降脂益肝冲剂对非酒精性脂肪性肝病大鼠肝脏TBP-2mRNA表达的影响

The effect of Jiangzhi Yigan Chongji on TBP-2 mRNA in liver of the rats with nonalclholic fatty liver diseases

目的探讨硫氧还蛋白结合蛋白(TBP-2)与大鼠非酒精性脂肪性肝(NAFLD)的关系,并观察降脂益肝冲剂的疗效,探讨其对肝组织TBP-2mRNA表达的影响。方法72只雄性Wistar大鼠随机分为9组:空白对照组(N组)、实验对照组(M组),实验干预组(D组),每组24只,分别设9、13、17周三个时相点。空白对照组给予普通饲料喂养,实验对照组和实验干预组给予高脂饮食喂养。实验干预组分别于0周、9周、13周末开始给予降脂益肝冲剂,同时空白对照组和实验对照组分别给予等量的生活饮用水灌胃,分别于9周、13周、17周末处死各组大鼠。应用半定量逆转录—聚合酶链反应(RT-PCR)法测定各组大鼠肝细胞硫氧还蛋白结合蛋白(TBP-2)mRNA的表达;HE染色观察肝组织病理变化。结果RT-PCR结果发现在单纯性非酒精性脂肪肝期(M1组)TBP-2的mRNA表达(0.63±0.12)较N1组(1.02±0.10)下降明显(P<0.01),可能为机体的一种保护性应激,以减少TBP-2对Trx的抑制作用,间接增加了机体的抗氧化能力。当疾病继续进展到非酒精性脂肪性肝炎期(M2组和M3组),这种保护性应激能力逐渐减少,TBP-2的mRNA表达增加,但仍然少于同期的正常组即空白对照组(N2,N3)(P<0.01),提示机体仍可通过下调TBP-2的表达以增加了机体的抗氧化能力;N1,N2,N3组相比TBP-2的mRNA表达统计学上无差异;D2组(0.66±0.79)与M2组(0.89±0.10),D3组(0.73±0.12)与M3组(0.90±0.88)相比TBP-2的mRNA表达均有所下降(P<0.01)。药物干预后,肝脏病理改变较实验对照组明显减轻(P<0.01)。结论TBP-2在单纯性非酒精性脂肪性肝和非酒精性脂肪性肝炎期表达下调,可能是机体的一种保护性应激,参与NAFLD的发生、发展。降脂益肝冲剂对NASLD有很好的防治作用,TBP-2是其可能的作用靶点。

Objective To explore the relationship of rat nonalcoholic fatty live（NAFLD） and Thioredoxin-binding protein-2（TBP-2）. To observe the effect of Jiangzhi Yigan Chongji（JYC）on NAFLD rats chronically fed with high-fat diet and explore its effect of expression of TRX -binding protein 2 （TBP-2） mRNA in liver tissue. Methods 72 male Wistar rats were randomly divided into 9 groups（8 rats each group）： blank control group （N）experimental model group（M） and experimental intervention group （D）all of them have three spots 9w、13w、17w.N group fed with with standard diet,M group and D group fed with high-fat diet.The D groups was given JYC by intragastric administration at 0、9、13 weekend,meanwhile,the M and Ngroups were separately given life drinking water by the same means.Then at the end of 9、13、17week,they were put to death. Changes on pathology of liver and hepatocyte TBP-2 expression were observed by hematoxylin and eosin （HE） stain with light microscope and RT-PCR.Results The expression of TBP-2mRNA in liver of every blank control group （N1,N2,N3） kept at similar level without statistical difference（P〉0.05）.The expression of TBP-2mRNA in the liver of M1 group（0.63±0.12） was significantly decreased in comparison with the N1 group（1.02±0.10）（P〈0.01）and increased lightly with the time in the liver of M2 group and M3 group. The expression of TBP-2mRNA in liver of D2 （0.66±0.79）was lower than that in the M2 （0.89±0.10）（P〈0.01）.The expression of TBP-2mRNA in liver of D3 （0.73±0.12）was lower than that in the M3（0.90±0.88）（P〈0.01）. Compared with the M group,the contemporaneous D group shown improvement of the pathology of liver,and with statistical difference （P〈0.01）.Conclusion TBP-2 participate the development of NAFLD and its expression is down-regulation.This is a protective stress for body. JYC have satisfactory therapeutic effects on NAFLD induced with high-fat diet ;and TBP-2 may be the pharmacological therapeutic target.