丹参对奶牛脂肪变性肝细胞NF-κB和CYP450表达的影响  

Effects of salvia miltiorrhiza on NF-κB and CYP450 expressions in steatosis hepatocytes of dairy cows

作　　者：王安忠[1] 路丽芹 李鹏飞[1] 秦建华[1] 马玉忠[1] 

机构地区：[1]河北农业大学动物医学院,河北保定071001

出　　处：《饲料工业》2014年第21期50-53,共4页Feed Industry

基　　金：河北省自然科学基金"细胞色素酶P450与奶牛脂肪肝形成关系的研究[C2011204076]";河北省现代农业产业技术体系奶牛产业创新团队资助

摘　　要：研究丹参对脂肪变性奶牛肝细胞的作用机制。采用胶原酶两步灌流法分离培养奶牛肝细胞,不同浓度DL-乙硫氨酸处理,油红染色观察其脂滴数量,MTT法测定肝细胞的活性,生化法检测TG含量。用不同浓度的丹参处理变性肝细胞后,生化法检测ALT、AST、TG、SOD、MDA的含量或活性,ELISA法测定TNF-α的含量,Western blot法检测NF-κB和CYP450的蛋白表达水平。筛选出2 mmol/l的DL-乙硫氨酸作用24 h作为体外造模的最佳条件。用丹参处理后,能降低ALT、AST、TG和MDA的活性或含量,抑制TNF-α的释放,增加SOD活性。随丹参浓度增加和处理时间的延长,CYP450的表达呈逐渐降低的趋势。24 h时,丹参对NF-κB的表达无明显影响;48 h时,随丹参浓度的增加,NF-κB的表达呈逐渐降低的趋势。丹参对奶牛脂肪肝有一定的治疗作用,其作用机制与促进脂质代谢、抑制肿瘤细胞坏死因子、抗脂质过氧化及降低NF-κB和CYP450的表达密切相关。To investigate salvia miltiorrhiza acting meehanism on steatosis hepatocytes in dairy, cows. The hepatocytes in dairy cows were isolated by two-step collagenase perfnsion method, and treated with Dl,-ethionine. The lipid droplets were observed by oil red staining. Tile hepatocytes viability was measured by MTT method. TG in lysis hepatocytes was determined. The steatosis hepatocytes were Irealed with gradienl concenh＇ation of salvia miltiorrhiza. The contents of ALT, AST, TG, SOD, MDA, TNF-α in the hepalocyles were detected. The expressions of NF-KB and CYP450 were determined by Western biol. 2 mmol/l DL-ethionine treated the hepatocyles for 24 h was the optimal condition for steatosis model in vitro. With the treatment of salvia miltiorrhiza, the contents of AI,＇F, AST, MDA and TG decreased. the release of TNF-α was inhibited, and the activity of SOD increased. The expression of CYP450 in hepatocytes decreased gradually. The NF-KB had no significant expression for 24 h treatment, but the expression of NF-KB decreased gradually with the concentration increasing of salvia mihiorrhiza for 48 h treatment. These findings suggest that salvia miltiorrhiza has some therapeutic effect on dairy cow fatty liver, it maybe relate to promote lipid metabolism, inhibit tumor cell necrosis factor, oppose lipid peroxidation and decrease the expressions of NF-KB and CYP450.

关 键 词：丹参 肝细胞 脂肪变性 NF-ΚB CYP450 

分 类 号：S816.32[农业科学—饲料科学]