鞣花酸对AKT基因转染诱导小鼠脂肪性肝病中炎症及氧化应激的影响

Effects of ellagic acid on inflammation and oxidative stress induced by AKT gene transfection in mice with fatty liver disease

研究鞣花酸(ellagic acid,EA)对AKT基因转染诱导小鼠脂肪性肝病中炎症及氧化应激的影响。将20只雌性FVB小鼠随机分为正常对照组,模型组,EA低、高剂量给药组(150,300 mg·kg^(-1)·d^(-1))。模型组及EA给药组采用高压尾静脉转染法转入AKT质粒,EA连续给药5周,模型组及正常对照组给予等量生理盐水。取各组小鼠肝组织,采用HE染色法,油红O染色法观察小鼠肝组织病理学改变;检测各组小鼠肝功能以及血清和肝组织中的MDA和SOD水平;实时荧光定量PCR(RT-q PCR)法检测NF-κB,TNF-αmRNA的表达水平;Western blot法、免疫组织化学法检测肝组织中NF-κB,TNF-α,IL-6,COX-2等蛋白的表达水平。结果显示,与正常对照组相比,模型组小鼠血清中AST,ALT水平升高,血清及肝组织中MDA水平升高,SOD水平降低,组织病理学结果显示肝组织脂肪化明显,损伤严重。EA给药后,肝脂肪化显著减轻,AST,ALT,MDA水平降低,SOD水平升高,肝组织中NF-κB,TNF-α,IL-6,COX-2等表达水平降低。以上结果表明,EA对AKT基因转染诱导的小鼠脂肪性肝病具有治疗作用,其作用机制可能与抑制NF-κB,TNF-α,IL-6,COX-2等炎症相关因子的表达、抗氧化应激损伤有关。

To study the effects of ellagic acid(EA)on inflammation and oxidative stress in mice with fatty liver disease induced by AKT gene transfection,the 20 female FVB mice were randomly divided into normal control group,model group and ellagic acid administration group(150,300 mg·kg^-1·d^-1)(n=5).EA experimental groups and model group were using a high pressure into the tail vein transfection plasmid AKT.The next day,EA was started to administered continuously for 5 weeks after the AKT gene transfection,while the model group and the normal control group were given the same amount of saline.After the administration,the liver tissue and serum of mice were taken.HE and oil red O staining were using to observe the histopathological changes in liver;liver function to detect the serum and liver tissue as well as MDA and SOD levels;real-time quantitative PCR(RT-qPCR)was used to measure the mR-NA expression of NF-κB and TNF-α;Western blot and immunohistochemistry were used to measure the expression of NF-κB,TNF-αand COX-2 in liver tissue.Results show that after AKT gene transfection,the model group had significant increase in the serum levels of AST,ALT,elevated the levels of MDA and decreased the levels of SOD in serum and liver tissue,aggravated histopathology degeneration and Liver inflammation,and significantly higher expression of NF-κB,TNF-α,IL-6,COX-2 and other inflammatory-related factors in liver tissue.EA administration group significant reductions in the serum levels of AST,ALT,and improved in hepatocyte fatty degeneration and liver inflammation,lower the levels of MDA and increased the levels of SOD in serum and liver tissue,and significant reductions in the expression of NF-κB,TNF-α,IL-6 and COX-2 in liver tissue.These results suggest that EA has obvious anti-inflammatory effect and inhibits oxidative stress and EA has a significant therapeutic effecton AKT gene inducing fatty liver,and the mechanism possibly by inhibiting inflammatory factors of NF-κB,TNF-α,IL-6,COX-2 and anti-oxidative stress-related.