痛风宁含药血清对尿酸盐刺激的THP-1炎症反应的时效观察

Time-effect Observation of Serum Containing Tongfengning on Uric Acid Induced Thp-1 Inflammation

目的:观察痛风宁对尿酸盐刺激的人单核细胞THP-1炎症反应的影响,揭示痛风宁发挥最佳抗炎效应的时间。方法:将THP-1随机分为单钠尿酸盐(MSU)造模组和空白对照组,MSU造模组给予MSU混悬液刺激诱导建立细胞炎症模型,空白对照组不予干预。将造模成功的THP-1再随机分为模型组、痛风宁组及阳性对照组,分别用10%空白血清、10%痛风宁含药血清、10%空白血清+Caspase-1抑制剂干预,空白对照组给予10%空白血清干预。于干预后6、12、24、48 h,检测各组THP-1中NALP3、Caspase-1、ASC蛋白、mRNA表达及IL-1β、前列腺素E_(2)(PGE_(2))含量。结果:干预后各时间点,模型组NALP3、Caspase-1、ASC蛋白与mRNA表达及IL-1β、PGE_(2)含量均高于空白对照组(P<0.01或P<0.05);各时间点痛风宁组所有指标水平明显低于模型组(P<0.01或P<0.05);各时间点阳性对照组IL-1β、PGE_(2)含量及Caspase-1蛋白表达均明显低于模型组(P<0.01或P<0.05),NALP3、ASC蛋白及3个mRNA表达差异无统计学意义(均P>0.05);干预后12、24、48 h痛风宁组所有指标水平均低于阳性对照组;痛风宁组各时间点间比较差异有统计学意义(P<0.01或P<0.05);在痛风宁首次干预达24 h时,炎性体相关蛋白及mRNA、炎症介质表达水平达到最低。结论:痛风宁干预尿酸盐刺激的THP-1后,于6 h、12 h、24 h、48 h均表现出显著抗炎效应,在首次干预后24 h抗炎效应达到高峰。

Objective:To observe the effects of Tongfengning on uric acid induced THP-1 inflammation to reveal the optimal time of intervention.Methods:THP-1 was randomly divided into an MSU model establish group and a blank control group.The former received MSU suspension to establish the cell model of inflammation,and the latter received no intervention.After the successful identification of the model,THP-1 in the MSU model establish group was randomly divided into a model group,a Tongfengning group and a positive control group and received 10%blank serum,10%serum containing Tongfengning,10%blank serum+Caspase-1 inhibitor for intervention,respectively.The blank control group received 10%blank serum intervention.At 6,12,24,48 h after the intervention,the expression of NALP3,Caspase-1,ASC protein,mRNA and the content of IL-1βand PGE_(2) in THP-1 in each group were detected.Results:At all point-in-time,compared with the blank control group,the protein and mRNA expression of NALP3,Caspase-1,ASC and the levels of IL-1β,PGE_(2) in the model group were higher(P<0.01 or P<0.05).At all point-in-time,the levels of all indexes in Tongfengning group were lower than the model group(P<0.01 or P<0.05).The levels of IL-1β,PGE_(2) and the expression of Caspase-1 protein in the positive control group at each point-in-time were significantly lower than those in the model group(P<0.01 or P<0.05),and there was no difference in the expression of NALP3,ASC protein and mRNA.Compared with the positive control group,the indexes of Tongfengning group showed significant differences(P s<0.05)at 12,24,48 hours after intervention.There were differences between the expressions of indexes in Tongfengning group at different point-in-time(P<0.01 or P<0.05).When Tongfengning intervened for 24 hours for the first time,the expression levels of inflammasome-related proteins,mRNA and inflammatory factors reached the lowest.Conclusion:Tongfengning intervenes THP-1 stimulated by MSU for 6,12,24 and 48 hours showed significant effect of inhibiting inflammatory response.Especially the anti-inflammatory effect reaches a peak at 24 hours after the first intervention.