中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2008年
4期
244-248
,共5页
糖尿病视网膜病变/病因学%细胞凋亡%寡核苷酸序列分析%糖尿病%实验性
糖尿病視網膜病變/病因學%細胞凋亡%寡覈苷痠序列分析%糖尿病%實驗性
당뇨병시망막병변/병인학%세포조망%과핵감산서렬분석%당뇨병%실험성
Diabetic retinopathy/etiology%Apoptosis%Oligonucleotide array sequenceAnalysis%Diabetes mellitus,experimental
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.
目的 使用基因芯片技術分析糖尿病早期大鼠視網膜血管凋亡相關基因錶達概況.方法 腹腔註射鏈脲佐菌素(STZ)製備糖尿病大鼠模型.在血糖升高後的第6週分彆處死正常組大鼠和糖尿病組大鼠各10隻,提取20隻眼的視網膜血管,一步法提取總RNA.使用(α-32P)脫氧腺苷痠(dATP)標記樣品製作探針,與含有1176箇基因的尼龍膜芯片進行雜交.使用計算機軟件對所穫結果進行相關分析.選擇3箇差異錶達的基因進行逆轉錄聚閤酶鏈反應(RT-PCR)驗證.結果 糖尿病大鼠第6週,136箇基因具有差異錶達.佔檢測基因總數的11.5%.其中.錶現為上調的基因90箇,佔7.6%;錶現為下調的基因46箇,佔3.9%.差異錶達涉及多種功能的多箇基因.與凋亡信號傳導通路相關的72箇基因中.有15箇齣現瞭錶達的差異.錶達上調的基因包括腫瘤壞死因子(TNF)傢族中Fas相關的死亡域(FADD)、TNF受體傢族成員12(TNFRSF12)、TNF受體傢族成員9(TNFRSF9)和TRAIL;Bcl-2傢族的bcl-2,bcl-w,bax和bak1以及Akt等;錶達下調的基因有Fas相關因子(FAF1).結論 糖尿病早期大鼠視網膜血管基因錶達髮生瞭複雜的變化.特彆是多箇凋亡相關通路的基因在糖尿病早期就髮生改變,而且多數處在通路上遊.提示糖尿病視網膜病變的髮生涉及多條凋亡信號傳導通路.分子生物化學水平上的變化還僅僅跼限在凋亡的誘導期.
목적 사용기인심편기술분석당뇨병조기대서시망막혈관조망상관기인표체개황.방법 복강주사련뇨좌균소(STZ)제비당뇨병대서모형.재혈당승고후적제6주분별처사정상조대서화당뇨병조대서각10지,제취20지안적시망막혈관,일보법제취총RNA.사용(α-32P)탈양선감산(dATP)표기양품제작탐침,여함유1176개기인적니룡막심편진행잡교.사용계산궤연건대소획결과진행상관분석.선택3개차이표체적기인진행역전록취합매련반응(RT-PCR)험증.결과 당뇨병대서제6주,136개기인구유차이표체.점검측기인총수적11.5%.기중.표현위상조적기인90개,점7.6%;표현위하조적기인46개,점3.9%.차이표체섭급다충공능적다개기인.여조망신호전도통로상관적72개기인중.유15개출현료표체적차이.표체상조적기인포괄종류배사인자(TNF)가족중Fas상관적사망역(FADD)、TNF수체가족성원12(TNFRSF12)、TNF수체가족성원9(TNFRSF9)화TRAIL;Bcl-2가족적bcl-2,bcl-w,bax화bak1이급Akt등;표체하조적기인유Fas상관인자(FAF1).결론 당뇨병조기대서시망막혈관기인표체발생료복잡적변화.특별시다개조망상관통로적기인재당뇨병조기취발생개변,이차다수처재통로상유.제시당뇨병시망막병변적발생섭급다조조망신호전도통로.분자생물화학수평상적변화환부부국한재조망적유도기.
Objective 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.
