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槲皮素拮抗雷公藤甲素诱导的小鼠肝损伤的机制研究

Rotective Effects of Quercetin Against the Triptolide Induced Liver Injury and Relevant Mechanism Study

  • 摘要:
      目的  探究槲皮素(quercetin,QE)对雷公藤甲素(triptolide,TP)诱发肝损伤的拮抗机制。
      方法  取40只C57BL/6小鼠,随机平均分为正常对照组、TP模型组、低剂量QE(20 mg/kg)组和高剂量QE(80 mg/kg)组。低剂量QE组和高剂量QE组进行预处理给药,按0.2 mL/10 g量连续灌胃10 d,2次/d,其余各组灌胃等量生理盐水。末次给药4 h后,除正常对照组外,各组单次灌胃500 μg/kg TP进行造模。造模22 h后取血,处死小鼠,肝切片HE染色观察肝组织病理学改变;检测小鼠血清中谷草转氨酶(AST)和谷丙转氨酶(ALT)活性;测定肝匀浆液中还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)和丙二醛(MDA)水平;采用ELISA方法检测肝匀浆液中白细胞介素(IL)-17、IL-10和IL-6分泌情况;Western blot测定Toll样受体4(TLR4)蛋白表达水平。
      结果  与正常对照组比较,TP模型组肝小叶结构萎缩甚至消失,肝细胞坏死明显,炎性细胞浸润;小鼠血清中ALT、AST和MDA水平异常升高,SOD和GSH水平降低,IL-6和IL-17分泌水平升高,IL-10分泌水平下降,TLR4蛋白水平升高(P < 0.05)。与TP模型组相比,QE组肝组织损伤和炎症细胞浸润减轻,血清中ALT、AST、MDA、IL-6和IL-17水平均下降,TLR4表达水平下调(P < 0.05),且高剂量QE组改变更为明显(P < 0.05,与低剂量QE组比较)。
      结论  槲皮素可通过减轻氧化损伤促进抗氧化,调控细胞因子的分泌,减轻TP诱导的肝损伤。

     

    Abstract:
      Objective   To explore the protective effects of quercetin (QE) on triptolide (TP) induced liver injury and the relevant mechanism.
      Methods   Forty C57BL/6 mice were equally divided into 4 groups, control group, TP model group, 20 mg/kg QE treatment group and 80 mg/kg QE treatment group randomly. The 20 mg/kg and 80 mg/kg QE groups were gastrointestinal administration with QE at the dose of 0.2 mL/10 g for 10 d, twice daily, while other groups were administrated with equivalent normal saline. Four hours post the last dose, animals were gastrointestinal administered with TP at a dose of 500 μg/kg per mouse, except for NS control. All the mice were sacrificed 22 h later, blood and liver tissue samples were collected. The pathologic change of liver tissue was detected by HE staining. The level of aminotransferase (AST) and aspartate alanine aminotransferase (ALT) in serum, and the level of glutathione (GSH), malondialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue homogenates were detected using the commercial kits. The level of interleukin (IL)-17, IL-10 and IL-6 in liver tissue homogenates was measured by ELISA. Hepatic expression of Toll-like receptor 4 (TLR4) was detected by Western blot.
      Results   Compared with the control group, in the TP model group, hepatic lobule structure atrophied and even disappeared, hepatic cell necrosis and inflammatory cell infiltration are obvious. Additionally, in TP model group, serum ALT, AST and MDA levels were significantly increased, SOD and GSH levels were decreased, IL-6 and IL-17 levels were increased, IL-10 levels were decreased, and TLR4 protein levels were increased (P < 0.05). Compared with the TP model group, liver tissue injury and inflammatory cell infiltration were reduced in the QE group, and serum levels of ALT, AST, MDA, IL-6 and IL-17 were all decreased. TLR4 expression was down-regulated (P < 0.05) in both QE groups, and the decease levle was more significant in the high-dose QE group (P < 0.05, compared with the low-dose QE group).
      Conclusion   Quercetin can reduce TP-induced liver injury by reducing oxidative damage, promoting antioxidant and regulating cytokine secretion.

     

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