实验第7天将大鼠随机分为5组,为高剂量二甲双胍组、低剂量二甲双胍组、甲氨蝶呤组、诱导组、正常对照组,每组10只,分别给予高剂量二甲双胍（100 mg·kg^-1·d^-1）,低剂量二甲双胍（50 mg· kg^-1·d^-1）,甲氨蝶呤（2.7 mg·kg^-1·w^-1）灌胃干预治疗,诱导组和正常对照组不给予药物,每隔2d对各组大鼠进行关节炎评分及距骨宽度测量;于第28天处死取材,再进行大鼠踝关节X线片,大鼠血清中炎症因子水平及抗炎因子水平等检测。结果 连续观察大鼠后足关节炎评分及距骨宽度测量示：高剂量二甲双胍组关节炎评分及距骨宽度较诱导组显著降低（P＜0.05）,其余两用药组较诱导组也有一定降低关节炎评分及距骨宽度的作用（P＜0.05）;X线片示：高剂量二甲双胍组可显著保留关节结构并使其免受侵蚀,而剂量低二甲双胍组和甲氨蝶呤组效果比高剂量二甲双胍组较弱,关节结构破坏程度较轻;ELISA结果示：高剂量二甲双胍组可显著降低大鼠血清中炎症因子TNF-α、IL-1β、IL-6水平（P＜0.05）,升高抗炎因子IL-10水平（P＜0.05）;低剂量二甲双胍组及甲氨蝶呤组大鼠血清中炎症因子TNF-α、IL-1β、IL-6水平较诱导组有所降低（P＜0.05）,抗炎因子IL-10水平也有升高（P＜0.05）。结论二甲双胍对于Ⅱ型胶原诱导类风湿性关节炎大鼠模型有显著的抗炎及关节保护的作用。

Objective To verify the effect of metformin in vivo on the inhibition of inflammation and joint protection of the collagen-induced arthritis （CIA） rat model.Methods Under 4 ℃,dissolved type]Ⅱ collagen into acetic acid.In order to made the mixture into emulsion,blended it with complete freund＇s adjuvant to 1∶ 1.Gave several spots intraperitoneal injection of 0.3 mL emulsion in the rat tail and the skin around anus.Repeat the experi-mental operation on the 7th day to strengthen the induction.After the success of the induction,gave CIA rat models high doses of metformin（100 mg·kg^-1 · d^-1）,low doses of metformin（50 mg · kg^-1 · d^-1） and methotrexate （2.7 mg · kg-1 · w^-1） for treatment by gavage on the 7th day.Evaluated the arthritis score of each group of rats and measured the width of talus every two days.Executed rats and drew materials on the 28th day,then shot X-ray films of the ankle of rat and detected the levels of inflammatory and anti-inflammatory factors in rat serum by Elisa.Results After continuous observation rat foot arthritis scoring and measurement from talus width：arthritis scoring of high doses of metformin group was obviously lower than other induced groups （P 〈 0.05）,talus width is also much smaller（P 〈 0.05） ; the rest of the treatment groups also has a certain effect on reducing arthritis score and the width of talus compared to the induced group （P 〈 0.05）.X-ray radiography show：high doses of metformin group can significantly save joint structure and make joint clearance clear; the treatment effect of the rest treatment groups are not better than high dose group,also has severe soft tissue swelling,bone absorption,joint disintegration and severe damage.Elisa show：High doses of metformin can lower the level of inflammatory factors-TNF-α,IL1 β,IL-6 （P 〈 0.05）,also can rise the level of anti-inflammatory factor-IL-10 obviously（P 〈 0.05）.Low doses of metformin and MTX have the similar effect （P 〈 0.05）,but not more significant than high doses of metformin.Conclusions Metformin in vivo can significantly Inhibit the inflammation and protect the joint structure of the collagen-induced arthritis （CIA） rat model.