基于NALP3炎性体的痛风宁对急性痛风性关节炎大鼠抗炎作用量效和时效研究

Study on Tongfengning on Dosage Effect and Time Effect of Anti-inflammatory in Model Rats with Acute Gouty Arthritis Based on NALP3 Inflammasome

目的观察痛风宁对急性痛风性关节炎(AGA)大鼠NALP3炎性体及相关炎症因子的影响,探讨其抗炎作用的量效与时效关系。方法将240只SD大鼠随机分为造模组200只和正常组40只。右后踝关节腔内注射尿酸盐混悬液制作大鼠AGA模型,成模大鼠随机分为模型组、秋水仙碱组及痛风宁低、中、高剂量组,每组40只。正常组和模型组大鼠给予等量生理盐水灌胃,其余组大鼠给予相应药物灌胃干预。首次干预后6、12、24、48 h各组随机取10只大鼠处死,收集右后踝关节液与滑膜组织,采用ELISA、Western blot和RT-qPCR分别检测白细胞介素-1β(IL-1β)、前列腺素E2(PGE2)含量及NALP3、ASC、Caspase-1蛋白和mRNA表达。结果与正常组比较,模型组大鼠各时间点IL-1β、PGE2含量及NALP3、ASC、Caspase-1蛋白和mRNA表达明显升高(P<0.01);与模型组比较,痛风宁中、高剂量组及秋水仙碱组大鼠各时间点IL-1β、PGE2含量及NALP3、ASC、Caspase-1蛋白和mRNA表达均明显降低(P<0.01,P<0.05);与秋水仙碱组比较,痛风宁中、高剂量组大鼠各时间点IL-1β、PGE2含量及NALP3、ASC、Caspase-1蛋白和mRNA表达无明显差异(P>0.05),痛风宁低剂量组大鼠明显升高(P<0.01,P<0.05)。结论中剂量痛风宁在下调模型大鼠NALP3、ASC、Caspase-1蛋白和mRNA表达及降低IL-1β、PGE2含量方面作用相对较佳,且首次干预后48 h抗炎作用相对明显。

Objective To observe the effects of Tongfengning on NALP3 inflammasome and related inflammatory factors in acute gouty arthritis(AGA)rats;To explore the dosage effect and time effect of its anti-inflammatory effect.Methods Totally 240 SD rats were randomly divided into model rats(n=200)and normal rats(the normal group,n=40).The AGA model was made by intraperitoneal injection of urate suspension into the right posterior ankle joint of the rats.Model rats were randomly divided into model group,colchicine group and Tongfengning low-,mediumand high-dosage groups,with 40 rats in each group.Normal group and model group were given the same amount of normal saline for gavage,and other groups were given the corresponding medicine for gavage intervention.At 6 h,12 h,24 h,and 48 h after the first intervention,10 rats in each group were sacrificed at each time point,and the right posterior ankle joint fluid and synovial tissue were collected.ELISA,Western blot,and RT-qPCR were used to detect the contents of interleukin-1β(IL-1β)and prostaglandin E2(PGE2),and expressions of NALP3,ASC,Caspase-1 protein and mRNA.Results Compared with the normal group,the contents of IL-1βand PGE2,and the expressions of NALP3,ASC,Caspase-1 protein and mRNA increased in the model group at each time point(P<0.01);Compared with the model group,the contents of IL-1βand PGE2,and the expressions of NALP3,ASC,Caspase-1 protein and mRNA significantly decreased in Tongfengning medium-and high-dosage groups at each time point(P<0.01,P<0.01);Compared with the colchicine group,the contents of IL-1βand PGE2,and the expressions of NALP3,ASC,Caspase-1 protein and mRNA were not significantly different in the Tongfengning medium-and high-dosage groups at each time point(P>0.05),while they increased significantly in Tongfengning low-dosage group(P<0.01,P<0.05).Conclusion The medium dosage of Tongfengning is relatively better in down-regulating the expressions of NALP3,ASC,Caspase-1 protein and mRNA and decreasing the contents of IL-1βand PGE2,and its anti-inflammatory effects were relatively obvious 48 h after the first intervention.