樟芝多糖保护小鼠急性肝损伤的机制研究

Protective Mechanism of Polysaccharides from Anoderma Camphora on Acute Liver Injury in Mice

目的研究樟芝多糖保护小鼠急性肝损伤的机制。方法通过D-氨基半乳糖构建急性肝损伤小鼠,C57BL/6小鼠分为对照组、模型组、樟芝多糖高剂量组、樟芝多糖低剂量组,樟芝多糖高、低剂量组每日分别给予50,25mg·kg-1的樟芝多糖2次,对照组和模型组给予等体积的生理盐水。给药7d后紫外比色法测试盒检测大鼠肝组织中谷丙转氨酶(ALT)、谷草转氨酶(AST)水平;TBA法试剂盒检测肝组织中丙二醛(MDA)的水平;ELISA试剂盒检测肝组织中白介素-6(IL-6)的表达;Western-Blot检测肝脏组织中NLRP-3、白介素-1β(IL-1β)、pro-caspase-1和caspase-1的表达;HE染色观察小鼠肝脏病理;RT-qPCR检测肝脏组织中Bcl-2、Bax、caspase-3、caspase-1的mRNA表达。结果与模型组相比,樟芝多糖可以明显改善急性肝损小鼠的肝脏病理,降低ALT、AST的表达以及MDA和IL-6的表达(P<0.01),肝脏组织中NLRP-3、IL-1β以及pro-caspase-1和caspase-1的表达相比模型组显著降低(P<0.05),Bax、caspase-3、caspase-1的mRNA表达相比模型组显著降低(P<0.01),Bcl-2显著升高(P<0.01)。结论樟芝多糖对于小鼠急性肝损伤有着很好的保护作用,其作用机制与炎性小体NLRP-3及其相关炎症因子的抑制有关。

OBJECTIVE To study the mechanism of protect effect of polysaccharides from Anoderma camphora on the acute liver injury in mice.METHODS Acute liver injury mice were constructed by D-galactosamine.C57BL/6 mice were divided into control group,model group,high and low dose group of Anoderma camphora polysaccharides.High and low dose group of Anoderma camphora polysaccharides were given 50,25 mg·kg-1 Anoderma camphora polysaccharides for 2 times daily,the control group and model group were given saline measurement.After administration of 7 d,UV colorimetric test box was used to detect alanine aminotransferase(ALT),aspartate aminotransferase(AST);TBA test box was used to detect malondialdehyde(MDA)and ELISA method detected interleukin-6(IL-6)expression in liver tissues of rats;Western-blot was used to detect the expression of NLRP-3,interleukin-1β(IL-1β),pro-caspase-1 and caspase-1 in liver tissues;HE staining was used to observe the liver pathology of mice.The expression of Bcl-2,Bax,caspase-3 and caspase-1 mRNA in liver tissues was detected by RT-qPCR.RESULTS The polysaccharide from Anoderma camphora could obviously improve the liver pathology in the liver lesion mice,the expression of ALT and AST and the expression of MDA and IL-6 were significantly lower than those of model group(P<0.01).The expression of NLRP-3,IL-1β,pro-caspase-1 and caspase-1 in liver tissue was significantly lower than that in model group(P<0.05).The expression of Bax,caspase-3 and caspase-1 mRNA was significantly lower than that in model group(P<0.01),and Bcl-2 increased significantly(P<0.01).CONCLUSION Polysaccharide from Anoderma camphora has a good protective effect on acute liver injury mice.The mechanism is related to the inhibition of inflammatory body NLRP-3 and related inflammatory factors.