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

关键词:  樟芝多糖  炎性小体NLRP-3  急性肝损伤

Key words:  polysaccharide from Anoderma camphora  inflammatory corpuscle NLRP-3  acute liver injury

摘要:

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50，25 mg·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7 d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。

Abstract:

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.