【摘要】 通过基因芯片技术研究小檗碱、隐丹参酮对胰岛素抵抗卵巢颗粒细胞基因表达的影响。选择猪卵巢颗粒细胞为体外研究对象,利用磷酯酰肌醇-3激酶(PI-3K)特异性抑制剂——沃曼青霉素人工诱导胰岛素抵抗的细胞模型,以0.02g/L小檗碱、隐丹参酮进行干预,继续培养48h,以Trizol法提取总RNA,采用猪全基因表达谱芯片技术筛选出差异表达基因。将筛选得到差异基因输入MAS(Molecule Annotation System)中进行显著性统计分析。结果显示:小檗碱组与沃曼组比较,与细胞凋亡有关的18个基因表达发生显著变化;而隐丹参酮组有12个。这些差异表达的基因涉及TNF家族、bcl-2基因家族、Caspases家族、MAPK信号通路及凋亡传导通路、细胞循环传导通路和tgfb-smad3传导通路。通过对基因芯片上表达水平发生改变基因的分析推测,小檗碱、隐丹参酮诱导卵巢颗粒细胞凋亡是多种途径共同作用的结果,其中Fas和Caspase途径通路上的基因以及细胞周期调控相关的基因可能发挥了主要作用。更多还原

【Abstract】 To study apoptosis related genes of insulin resistance granular cells induced by Berberine and Cryptotanshinone using DNA microarray, ovarian granulose cells from porcine follicles were isolated and cultured in vitro, and insulin resistance was induced by treatment with Wortmannin. The cells were divided into 4 groups, the blank control group cultured with DMSO, the Wortmannin group cultured with wortmannin of 250 nmol/L, the Berberine and Cryptotanshinone group cultured with Wotrmannin of 250 nmol/L as well as the one with Berberine and Cryptotanshinone of 0.02 g/L. After being incubated for 48 h, total RNA was isolated form cells, and the different expressed genes were screened from them by gene chip technique, then analyzed by MAS (Molecule Annotation System) to get differential gene expression profiles and pathways of the four groups. Compared with those in the wortmannin group, there were 18 apoptosis-related genes differently expressed after the treatment of Berberine, while there were 12 with the treatment of Cryptotanshinone. Those genes belong to the Tumor Necrosis Factor (TNF) ligand and receptor super-family, Caspase family, bcl-2 family, cyclin-dependent kinases and inhibitors, MAPK, apoptosis and TGFβ/smads pathways. Results show that the apoptosis process of granulosa cell induced by Berberine and Cryptotanshinone involves many genes and pathways, and the genes among Fas and Casepase pathways and cell cycle regulated genes may play a pivotal role, and thus may promote cell apoptosis and inhibit cell proliferation.更多还原