荔枝核总黄酮抑制NLRP3炎症小体减轻小鼠急性肝损伤的作用

Effect of Lychee Seed Flavonoids on Inhibiting NLRP3 Inflammasome and Alleviating Acute Liver Injury in Mice

目的 探讨荔枝核总黄酮对急性肝损伤的保护作用,及抑制NOD样受体家族3(NOD-like receptor family 3,NLRP3)炎症小体活化相关的机制。方法 将昆明小鼠随机分为空白组,模型组,联苯双酯组,MCC950组,荔枝核总黄酮高、低剂量组。除空白组外,其余以腹腔注射0.2%CCl4-花生油溶液诱发小鼠急性肝损伤,观察各组小鼠肝功能,肝脏组织形态,肝组织活性氧族(ROS)水平,并以Western blot法检测肝组织中NLRP3炎症小体相关蛋白的表达变化。结果 与空白组相比,模型组小鼠血清中丙氨酸氨基转氨酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆汁酸(TBA)含量,肝组织中ROS水平均显著升高(P<0.05),肝组织中硫氧还蛋白结合蛋白(TXNIP)、NLRP3、半胱氨酸蛋白酶-1前体(pro-caspase-1)、白细胞介素-1β前体(pro-IL-1β)、白细胞介素-1β(IL-1β)蛋白表达显著增加(P<0.05),肝脏组织损伤程度严重;与模型组相比,荔枝核总黄酮剂量组显著降低CCl4所致急性肝损伤的小鼠血清中ALT、AST、TBA水平(P<0.05),降低肝组织中ROS水平(P<0.05),并减轻肝组织结构损伤,降低HE染色的形态学评分(P<0.05)。荔枝核总黄酮高剂量与MCC950组均显著下调了NLRP3炎症小体相关蛋白TXNIP、NLRP3、pro-caspase-1、pro-IL-1β、IL-1β表达(P<0.05)。结论 荔枝核总黄酮减轻了CCl4引发的小鼠急性肝损伤,其机制可能与抑制NLRP3炎症小体活化参与的炎症反应有关。

Objective To investigate the protective effect of lychee seed flavonoids on acute liver injury and the mechanism associated with the inhibiting of NOD-like receptor family 3(NLRP3)inflammasome activation.Methods Kunming mice were randomly divided into normal group,model group,Bifendate group,MCC950 group,TFL high dose group and low dose group.Excepting for the normal group,acute liver injury was induced by intraperitoneal injection of 0.2%CCl4-peanut oil solution,then the liver function,morphology and the level of reactive oxygen species(ROS)were detected.The levels of NLRP3 inflammasome-associated protein in liver were determined by western blotting.Results Compared with the normal group,the serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and total bile acid(TBA),and the levels of reactive oxygen species in liver tissues of mice in the model group were significantly increased(P<0.05),and the expression of thioredoxin interacting protein(TXNIP),NLRP3,pro cysteinyl aspartate specific proteinase-1(pro-caspase-1),pro interleukin 1 beta(pro-IL-1β)and interleukin-1β(IL-1β)proteins in liver tissues significantly increased(P<0.05),and the degree of liver tissue damage was severe;Compared with the model group,the treatment of TFL significantly reduced the levels of ALT,AST,TBA in the mice with acute liver injury(P<0.05),decreased levels of liver ROS(P<0.05),attenuated liver morphology damage,and reduced morphological score on HE staining(P<0.05).Meanwhile,high doses of TFL group and MCC950 group significantly lowered the protein levels of TXNIP,NLRP3,pro-caspase-1,pro-IL-1β,and IL-1β(P<0.05).Conclusion TFL attenuates CCl4-induced acute liver injury in mice,the mechanism may lay in the inhibiting of inflammatory responses involved in NLRP3 inflammasome activation.