目的:应用基因表达谱芯片技术筛查3'-大豆苷元磺酸钠干预下脑缺血/再灌注损伤的大鼠大脑组织的差异表达基因,对3'-大豆苷元磺酸钠的脑缺血再灌注损伤保护作用的分子机制的探讨。方法:健康雄性SD大鼠9只,随机分成3组,(A)假手术对照组,(B)脑缺血再灌注损伤模型组,(C)脑缺血再灌注损伤模型+2 mg·kg-13'-大豆苷元磺酸钠组。每组3只,各组于缺血后10 min,A、B组按0.1 m L·100 g-1体重于舌下静脉注射生理盐水,C组从舌下静脉注射2 mg·kg-13'-大豆苷元磺酸钠。缺血90 min,再灌注24 h后,迅速取损伤侧脑组织于液氮中保存。按试剂盒说明,提取样品总RNA。部分总RNA送上海博星基因芯片有限责任公司进行基因芯片检测。结果:全基因表达谱芯片技术分析3'-大豆苷元磺酸钠对脑缺血再灌注损伤的保护作用与以下几个通道有关:NGF、Apoptotic execution phase、Exocytosis of Alpha granule、Amino acid and oligopeptide SLC transporters、Class A/1(Rhodopsin-like receptors)、RNA Polymerase I Transcription Initiation、p75 NTR receptor-mediated signalling、Cell death signalling via NRAGE,NRIF and NADE、Apoptotic cleavage of cellular proteins、Signaling by GPCR、Tachykinin receptors bind tachykinins、Metabolism of amino acids、Response to elevated platelet cytosolic Ca2+、Dissolution of Fibrin Clot、Amino acid transport across the plasma membrane。基因表达谱芯片分析表明3'-大豆苷元磺酸钠能够显著改变1 869个基因mRNA的量,其中mRNA的量上调的有1 084个基因,下调的有785个基因。这些差异表达基因的功能涉及多个方面,如细胞凋亡调控基因、氧化应激基因、炎症相关基因、神经生长基因等。结论:3'-大豆苷元磺酸钠的保护作用是通过多基因、多通道、多层次来实现的。
OBJECTIVE To explore the protective effects of 3′-daidzein sulfonate sodium on chronic hepatic injury induced by carbon tetrachloride(CCl4)in mice and investigate the mechanism about regulating the T lymphocyte subsets.METHODS Healthy Kunming male mice were randomly divided in 5groups:control group,model group,bifendate positive control group(2.5mg·kg-1),low and high dose 3′-daidzein sulfonate sodium groups(0.1and 0.3mg·kg-1).The chronic hepatic injury mice were made by intraperitoneal injection of 10% CCl4 plant oil solution twice a week,and sustained for six weeks.At the same time,the mice were treated with normal saline,bifendate(2.5mg·kg-1)and 3′-daidzein sulfonate sodium(0.1and 0.3mg·kg-1),respectively by ig administration once a day and continued for six weeks.After the last administration,the mice blood and liver were taken.Using automatic biochemical analyzer survey the activity of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in serum.Flow cytometry was used to detect T lymphocyte subsets,enzymes analysis technique was used to observe the liver function(CD3+,CD4+and CD8+),HemateinEosin stain was used to explore the changes in liver morphology.RESULTS The level of ALT and AST in the serum in model group mice increased significantly.While the level of ALT and AST in the serum in 3′-daidzein sulfonate sodium(0.1 or 0.3mg·kg-1,ig)groups mice decreased compared with the model group(P<0.05).And compared with the model group,the ratios of CD3+,CD4-CD8-and CD4+/CD8+ decreased,and the ratios of CD8+ increased in 3′-daidzein sulfonate sodium groups(0.1 and 0.3mg·kg-1,ig;P<0.05).CONCLUSION 3′-daidzein sulfonate sodium have a protective effect on CCl4-induced liver injury mice,and it can result in the changes of T lymphocyte subsets,which may be one of the factors leading to hepatic injury by CCl4.
