灵芝多糖对CCl_4诱导的急性肝损伤小鼠的抗炎和保肝活性

目的:研究灵芝多糖对CCl_4诱导的急性肝损伤模型小鼠的抗炎和保肝作用,并探讨其可能机理。方法:小鼠随机均分为6组,即空白对照组,急性肝损伤模型组(体积分数0.1%的CCl_4溶液,20 m L/(kg·d)),灵芝多糖低、中、高剂量组(50、100、150 mg/(kg·d),以体质量计,下同)、联苯双酯组(100 mg/(kg·d))。1周后测定小鼠的体质量、肝质量和肝指数;测定血清中谷丙转氨酶(alanine transferase,ALT)和谷草转氨酶(aspartate transferase,AST)活力及总胆红素(total bilirubin,TBIL)水平;测定肝组织中丙二醛(malondialdehyde,MDA)和还原型谷胱甘肽(glutathione-SH,GSH)水平及一氧化氮合酶(nitric oxide synthase,NOS)活力;测定血清及肝组织炎性因子白细胞介素(interleukin,IL)-1β、IL-18、IL-6及肿瘤坏死因子(tumor necrosis factor,TNF)-α水平;Western blot法测定肝组织中NOD样受体3(NOD-like receptor 3,NLRP3)、凋亡相关微粒蛋白(apoptosisassociated speck-like protein containing CARD,ASC)及胱冬肽酶-1(caspase-1)蛋白表达水平。结果:与空白对照组相比,CCl_4显著提高急性肝损伤模型组小鼠的肝质量和肝指数(P0.05),提高血清ALT和AST活力及TBIL水平,表明造模成功,并且提高IL-1β、IL-18、IL-6、TNF-α水平,提高肝组织MDA、IL-1β水平及NOS活力,提高肝组织NLRP3、ASC及caspase-1的蛋白表达水平,显著降低肝组织GSH水平(P0.05)。与模型组相比,灵芝多糖及联苯双酯显著降低小鼠肝质量和肝指数(P0.05)、降低血清ALT和AST活力及TBIL、IL-1β、IL-18、IL-6、TNF-α水平,降低肝组织MDA、IL-1β水平及NOS活力,下调肝组织NLRP3、ASC及caspase-1的蛋白表达水平,改善肝组织形态学病变,显著提高肝组织GSH水平(P0.05)。结论:灵芝多糖对CCl_4所致急性肝损伤小鼠具有抗炎和保肝作用,其机制可能与抑制自由基脂质过氧化、抑制炎性因子活化及NOS活性有关。

Anti-Inflammatory and Hepatoprotective Effects of Ganoderma lucidum Polysaccharides on Carbon Tetrachloride-Induced Acute Liver Injury in Mice

Objective: To study the anti-inflammatory and hepatoprotective effects of Ganoderma lucidum polysaccharides (GLPS) in mice with acute liver injury induced by carbon tetrachloride (CCl4), and to explore the possible mechanism. Methods: Mice were randomly divided into six groups: normal control group, acute liver injury model group （0.1% (V/V) CCl4, 20 mL/(kg bw·d), low, medium and high-dose GLPS groups (50, 100 and 150 mg/(kg bw·d)), and bifendate group (100 mg/(kg bw·d)). After administration for 7 continuous days, the body weight, liver weight, liver index, alanine transferase (ALT) and aspartate transferase (AST) activities, and total bilirubin (TBIL) level in serum were detected, and the malondialdehyde (MDA) and glutathione-SH (GSH) levels, and nitric oxide synthase (NOS) activity in liver tissue were also analyzed. The levels of interleukin (IL)-1β, IL-18, IL-6 and TNF-α in serum and IL-1β in liver tissue were analyzed by commercial enzyme linked immunosorbent assay (ELISA) kits. The protein expression levels of NLRP3, ASC and caspase-1 in liver tissue were evaluated by Western blot. Results: Compared with the normal control group, CCl4 significantly increased liver weight and liver index, improved serum ALT and AST activities, enhanced the levels of TBIL, IL-1β, IL-18, IL-6 and TNF-α in serum and MDA, IL-1β and NOS in liver tissue, up-regulated the protein expression of NLRP3, ASC and caspase-1 in liver tissue, and remarkably reduced GSH level in liver tissue in the model group. Compared with the acute liver injury model group, GLPS and bifendate notably inhibited ALT and AST activities, decreased liver weight and index, as well as TBIL, IL-1β, IL-18, IL-6 and TNF-α in serum and MDA, IL-1β and NOS in liver tissue, remarkably enhanced GSH levels, down-regulated the protein expression NLRP3, ASC and caspase-1 in liver tissue, notably ameliorated the morphological changes of liver tissue. Conclusion: GLPS has a significant anti-inflammatory and hepatoprotective effect on CCl4-induced liver injury in mice. The possible mechanism may be related to the inhibition of free radical lipid peroxidation, NOS activity and the activation of liver inflammatory factors.