解毒化瘀颗粒对急性肝衰竭大鼠NLRP3炎症体调控作用的实验研究

Experimental Study on the Regulatory Effect of Jiedu Huayu Granule on NLRP3 Inflammatory Body of Rats with Acute Liver Failure

目的:观察解毒化瘀颗粒对急性肝衰竭大鼠NLRP3炎症体的调控作用,探索解毒化瘀颗粒治疗急性肝衰竭的作用机制。方法:SPF级SD大鼠36只,随机分为解毒化瘀颗粒组12只、模型组12只和空白对照组12只。解毒化瘀颗粒组和模型组大鼠给予腹腔注射D-GalN 600mg/kg和LPS 20μg/kg制备急性肝衰竭大鼠模型,空白对照组给予腹腔注射等体积生理盐水。造模前1周开始灌胃给药,解毒化瘀颗粒组给予解毒化瘀颗粒灌胃,模型组与空白对照组给予等体积生理盐水灌胃。于造模成功后第3天取材,采集血液标本及大鼠肝组织,HE染色观察,运用Western Blot、qPCR检测肝组织中NLRP3、Caspase-1、ASC蛋白和mRNA表达情况;运用ELISA检测血清IL-1β、IL-18表达情况。结果:解毒化瘀颗粒组大鼠血清IL-1B、IL-18水平低于模型组,差异具有统计学意义(P<0.05)。与模型组对比,解毒化瘀颗粒组大鼠肝组织中NLRP3、Caspase-1、ASC蛋白及mRNA表达量降低,差异具有统计学意义(P<0.05)。结论:解毒化瘀颗粒能够下调NLRP3炎症体NLRP3、Caspase-1、ASC、IL-1β和IL-18等相关因子的表达,对急性肝衰竭具有一定的治疗作用。

Objective:To observe the regulatory effect of Jiedu Huayu Granule on NLRP3 inflammatory body in rats with acute liver failure,and to explore the mechanism of Jiedu Huayu Granule in treating acute liver failure.Methods:36 SPF SD rats were randomly divided into Jiedu Huayu Granule group(n=12),model group(n=12)and blank control group(n=12).Jiedu Huayu Granule group and model group were given intraperitoneal injection of D-GalN 600mg/kg and LPS 20μg/kg to prepare rat model of acute liver failure,while blank control group was given intraperitoneal injection of equal volume of normal saline.1 week before modeling,the Jiedu Huayu Granule group was given gavage of Jiedu Huayu Granule,and the model group and blank control group were given equal volume normal saline gavage.On the 3rd day after the successful modeling,blood samples and rat liver tissues were collected and observed by HE staining.NLRP3,Caspase-1,ASC protein and mRNA expression in liver tissues were detected by Western Blot and qPCR.The expression of IL-1βand IL-18 in serum was detected by ELISA.Results:The serum levels of IL-1βand IL-18 in Jiedu Huayu Granule group were lower than those in model group,and the difference was statistically significant(P<0.05).Compared with model group,the expressions of NLRP3,Caspase-1 and ASC protein and mRNA in liver tissues of rats in Jiedu Huayu Granule group were decreased,with statistical significance(P<0.05).Conclusion:Jiedu Huayu Granule can down-regulate the expression of NLRP3,Caspase-1,ASC,IL-1β,IL-18 and other related factors in NLRP3 inflammatory body,which has certain therapeutic effect on acute liver failure.