香叶木素调节NF-κB P65减轻小鼠急性肝损伤

Diosmetin Alleviates Acute Liver Injury in Mice by Regulating NF-κB P65

目的探索香叶木素(Diosmetin, Dios)对D-半乳糖胺/内毒素(D-GalN/LPS)诱导的急性肝损伤小鼠的保护作用和机制.方法 BALB/c小鼠随机分为4组:对照组、加药组、模型组和治疗组.苏木素伊红(HE)染色检测肝组织病理损伤情况;试剂盒检测谷丙转氨酶(ALT)和谷草转氨酶(AST)活力;TUNEL染色检测肝组织细胞凋亡情况;试剂盒检测丙二醛(MDA)、超氧化物歧化酶( SOD)和环氧合酶-2 (COX-2)水平;免疫组化检测肺组织Caspase-3蛋白表达情况;ELISA检测血清炎性因子IL-6、 TNF-α和IL-1β水平;免疫荧光检测P65定位.结果与对照组比较,加药组肝各项肝功能指标无明显变化;模型组细胞出现断裂和红细胞浸润、 ALT和AST水平明显升高,表现出明显肝损伤;Caspase-3、 IL-6、 TNF-α、IL-1β水平、 MDA、 COX-2水平和细胞核P65水平增加, SOD水平减少.而与模型组相比较,治疗组肝损伤明显减轻;治疗组细胞凋亡率比模型组显著降低(P<0. 01);治疗组Caspase-3表达水平随明显减少(P<0. 01);治疗组MDA和COX-2 水平明显减少, SOD水平明显提高(P<0. 01);治疗组炎性因子IL-6、TNF-α、 IL-1β水平显著减少(P<0. 01);细胞核内P65减少.结论 Dios通过抑制NF-κB P65激活LPS诱导的小鼠急性肝损伤.

Objective To investigate the protective effect and mechanism of Diosmetin ( Di-os) on D-galactosamine ( D-GalN )/lipopolysaccharide ( LPS ) induced acute liver injury in mice. Methods BALB/c mice were randomly divided into four groups: control group, Dios group, model group, and treatment group. Hematoxylin eosin ( HE) staining was used to detect the pathological injury of liver tissue, the activities of alanine aminotransferase ( ALT) and aspar-tate aminotransferase ( AST) were detected with the kits, and the apoptosis of liver tissue was de-tected by TUNEL staining. The levels of malondialdehyde ( MDA), superoxide dismutase ( SOD) and cyclooxygenase-2 (COX-2) were detected with the kits, the expression of caspase-3 protein by immunohistochemistry, the serum inflammatory factors IL-6, TNF-α and IL-1β by ELISA, and the localization of p65 by immunofluorescence. Results There were no significant changes in liver func-tion indexes in the Dios group when compared with control group, while in the model group, the cells were broken and erythrocytes infiltrated the tissues, the levels of ALT and AST were signifi-cantly increased, and the liver injury was observed. The levels of caspase-3, IL-6, TNF-α, IL-1 β, MDA, COX-2 and nuclear p65 were significantly increased, while the level of SOD was pro-foundly decreased in model group. Compared with the model group, the liver injury was significantly alleviated, the apoptosis rate and the expression of caspase-3 were significantly reduced in the treat-ment group ( P<0. 01). In the treatment group, the levels of MDA, COX-2, the levels of in-flammatory factors IL-6, TNF-α, IL-1 β, and the level of nuclear p65 were significantly decreased ( P<0. 01), and the level of SOD was obviously increased ( P <0. 01) and. Conclusion Dios alleviates LPS-induced acute liver injury in mice by inhibiting the activation of NF-κB p65.