摘要目的 使用基因芯片技术分析糖尿病早期大鼠视网膜血管凋亡相关基因表达概况.方法 腹腔注射链脲佐菌素(STZ)制备糖尿病大鼠模型.在血糖升高后的第6周分别处死正常组大鼠和糖尿病组大鼠各10只,提取20只眼的视网膜血管,一步法提取总RNA.使用(α-32P)脱氧腺苷酸(dATP)标记样品制作探针,与含有1176个基因的尼龙膜芯片进行杂交.使用计算机软件对所获结果进行相关分析.选择3个差异表达的基因进行逆转录聚合酶链反应(RT-PCR)验证.结果 糖尿病大鼠第6周,136个基因具有差异表达.占检测基因总数的11.5%.其中.表现为上调的基因90个,占7.6%;表现为下调的基因46个,占3.9%.差异表达涉及多种功能的多个基因.与凋亡信号传导通路相关的72个基因中.有15个出现了表达的差异.表达上调的基因包括肿瘤坏死因子(TNF)家族中Fas相关的死亡域(FADD)、TNF受体家族成员12(TNFRSF12)、TNF受体家族成员9(TNFRSF9)和TRAIL;Bcl-2家族的bcl-2,bcl-w,bax和bak1以及Akt等;表达下调的基因有Fas相关因子(FAF1).结论 糖尿病早期大鼠视网膜血管基因表达发生了复杂的变化.特别是多个凋亡相关通路的基因在糖尿病早期就发生改变,而且多数处在通路上游.提示糖尿病视网膜病变的发生涉及多条凋亡信号传导通路.分子生物化学水平上的变化还仅仅局限在凋亡的诱导期.

abstractsObjective To analyze the expression of apoptosis-relared genes of retinal blood vessel in early diabetic rats by gene chip technology. Methods To make diabetic rat model by intraperitoneal injection of streptozotocin (STZ). On the 6th week after blood pressure increased, 10 rats were executed in Diabetic group and normal control group respectively. 20 retinal blood vessels were extracted and the RNA was isolated. The probe was made of α-32 P-deoxyadenosine triphosphate (dATP)-labeled sample which hybridized 1176 nylon chips, and then analyzed by software. Three different expression genes were selected to verify by reverse transcription polymerase chain reaction (RT-PCR). Results On the 6th week, 136 (11.5%)genes were differentially expressed [up-regulated genes were 90 (7.6%), down- regulated genes were 46(3.9%)] in diabetic group. These genes involved into different groups according to their function. Especially in 72 apoprosis-related genes, 15 genes were differentially expressed. The up- regulated genes were some TNF receptor family members such as TNFRSF12, TRAIL, TNFRSF9, FADD;Bcl-2 family members such as bcl-w, bax, bakl and AKT. The down-regulated genes were FAF1 which related to fas. Conclusions The expression of retinal vascular gene in early diabetic rats has been changed complicatedly. In particular, the multiple apoptosis-related genes have been changed in early diabetic, and most of them are at the upstream of apoptosis pathway. These findings indicate that the development of diabetic retinopathy is associated with multiple signaling pathways leading to apoptosis, while the alterations on the level of molecular biochemistry are still limited in apoptosis induction period.