清热除湿通络法对大鼠痛风性关节炎模型中NALP3炎性体表达的影响

Effects of clearing heat and removing dampness and dredging collateral method on expression of NALP3 inflammasome in model of gouty arthritis rats

目的:观察清热除湿通络法对大鼠痛风性关节炎模型中NALP3、Caspase-1、Pycard m RNA及NALP3、Caspase-1蛋白表达的影响。方法:将经典痛风性关节炎模型大鼠造模后采用随机数表法分为空白组、模型组、虎杖组及其各拆方组、吲哚美辛组和秋水仙碱组。造模12h后取材,随后运用RT-PCR技术检测大鼠气囊灌洗液中NALP3、Caspase-1、Pycard m RNA变化;Western blot法检测大鼠气囊灌洗液中NALP3、Caspase-1蛋白的变化。结果:以模型组为对照,虎杖组NALP3 m RNA相对表达下调,而其余组均上调;虎杖组Caspase-1 m RNA上调幅度小于其余组(除清热方组);虎杖组Pycard m RNA表达上调幅度低于其余组。虎杖组NALP3蛋白表达水平较模型组及其各拆方组、秋水仙碱组低(P<0.01,P<0.05);虎杖组Caspase-1蛋白表达量低于除湿方组、模型组(P<0.05,P<0.01)。结论:清热除湿通络法通过在蛋白质翻译层面下调NALP3、Caspase-1蛋白,干扰了NALP3炎性复合体的装配或激活,进而发挥了拮抗痛风炎性反应的作用。

Objective： To observe the effects of clearing heat and removing dampness and dredging collateral method on the expression of NALP3, Caspase- 1, Pycard mRNA and NALP3, Caspase-1 protein in the model of gouty arthritis rats. Methods： Model of gouty arthritis rats were randomly divided into the blank group, the model group, the indomethacin group, the colchicine group, the Huzhang Tongfeng Granule group and its disassembled prescription group. The expressions of NALP3, Caspase-1, and Pycard mRNA in airbag lavage fluid were analyzed by RT-PCR. NALP3 and Caspase-1 protein were analyzed by Western blot. Results： Compared with the model group, the expression of NALP3 mRNA in the Huzhang Tongfeng Granule group was down-regulated, while the other groups were up-regulated; the up-regulation of Caspase-1 mRNA in Huzhang Tongfeng Granule group was less than that of the other groups, except for the Clearing Heat Formula group; the up-regulation of the expression of Pycard mRNA in the Huzhang Tongfeng group was lower than that of the other groups. The expression level of NALP3 protein in Huzhang Tongfeng Granule group of was lower than that of the model group, the disassembled prescription group and the colchicine group （P〈0.01, P〈0.05）. The expression of Caspase-1 protein in Huzhang Tongfeng Granule group was lower than that of the Removing Dampness Formula group and the model group （P〈0.05, P〈0.01）. Conclusion： Clearing heat and removing dampness and dredging collateral method plays a role in antagonizing gouty inflammation by downregulating NALP3 and Caspase-1 protein at the protein translation level, interfering with the assembly or activation of NALP3 inflammatory complex.