大麻二酚对小鼠肝纤维化抗氧化作用

马润; 舒远辉; 谢娜; 周艳; 项一宁; 王豫萍

doi:10.11847/zgggws1135692

摘要:

目的探讨大麻二酚（CBD）对四氯化碳（CCL₄）构建小鼠肝纤维化模型的抗氧化作用。

方法 C57BL/6J雄性小鼠随机分为5组，每组8只，依次为对照组，模型组，低、高剂量CBD组（4、8 mg/kg），秋水仙素组（0.2 mg/kg）。对照组小鼠腹腔注射花生油，剩余小鼠按照5 mL/kg注射30 % CCL₄，每周2次，连续10周。CBD组和秋水仙素组在造模的同时每次注射相应剂量药物，对照组和模型组注射生理盐水。自动生化分析仪检测血清丙氨酸氨基转移酶（ALT）；天狼星红染色检查肝组织胶原沉积情况；试剂盒检测肝组织氧化应激相关成分超氧化物歧化酶（SOD）、还原型谷胱甘肽（GSH）、丙二醛（MDA）；蛋白印迹（WB）检测肝组织还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶亚基gp91phox、核转录相关因子（Nrf2）表达情况。

结果与模型组比较，低、高剂量CBD组和秋水仙素组肝指数分别为（4.71 ± 0.34）%、（4.68 ± 0.20）%、（4.24 ± 0.40）%、血清ALT含量分别为（79.38 ± 14.79）、（87.13 ± 9.40）、（77.88 ± 3.76）U/L明显降低（P < 0.05），胶原沉积显著减少（P < 0.05），肝组织SOD活力分别为（259.90 ± 36.05）、（223.13 ± 37.49）、（246.39 ± 53.49）U/mgprot、GSH含量分别为（63.07 ± 16.71）、（100.38 ± 22.44）、（96.39 ± 19.40）nmol/mgprot明显增高（P < 0.05），而MDA含量分别为（2.48 ± 0.43）、（2.74 ± 0.88）、（2.84 ± 0.87）nmol/mgprot显著降低（P < 0.05），gp91蛋白表达降低（P < 0.05），Nrf2蛋白表达增加（P < 0.05）。

结论 CBD对CCL₄诱导的小鼠肝纤维化有一定的保护作用，其作用机制可能与调节gp91、Nrf2蛋白表达，减少自由基、抑制脂质过氧化相关。

Abstract:

Objective To explore antioxidant effect of cannabidiol (CBD) in mice with carbon tetrachloride (CCl4)-induced liver fibrosis.

Methods Forty male C57BL/6J mice were divided into five groups: a control and a model group, two CBD groups (4 mg/kg and 8 mg/kg) and a colchicine (0.2 mg/kg) group. Intraperitoneal administration of carbon tetrachloride (CCl4) at the dosage of 5 mL/kg body weight was performed for all the mice twice a week for consecutive ten weeks except for those of the control group with intraperitoneal injection of peanut oil; meanwhile, the mice of CBD and colchicine groups were intraperitoneally injected with corresponding dose of medication each time and the control mice with saline. Serum alanine aminotransferase (ALT) was measured with an automatic biochemical analytic instrument; collagen deposition of liver tissue was examined with sirius red staining; superoxide dismutase (SOD), reduced glutathione (GSH) and malondialdehyde (MDA) were measured using commercial kits; gp91/Nrf2 in liver specimens was assessed with Western blot.

Results In the mice of treated with low/high CBD and colchicine, significantly lower liver index (4.71 ± 0.34%/4.68 ± 0.20% and 4.24 ± 0.40%), serum ALT (79.38 ± 14.79/87.13 ± 9.40 and 77.88 ± 3.76 U/L), MDA in liver tissues (2.48 ± 0.43/2.74 ± 0.88 and 2.84 ± 0.87 nmol/mgprot) and significantly increased SOD (259.90 ± 36.05/223.13 ± 37.49 and 246.39 ± 53.49 U/mgprot) and GSH (63.07 ± 16.71/100.38 ± 22.44 and 96.39 ± 19.40 nmol/mgprot) in liver tissues were detected compared with those in the model rats (P < 0.05 for all); obviously ameliorated collagen deposition, down-regulated protein expression of gp91 but up-regulated and protein expression of nuclear factor-erythroid 2-related factor-2 (Ntf2) were also observed in liver tissues of the low/high CBD and colchicine groups (all P < 0.05).

Conclusion The results suggeste that CBD has protective effect on CCl4-induced liver fibrosis in mice, probably via adjusting protein expression of gp91 and Nrf2, clearing free radicals and inhibiting the occurrence of lipid peroxidation.

大麻二酚对小鼠肝纤维化抗氧化作用

Antioxidant effect of cannabidiol in mice with liver fibrosis