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目的观察蜂毒素联合杜仲黄酮外用对于胶原诱导型(CIA)类风湿关节炎小鼠的影响,并观察杜仲黄酮能否有效抑制蜂毒素的溶血作用。方法 1动物实验:采用CIA法诱导40只类风湿关节炎的小鼠,将其随机分为PBS组、蜂毒素组、杜仲黄酮组和蜂毒素联合杜仲黄酮混合给药组,同时取5只未经过任何处理的小鼠组成空白对照组。蜂毒素组每日外用蜂毒素5μl/爪(含药浓度1μg/μl),杜仲黄酮组每日外用杜仲黄酮5μl/爪(含药浓度1μg/μl),混合给药组给药剂量相同(为蜂毒素与杜仲黄酮等比例混合制成,含药浓度为1μg/μl),PBS组每日给等量的PBS,空白对照组不给予任何治疗,连续给药15天观察小鼠的相关变化。2溶血实验:2%红细胞悬浊液中分别加入100μl的蜂毒素溶液,杜仲黄酮溶液,蜂毒素联合杜仲黄酮1∶1溶液,蜂毒素联合杜仲黄酮2∶1溶液,蜂毒素联合杜仲黄酮1∶2溶液,37℃水浴加热后过夜,肉眼及紫外分光光度计检测OD值。结果蜂毒素组、杜仲黄酮组和混合给药组均对类风湿关节炎CIA小鼠在大体观察、足垫厚度、足踝宽度、TNF-α、IL-1β、IL-2、GM-CSF及影像学方面与PBS组相比,均有好转,差异有统计学意义;蜂毒素联合杜仲黄酮组治疗效果好于两种药物单独外用;蜂毒素联合杜仲黄酮1∶1,1∶2溶液不产生红细胞溶血现象,蜂毒素组、蜂毒素联合杜仲黄酮2∶1溶液会使红细胞悬浊液产生溶血现象。结论蜂毒素、杜仲黄酮、蜂毒素联合杜仲黄酮外用对于类风湿关节炎CIA小鼠有治疗效果,其中联合用药效果更好;杜仲黄酮可以有效的抑制蜂毒素的溶血现象。
Objective To observe the effect of melittin and Eucommia ulmoides flavonoids on the collagen induced type (CIA) rheumatoid arthritis in mice and observe whether the Eucommia flavone can effectively inhibit the melittin hemolytic effect. Method 1 Animal experiments: 40 mice with rheumatoid arthritis induced by CIA were randomly divided into PBS group, melittin group, Eucommia flavone group and melittin plus Eucommia ulmoides flavonoids mixed treatment group, taking 5 Mice subjected to any treatment formed a blank control group. Melittin group daily melittin 5μl / claw (drug concentration 1μg / μl), Eucommia flavone group daily external Eucommia flavonoids 5μl / claw (drug concentration 1μg / μl), mixed drug group administered the same dose Melittin and Eucommia flavonoids were mixed in equal proportion, containing drug concentration of 1μg / μl). PBS was given to the same amount of PBS daily, while the blank control group was not given any treatment. The rats were continuously treated for 15 days to observe the changes. 2 hemolysis experiments: 2% erythrocyte suspension were added melittin solution 100μl, Eucommia flavonoids, melittin and Eucommia flavonoids 1: 1 solution, melittin and Eucommia flavonoids 2: 1 solution, melittin and Eucommia flavone 1: 2 solution, 37 ℃ water bath overnight after heating, naked eye and UV spectrophotometer OD value. Results The melittin group, the Eucommia ulmoides flavonoids group and the mixed drug group were all effective in the treatment of rheumatoid arthritis CIA mice in general observation, footpad thickness, ankle width, TNF-α, IL-1β, IL-2, GM- Compared with the PBS group, imaging improved, the difference was statistically significant; melittin combined with Eucommia flavone group than the two drugs alone topical effect; melittin with Eucommia flavone 1: 1, 1: 2 solution does not produce Red blood cell hemolysis, melittin group, melittin and Eucommia flavone 2: 1 solution will hemolysis phenomenon of erythrocyte suspension. Conclusion Melittin, Eucommia flavone, melittin and Eucommia ulmoides flavonoids have therapeutic effects on rheumatoid arthritis CIA mice, and the combined effect is better. Eucommia flavonoids can effectively inhibit the melittin hemolytic phenomenon.