杠板归总黄酮对抗结核药物致肝损伤小鼠的保护作用

Protective effect of total flavonoids extracted from Polygonum perfoliatum L.on the liver injury caused by anti-tuberculosis drugs in mice

目的研究杠板归总黄酮(TFP)对抗结核药物致肝损伤小鼠的保护作用及核转录因子-E2相关因子2-抗氧化反应元件(Nrf2-ARE)信号通路的影响及其相关机制。方法将72只小鼠随机分为空白组、模型组、对照组和低、中、高剂量实验组。空白组灌胃等体积生理盐水,每天1次,共给药30 d,模型组灌胃异烟肼100 mg·kg^(-1)和利福平100 mg·kg^(-1)进行造模,其他同空白组一致,对照组灌胃100 mg·kg^(-1)水飞蓟素;低、中、高剂量实验组分别灌胃150,300和600 mg·kg^(-1)TFP。末次给药后,采集肝组织,以苏木素-伊红(HE)染色观察肝组织病理学变化;眼球取血,检测血清中肝功能活性指标、氧化应激指标;以蛋白质印迹法检测肝组织Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)和Nrf2-ARE信号通路相关蛋白表达量。结果模型组和低、中、高剂量实验组的血清谷丙转氨酶(GPT)分别为(282.68±26.87),(243.60±24.16),(164.72±9.64),(64.99±10.22)U·L^(-1),谷草转氨酶(GOT)分别为(302.99±23.64),(263.46±26.30),(181.07±15.33),(86.28±13.73)U·L^(-1),血清磷酸酶(AKP)分别为(22.81±1.24),(17.95±0.99),(14.20±0.97),(9.93±1.29)U·L^(-1),丙二醛(MDA)分别为(1.52±0.14),(1.21±0.15),(1.05±0.18),(0.90±0.08)nmol·mg^(-1),超氧化物歧化酶(SOD)分别为(69.45±5.94),(82.64±0.77),(101.58±10.60),(121.01±15.45)U·mg^(-1),GSH-Px分别为(1.85±0.20),(2.16±0.29),(3.04±0.37),(3.53±0.24)μmol·L^(-1),实验组与模型组比较,差异均有统计学意义(均P<0.05)。与模型组相比,实验组肝组织中Bcl-2、Nrf2、HO-1和Bax蛋白表达,差异均有统计学意义(均P<0.05)。结论杠板归总黄酮对抗结核药物导致的小鼠肝损伤起到保护作用,其机制可能与Nrf2-ARE信号通路有关。

Objective To explore the protective effect of total flavonoids(TFP)of anti-tuberculosis drugs in mice with liver injury and the influence of nuclear transcription factor-E2 related factor 2-antioxidant response element(Nrf2-ARE)signaling pathway and related mechanisms.Methods The 72 mice were randomly divided into blank group,model group,control group and low,medium and high dose experimental groups.Blank group was given equal volume of normal saline once a day for 30 d.Model group was given isoniazid 100 mg·kg^(-1) and rifampicin 100 mg·kg^(-1) for modeling,others were the same as blank group,control group was given 100 mg·kg^(-1) silymarin by gavage;low,medium and high dose experimental groups were given 150,300 and 600 mg·kg^(-1) TFP by gavage respectively.After the last administration,the liver tissues were collected,and hematoxylin-eosin(HE)staining was used to observe the pathological changes liver tissues;blood was taken from eyeballs to detect liver function activity indicators and oxidative stress indicators in serum;Western blot experiments was used to detect liver tissues Bcl-2 related X protein(Bax),B cell lymphoma/leukemia-2(Bcl-2)and Nrf2-ARE signaling pathway related protein.Results The glutamic-pyruvic transaminase(GPT)in model group and low,medium and high dose experimental groups were(282.68±26.87),(243.60±24.16),(164.72±9.64),(64.99±10.22)U·L^(-1),glutamic-oxaloacetic transaminase(GOT)were(302.99±23.64),(263.46±26.30),(181.07±15.33),(86.28±13.73)U·L^(-1),alkaline phosphatase(AKP)were(22.81±1.24),(17.95±0.99),(14.20±0.97),(9.93±1.29)U·L^(-1),malondialdehyde(MDA)were(1.52±0.14),(1.21±0.15),(1.05±0.18),(0.90±0.08)nmol·mg^(-1),superoxide dismutase(SOD)were(69.45±5.94),(82.64±0.77),(101.58±10.60),(121.01±15.45)U·mg^(-1),glutathione peroxidase(GSH-Px)were(1.85±0.20),(2.16±0.29),(3.04±0.37),(3.53±0.24)μmol·L^(-1).Compared with model group,the above indexes in experimental groups were all with significant difference(all P<0.05).Compared with model group,the expressions of Bcl-2,Nrf2,HO-1 and Bax protein in the liver tissue of experimental group were all with statistically difference(all P<0.05).Conclusion The TFP have a protective effect on the liver injury of mice caused by anti-tuberculosis drugs,and the mechanism may be related to Nrf2-ARE signaling pathway effect.