健脾理气方通过下调NLRP3炎症小体的活化防治内毒素性急性肝损伤大鼠

目的:在网络药理学研究基础上,基于NLRP3炎性小体活化通路探讨健脾理气方对大鼠急性肝损伤(ALI)的防治作用。方法:(1)利用网络药理学技术预测分析健脾理气方通过抗炎和免疫调节治疗ALI的潜在靶点。(2)将雄性Sprague-Dawley大鼠随机分为正常对照组、模型对照组以及健脾理气方6. 05、12. 1、24. 2 g/kg组,除正常对照组大鼠按体重一次性腹腔注射生理盐水外,其余各组大鼠给予腹腔注射600 mg/kg D-氨基半乳糖(D-GalN)诱导ALI模型。造模后健脾理气方6. 05、12. 1、24. 2 g/kg组立即分别灌胃相应体积药物,每日两次,给药间隔为12 h,连续给药2 d。末次给药后立即取材,采用HE染色观察肝脏组织病理形态学的改变;生物化学法检测大鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和γ-谷氨酰转肽酶(GGT)及肝脏组织中的Caspase-1活性,ELISA法检测血清炎症因子IL-1β、IL-18和NF-κB含量; Western Blot法检测肝脏组织中衔接分子(ASC)和炎性小体(NLRP3)蛋白表达水平的变化。结果:(1)网络药理学预测结...

Jianpi Liqi Decoction Alleviates Acute Liver Injury By Down Regulating NLRP3 Inflammasome Activation in D-GalN Induced Rats

Objective: Based on network pharmacology research,the prevention and treatment effect of Jianpi Liqi decoction on acute liver injury(ALI) in rats were explored through NLRP3 inflammatory activation pathway. Methods:( 1) To prodict potential targets of Jianpi Liqi decoction in the treatment of acute liver injury( ALI) through anti-inflammatory and immunomodulation by network pharmacology technology.( 2)Male Sprague-Dawley rats were randomly divided into the control group,the model group and Jianpi Liqi decoction groups( 6. 05、12. 1、24. 2 g/kg). In the control group,rats were intraperitoneally injected saline water,while rats in other groups were intraperitoneal injected 600 mg/kg D-Galactose( D-Gal N),to establish the ALI model. After modeling,all Jianpi Liqi decoction groups( 6. 05、12. 1、24. 2 g/kg) were given corresponding doses,twice a day( 12 h between administration),for 2 days. After the last administration,serum and liver samples were collected. Liver histological morphology changes were observed by HE staining. Activities of glutamate transaminase( ALT),glutamate transaminase( AST) and gamma-glutamine transpeptidease( GGT) in rat serum and activity of caspase-1 in liver tissues were determined by biochemical method. The contents of serum inflammatory factors IL-1 beta,IL-18 and NF-κB were detected by ELISA. The levels of ASC andNLRP3 proteins in liver tissues were detected by Western Blot. Results:( 1) Network pharmacology results indicated that Jianpi Liqi decoction treated ALI through the signaling pathways such as NLRP3 inflammatory.( 2) Compared with the control group,the liver cell damage index was increased in the model group,the contents of ALT,AST,IL-1β NF-κB in serum were increased,the activity of caspase-1 enzyme was increased,the expression levels of NLRP3 and ASC protein and in liver tissues were significantly up regulated( P < 0. 05 or P < 0. 01).Compared with the model group,the damage index of liver cells in ALI rats was significantly decreased in 12. 1 and 24. 2 g/kg Jianpi Liqi decoction groups. The contents of ALT,AST,GGT and NF-κB in serum and the expressions of NLRP3 and ASC in liver tissues were significantly decreased in 12. 1 g/kg group( P < 0. 05 or P < 0. 01). The contents of GGT,IL-18 and NF-κB in serum,the expressions of NLRP3 and ASC in liver tissues and the activity of caspase-1 were also significantly decreased in 24. 2 g/kg group( P < 0. 05 or P < 0. 01).Conclusion: Jianpi Liqi decoction at the doses of 12. 1 and 24. 2 g/kg has a certain protective effect on D-Gal N induced acute liver injury,while 12. 1 g/kg Jianpi Liqi decoction has the best effect. Its mechanisms may be down regulating the activation of NLRP3 inflammatory,which agrees with the network pharmacology prediction results.