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本研究用Sprague-Dawley大鼠建立肝脏缺血/再灌注损伤模型,探讨木犀草素预处理对大鼠肝脏缺血/再灌注损伤的保护作用及其机制,并观察血红素氧合酶-1(heme oxygenase-1,HO-1)活性变化对肝缺血/再灌注损伤的影响。将大鼠随机分为正常组、模型组、木犀草素组、木犀草素+锌原卟啉(HO-1抑制剂)组及血红素组,每组8只。正常组、模型组和血红素组大鼠每日喂以正常饲料,木犀草素组和木犀草素+锌原卟啉组大鼠每日喂以补充木犀草素(200mg/kg)的饲料,喂养4周。木犀草素+锌原卟啉组及血红素组大鼠于实验前6h分别皮下注射锌原卟啉(25μmol/kg)和血红素(20μmol/kg)。除正常组大鼠肝脏不作缺血处理外,其余各组大鼠肝门夹闭阻断血流缺血60min之后,再灌注60min。测定血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)及超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)含量;检测肝组织SOD活性、MDA含量及HO-1活性及蛋白表达情况;HE染色观察肝脏形态结构变化。结果显示:与模型组相比,木犀草素组和血红素组大鼠血清ALT、AST活性及MDA含量均明显下降,SOD活性显著增强(P<0.01);肝组织中MDA含量均明显下降,SOD及HO-1活性显著增强,HO-1蛋白表达量增加(P<0.01);木犀草素+锌原卟啉组大鼠血清ALT、AST活性含量较木犀草素组明显升高(P<0.01),肝组织SOD及HO-1活性下降,MDA含量升高(P<0.01)。组织切片显示模型组肝细胞肿胀,小叶结构紊乱;木犀草素和血红素处理组肝细胞轻度肿胀,小叶结构清晰。结果提示,木犀草素通过降低MDA含量、提高SOD活性及HO-1蛋白表达,改善肝脏的抗氧化能力,减轻缺血/再灌注损伤。
In this study, a model of liver ischemia / reperfusion injury was established in Sprague-Dawley rats to explore the protective effect and mechanism of luteolin preconditioning on hepatic ischemia / reperfusion injury in rats. Heme oxygenase-1 (heme oxygenase-1, HO-1) on hepatic ischemia / reperfusion injury in rats. The rats were randomly divided into normal group, model group, luteolin group, luteolin + zinc protoporphyrin (HO-1 inhibitor) group and heme group, 8 rats in each group. Rats in normal group, model group and heme group were fed daily with normal diet, luteolin group and luteolin + zinc protoporphyrin group rats were fed daily with luteolin (200mg / kg) Feeding for 4 weeks. Luteolin + zinc protoporphyrin group and heme group rats were injected subcutaneously with zinc protoporphyrin (25μmol / kg) and heme (20μmol / kg) 6 h before the experiment. Except the normal group, the liver of rats was not subjected to ischemic treatment, and the rest of the hepatic vessels in each group were occluded for 60 minutes after ischemia and reperfused for 60 minutes. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD) and malondialdehyde (MDA) Content of liver tissue were measured; SOD activity, MDA content, HO-1 activity and protein expression in liver tissue were detected; morphological changes of liver were observed by HE staining. The results showed that compared with the model group, the activities of ALT, AST and MDA in the luteolin group and the hemoglobin group decreased significantly and the SOD activity increased significantly (P <0.01); the content of MDA in the liver tissue decreased significantly, SOD and HO-1 activity were significantly increased and HO-1 protein expression was increased (P <0.01). The contents of ALT and AST in serum of luteolin + zinc protoporphyrin group were significantly higher than those of luteolin group (P < 0.01). The activities of SOD and HO-1 in liver tissue decreased and MDA content increased (P <0.01). Tissue sections showed hepatocyte swelling and lobular structure disorder in the model group. The hepatocytes in luteolin and heme treatment groups had slight swelling and the structure of leaflets was clear. The results suggest that luteolin can reduce the content of MDA, increase the activity of SOD and the expression of HO-1, improve the anti-oxidation ability of the liver and reduce the ischemia / reperfusion injury.