紫堇灵对四氯化碳致小鼠急性肝损伤的保护作用及机制的研究

Hepatoprotective Effect of Corynoline on Carbon Tetrachloride-induced Hepatotoxicity in Mice

探讨紫堇灵(Corynoline,COR)对四氯化碳致小鼠急性肝损伤的保护作用及其机制。50只昆明种小鼠随机均分成5组,分别为正常组(Con)、GW9662阻断剂组(GW)、模型组(CCl4)、紫堇灵预处理组(COR)以及紫堇灵与阻断剂GW9662联合处理组(COR+GW),采用腹腔注射0.2%四氯化碳(CCl4)玉米油溶液(10 m L/kg)建造小鼠急性肝损伤模型。20 h后,小鼠脱臼处死后取血清,使用ELISA法检测血液中谷丙转氨酶(ALT)和谷草转氨酶(AST)活性。检测肝脏组织中C反应蛋白(CRP)的含量和肿瘤坏死因子α(TNF-α)的含量。利用HE染色观察肝组织病理学变化,通过Western blot方法检测肝脏组织中过氧化物酶增殖物激活受体γ(PPAR-γ)和NF-κB的蛋白表达。结果显示,紫堇灵能显著降低CCl4性肝损伤所引起的血清中ALT、AST活性升高,明显改善肝组织病理损伤程度;抑制肝脏中炎症因子CRP和TNF-ɑ含量和NF-κB蛋白表达量的升高;有效地拮抗受损肝脏组织中过氧化物酶体增殖物激活受体(peroxisome proliferator-activated receptor,PPAR-γ)蛋白表达量的降低。并且我们发现紫堇灵的这种减轻CCl4性肝损伤的作用几乎全部被GW9662阻断。COR对四氯化碳致小鼠急性肝损伤有一定的保护作用,其机制可能与PPAR-γ和NF-κB信号通路有关。

To investigate the hepatoprotective activities of Corynoline in the mice model of carbon tetrachloride（ CCl4）-induced liver toxicity. In addition,attempts were made to elucidate the possible mechanism of action. Hepatotoxicity was induced in Kunming mice by a intraperitoneal injection（ i. p.） of CCl4,10 mL/kg body weight,diluted with corn oil at a ratio of 1： 500. The corynoline（ COR） was administered once a day for 7 days（ i. p.） as pretreatment at 10 mg/kg·day. The levels of C-reactive protein（ CRP） and tumor necrosis factor-α（ TNF-α） were analyzed to determine the inflammation status. The levels of alanine aminotransferase（ ALT） and aspartate aminotransferase（ AST） were analyzed by ELISA. Liver ultrastructure was observed via optical microscopy. The protein expression degrees of peroxisome proliferator-activated receptor gamma（ PPARγ） and nuclear factor-k-gene binding（ NF-κB） were assayed by Western blot.CCl4-induced hepatotoxicity was manifested by an increase in the levels of ALT,AST,CRP and TNF-ɑ. The histopathological examination of liver sections revealed necrosis and inflammatory reactions. The pretreatment with COR decreased levels of ALT,AST,CRP and TNF-ɑ,decreased the protein expression degrees of NF-κB,and the protein expression degrees of PPARγ,and normalized the hepatic histo-architecture. This study supported the use of COR against hepatotoxicity,and the hepatoprotective effect was mainly through PPARγ and NF-κB signaling passway.