铁包金按摩膏联合刮法对膝骨关节炎模型兔关节软骨IL-1β、TNF-α及A20蛋白表达的影响

Effects of Tiebaojin Anmo Ointment(铁包金按摩膏)Combined with Scraping Method on the Expression of IL-1β,TNF-αand A20 Protein in Articular Cartilage of Knee Osteoarthritis Model Rabbits

目的:探索铁包金按摩膏联合刮法对膝骨关节炎模型兔膝关节软骨组织中TNF-α、IL-1β及锌指蛋白A20表达的影响。方法:采用随机数字表法从40只实验兔中选出8只(雌雄各半)作为正常组,仅常规饲养,不做处理。其余32只实验兔制作兔KOA模型。造模成功后再将32只实验兔采用随机数字表法分成模型组、刮法组、涂擦组、药刮组,每组8只(雌雄各半)。造模1周后,各组实验兔给予相应干预,连续4周。末次给药24 h后取关节软骨,制作病理切片观察软骨病理改变;采用ELISA法检测软骨组织中TNF-α、IL-1β水平;采用Western blotting法检测软骨组织中锌指蛋白A20的表达。与正常组比较,模型组、刮法组、涂擦组及药刮组兔膝关节Lequesne MG评分、软骨组织Mankin评分、软骨组织中TNF-α和IL-1β水平均明显升高(P<0.05),而软骨组织中A20表达均明显下调(P<0.05);与模型组比较,刮法组、涂擦组及药刮组兔膝关节Lequesne MG评分、软骨组织Mankin评分、软骨组织中TNF-α和IL-1β水平均明显降低(P<0.05),而软骨组织中A20表达均明显上调(P<0.05);刮法组兔膝关节各检测指标与涂擦组比较,差异无统计学意义(P>0.05);药刮组兔膝关节各检测指标均优于刮法组、涂擦组(P<0.05)。结论:铁包金按摩膏联合刮法对KOA模型兔膝骨关节炎具有一定治疗作用,其作用机制可能与抑制TNF-α、IL-1β表达和促进A20表达有关。

Objective:To explore the effect of Tiebaojin Anmo Ointment combined with scraping method on the expression of TNF-α,IL-1β and zinc finger protein A20 in knee articular cartilage tissue of knee osteoarthritis model rabbits.Methods:8 rabbits(half male and half female)were selected from 40 experimental rabbits as the normal group by random number table method.They were only fed conventionally without treatment.The other 32 experimental rabbits made rabbit KOA model.After successful modeling,32 experimental rabbits were randomly divided into model group,scraping group,rubbing group and medicine scraping group,with 8 rabbits in each group(half male and half male).One week after modeling,the experimental rabbits in each group were given corresponding interventions for 4 consecutive weeks.24 h after the last administration,articular cartilage was collected,and pathological sections were made to observe the pathological changes of cartilage.The levels of TNF-α and IL-1β in cartilage tissue were detected by ELISA.Western blotting method was used to detect the expression of zinc finger protein A20 in cartilage tissue.Results:Compared with the normal group,the Lequesne MG score,Mankin score of cartilage tissue,and the levels of TNF-α and IL-1β in the cartilage tissue of the model group,scraping group,rubbing group and medicine scraping group were significantly increased(P<0.05),while the expression of A20 in cartilage tissue was significantly down-regulated(P<0.05).Compared with the model group,the Lequesne MG score,the Mankin score of cartilage tissue,and the levels of TNF-α and IL-1β in cartilage tissue in the scraping group,rubbing group and medicine scraping group were significantly decreased(P<0.05),while the expression of A20 in cartilage tissue was significantly up-regulated(P<0.05).There was no significant difference between the scraping group and the rubbing group(P>0.05).The indexes of rabbit knee joint in medicine scraping group were better than those in scraping group and rubbing group(P<0.05).Conclusion:Tiebaojin Anmo Ointment combined with scraping method has a certain therapeutic effect on KOA model rabbit knee osteoarthritis,and its mechanism may be related to inhibiting the expression of TNF-α and IL-1β and promoting the expression of A20.