开心果总黄酮预处理对急性肝损伤小鼠肝组织的保护作用及机制

Protective effect of total flavonoids of Pistacia vera L. on liver tissues of mice with acute liver injury

目的观察开心果总黄酮预处理对急性肝损伤小鼠肝组织的保护作用,并探讨其作用机制。方法取50只健康昆明种小鼠,随机分为正常组、模型组及开心果总黄酮低、中、高剂量组各10只。开心果总黄酮低、中、高剂量组分别给予100、200、400 mg/kg开心果总黄酮灌胃,正常组和模型组给予等量蒸馏水灌胃,均1次/d,连续7d。末次给药1 h后,正常组腹腔注射花生油溶液,模型组和开心果总黄酮各剂量组腹腔注射CCl_4花生油溶液建立急性肝损伤模型。造模后8 h,小鼠摘眼球取血,使用全自动生化分析仪测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,酶标仪检测血清白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)水平;处死小鼠,摘取肝脏和脾脏并称重,计算肝脏指数和脾脏指数;取肝组织,HE染色法观察肝脏组织病理学变化;使用全自动生化分析仪测定肝组织中超氧化物歧化酶(SOD)、一氧化氮(NO)、丙二醛(MDA)、谷胱甘肽(GSH)水平。结果与正常组比较,模型组血清ALT、AST及IL-6、TNF-α、IL-1β、IFN-γ水平均升高;与模型组比较,开心果总黄酮中、高剂量组上述指标均降低(P <0. 05或<0. 01)。与正常组比较,模型组肝脏指数、脾脏指数均升高;与模型组比较,开心果总黄酮中、高剂量组肝脏指数、脾脏指数均降低(P均<0. 05)。正常组肝小叶结构完整,肝细胞无明显变性坏死;模型组肝小叶结构和肝细胞排列紊乱,肝细胞水肿、坏死,存在炎性细胞浸润;与模型组比较,开心果总黄酮组中、高剂量组肝小叶结构基本完整,肝细胞水肿减轻,肝细胞坏死减少,炎性细胞少见。与正常组比较,模型组肝组织中MDA、NO水平升高,SOD、GSH水平降低(P均<0. 05);与模型组比较,开心果总黄酮中、高剂量组肝组织中MDA、NO水平降低,SOD、GSH水平升高(P均<0. 05)。结论开心果总黄酮对CCl_4诱导的小鼠急性化学性肝损伤具有较好的防治作用,其机制可能与抑制自由基脂质过氧化、抑制炎性因子活化有关。

Objective To study the protective effect of total flavonoids of Pistacia vera L.on the liver tissues of mice with carbon tetrachloride(CCl 4)-induced acute liver injury and its mechanism.Methods Fifty healthy adult mice were randomized into five groups:the control group,model group,different doses groups of total flavonoids of Pistacia vera L.(100,200,and 400 mg/kg,low-dose,medium-dose and high-dose groups),each mouse was intragastrically administrated for 7 days.One hour after the last administration,except for the control group,the acute liver injury models induced by CCl 4 were established by intraperitoneal injection of 0.1%mL/CCl 4 peanut oil solution(0.1 mL/10 g body weight)in the other mice.At 8 h after the model establishment,the blood samples were taken from the eyeballs of mice,the serum levels of alanine aminotransferase(ALT)and oxaloacetic aminotransferase(AST)were measured by automatic biochemical analyzer,and the serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α)were detected by using microplate reader.After that,the mice were killed and the liver and spleen were taken out and weighed to measure the index of liver and spleen,the liver tissues were collected and observed by HE staining;the levels of superoxide dismutase(SOD),nitric oxide(NO),malondialdehyde(MDA),and glutathione(GSH)in the liver tissues were measured by automatic biochemical analyzer.Results Compared with the normal group,the levels of serum ALT,AST,IL-6,TNF-α,IL-1β,and IFN-γincreased in the model group;compared with the model group,all the above indexes decreased in the medium-dose and high-dose groups of total flavonoids of Pistacia vera L.(P<0.05 or<0.01).Compared with the normal group,the liver index and spleen index of the model group increased,and the liver index and spleen index of the medium-dose and high-dose groups of total flavonoids of Pistacia vera L.decreased(all P<0.05).The structure of hepatic lobules in the normal group was intact,and hepatocytes did not have degeneration and necrosis;the structure of hepatic lobules and hepatocytes were arranged disorderly;hepatocytes edema,necrosis and inflammatory cell infiltration were found in the model group.Compared with the model group,the structure of hepatic lobules was basically intact,liver cell edema was lightened,liver cell necrosis was reduced,and inflammatory cells were rare in the medium-dose and high-dose groups of total flavonoids of Pistacia vera L..Compared with the normal group,the levels of MDA and NO in the liver tissues of model group increased,while the levels of SOD and GSH decreased.Compared with the model group,the levels of MDA and NO decreased in the medium-dose and high-dose groups of total flavonoids of Pistacia vera L.,and the levels of SOD and GSH increased(all P<0.05).Conclusion Total flavonoids of Pistacia vera L.has a good preventive and therapeutic effect on the liver tissues of mice with CCl 4-induced acute liver injury by inhibiting the free radical lipid peroxidation and inhibiting the activation of inflammatory factors.