摘要
目的通过基因芯片寻找大鼠激素性股骨头坏死组织中表达变化的基因，尝试探讨激素诱发股骨头坏死的始动因素和分子机制。方法2007年1月至2010年2月，选取20只成年健康Wistar大鼠随机分为对照组和模型组，每组10只。模型组大鼠腹腔注射大肠杆菌内毒素后，于左侧臀肌注射大剂量甲泼尼龙的方法建立激素性股骨头坏死模型；对照组大鼠腹腔注射大肠杆菌内毒素后，于左侧臀肌注射相同剂量生理盐水。6周后取各组大鼠的左侧股骨头标本进行组织病理学观察确认造模成功后，抽提各组大鼠股骨头组织mRNA，经反转录获得大鼠股骨头组织cDNA探针，进行基因芯片检测。检测结果经荧光定量聚合酶链反应（PCR）验证，并用基因本体（geneont^ogy，GO）分类方法对差异表达的基因进行分析。结果与对照组相比，模型组有190条基因的表达存在显著变化，其中52条基因上调表达，138条基因下调表达。这些差异表达基因中102条基因为已知基因（已在GeneBank中登录），88条为未知基因。已知基因根据其生物学功能可被分为：氧化应激、细胞凋亡、信号转导、细胞外基质和脂代谢等基因家族。RT-PCR验证结果与基因芯片检测结果相符。结论激素性股骨头坏死的发生发展是多种因素、多条途径相互作用的复杂的动态过程。实验结果提示，下调表达基因明显多于上调表达基因，说明在此过程中细胞功能发生了简化；差异表达基因中尚有很多未知基因，说明激素性股骨头坏死还有很多未知领域。
Objectives To identify significantly differentially expression genes of steroid-induced femoral head necrosis （SINFH） of rats by gene chip, and to find out the potential factors and molecular mechanisms that oxidative stress originate or strengthen the SINFH. Methods Twenty Wistar rats were divided into experimental group and control group randomly. E. coli endotoxin was given to all rats at a dose of 20 μg/kg body weight by daily i.p. for two times. Then methylprednisolone （40 mg/kg） or saline was daily injected into the left gluteus muscle of the rats in experimental group and control group respectively. Six weeks later, the mRNA was extracted from the femoral head of rats in every group, and the cDNA were obtained by inverse transcript, then carried out microarray detection. The quantitative RT-PCR was used to confirm the result of microarray, and the differentially expressed genes were analyzed for the functional annotation by gene ontology （GO）. Results Compared to the control group, 190 genes in the experimental group were differentially expressed, with 52 up-regulated and 138 down-regulated. Of these genes, 102 are known （have deposited in GeneBank）, while 88 of them are unknown. The known genes can be divided into several families according to their biological functions, such as： oxidative stress, apoptosis, signal transduction, angiogenesis, extracellular matrix, hpid metabolism, and gene transcription related genes. The results of quantitative RT-PCR are consistent with gene-chip results. Conclusions The occurrence of SINFH is a complicated process affected by multiple factors and signaling pathways. Our findings indicate that many genes which are involved in different signaling pathways were differentially expressed between SINFH rats and normal rats.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2011年第10期927-933,共7页
Chinese Journal of Surgery
基金
国家自然科学基金资助项目（30672702）
