基于PCR array技术探讨高脂血症大鼠肝脏AS信号通路相关基因mRNA表达

The mRNA expression of the AS signaling pathway related genes in the liver of hyperlipidemia rats by PCR array technology

目的利用PCR array技术检测高脂血症大鼠肝脏动脉粥样硬化(AS)信号通路相关基因mRNA水平,初步揭示上述基因表达变化与高脂血症发生的关系。方法 SPF级健康雄性SD大鼠30只,随机分为空白对照组和高脂血症组,高脂血症组给予高脂饲料喂饲造模。全自动生化分析仪检测血清胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDLC)、高密度脂蛋白胆固醇(HDL-C),HE染色观察肝脏形态变化,油红O染色观察肝脏脂质沉积,提取各组大鼠肝脏总RNA,应用AS信号通路PCR芯片检测高脂血症大鼠肝脏AS信号通路相关基因mRNA表达。结果与空白对照组相比,高脂血症组大鼠血清HDL-C水平显著降低,TC和LDL-C水平显著升高,肝细胞形成大量脂质沉积。PCR array技术分析发现:与空白对照组相比,高脂血症组大鼠上调≥2倍的基因49个,占55%,如Serpinb2(PAI-2)、Il2、Fga、Il5、Il4、Tnf、Cd44、Bid、Vwf、Mmp3、Icam1、Apob等基因;下调≥2倍的基因20个,占22%,如Tgfb1、Vegfa、Lpl、Sod1、Fas、Pparg等基因,涉及应激反应、细胞凋亡、凝血、粘附分子、脂质转运与代谢、细胞生长和增殖以及转录调节等方面。结论肝脏AS信号通路相关基因mRNA表达变化可能与高脂血症发生有关。

Objective To test the mRNA expression of AS signaling pathway related genes in the liver of hyper- lipidemia rat by the method of PCR array, and to explore the relationship between the above - mentioned changes in gene expression and hyperlipidemia. Methods SD rats （30 males） were randomly divided into two groups： control group and model group. The rats in model group were fed with high fat diet. The content of TG, TC, HDL - C and LDL - C in serum were detected by automatic biochemical analyzer. HE and oil red staining meth- od were used to observe the liver injury. Total RNA was extracted from liver in each group. PCR array was used to test the AS signaling pathway related genes in the liver. Results In model group the level of TC and LDL - C increased while the level of HDL - C decreased significantly compared to control group. Oil red staining indica- ted that obvious lipid deposition were formed in the liver cells in model group. DNA microarray showed that in model group the expression level of 49 （55%） genes （e. g. Serpinb2 （PAI- 2）, I12, Fga, I15, I14, Tnf, Cd44, Bid, Vwf, Mmp3, Icaml, Apob, etc. ） increased by more than 2 -fold compared to control group; and the expression level of 20 （22%） genes （ e. g. Tgfbl, Vegfa, Lpl, Sodl, Fas, Pparg, etc. ） decreased by more than 50% compared to control group. Conclusion The mRNA expression of AS related genes may relate to the hyperlipidemia.