高脂饮食大鼠血清抵抗素水平及下丘脑细胞因子信号转导抑制因子_3基因表达的变化(1)

脑组织 100~ 120 m g, 用 T rizo l一步法提取总 mRNA, 经电泳显示 28、18、5 S清晰条带, 测定 A 260 /A280 > 1. 8。应用反转录聚合酶链反
应 ( RT PCR )检测其 SOCS 3 m RNA 表达水平。 PCR产物经 18 g /L 琼脂糖凝胶电泳后, 利用凝胶图像分析系统测定电泳条带密度
( 1. 南京医科大学附属南京妇幼保健院 儿童保健科, 南京 210004; 2. 中国人民解放军第 82医院 内分泌科, 江苏 淮安 223001; 3. 南京医科大学 儿科医学研究所, 南京 210029 )
摘要: 目的 观察高脂饮食大ቤተ መጻሕፍቲ ባይዱ血清抵抗素水平及下丘脑细胞因子信号转导抑制因子 3 ( SOCS 3)基因表达变化及其关系, 探
A bstrac t: Objective T o observe the concentrations of serum resistin and changes o f suppressor o f cytokine signa ling - 3( SOCS- 3) gene expression in hypothalam us o f rats fed w ith high fat die,t and to exp lore the pathogenesis of obesity. M ethods The m ale ra ts aged 6 weeks, w eighted 80- 100 g were allocated into 2 groups random ly, each containing 8 rats. The rats of h igh fat diet group were fed w ith h igh fat die,t the rats of basic diet group were fed w ith basic die.t The ra ts were housed in cages indiv idually at a tem perature of 21- 23 , m aintained on a light- dark cycles of 12 hours, given free access to tapw ater and food. Body weigh,t body leng th, and tail leng th were recorded every week. A fter 9 weeks, the blood in heart of ratswas collected during the depth of urethane ( 0. 4 g /kg) anesthesia for the fo llow ing research. The concen tration of serum resistin and glucose were m easured by enzym e- linked mi m unosorbent assay and glucose ox idase m ethod respectively. The hypo thalamus w ith 100- 120 m g was rem oved, ofwhich the total RNA w as collected by using T rizol reagen,t 28 S, 18 S and 5 S bandsw ere a ll c lear in electrophoresis picture and the ratio A260 /A280 w asm ore than 1. 8. The level o fSOC S- 3 gene mRNA expressionw as evaluated by reverse tran scr iption polym erase cha in reaction ( RT - PCR). The PCR productsw ere exam ined by electrophoresis w ith 1. 8% agarose ge l and observed by gelmi age ana lysis system. The relative am ount o fSOCS- 3mRNA wasm easured norm alizing to the mRNA levels of g lyceraldchyde 3- phospha te dehydrogenase ( GAPDH ) and expressed as the percentage o f density of SOCS- 3 band w ith density of GAPDH band. Statistica l ana lysis was perform ed using the SPSS 15 0 statistical software program. R esults 1. The w eigh,t body length, ta il leng th and L ee s index of rats fed w ith high fat diet increased sign ificantly than those of basic diet group(Pa < 0. 001). 2. The rats fed w ith h igh fat diet showed of significantly increased serum glucose and resistin levels com paredw ith those o f ra ts fed w ith basic diet(Pa < 0. 01). 3. The levels of SOCS- 3 genemRNA expression in hypothalam us of rats fed w ith h igh fat dietwere h igher than those of rats fedw ith basic diet (P < 0. 05). 4. The serum resistin levels of 2 groups had no corre lation w ith the leve ls of SOCS- 3 mRNA (Pa > 0. 05). C onclusions H igh fa t diet can induce the serum resistin levels of rats an in crease, and in turn results in elevated expression of SOCS- 3 gene. The induction of SOCS- 3 by resistinm ay be a m ediator in the process of in sulin resistance induced by obesity and resistin.
值, 以甘油醛 3磷酸脱氢酶 ( GAPDH )为内参照, 计算 SOCS 3m RNA 相对水平, 结果以 SOCS 3条带占 GAPDH 条带密度百分率 (% )
表示。结果 1. 高脂饮食组大鼠体质量、体长、尾长及 L ee s指数明显高于基础饮食组 (P a < 0. 001); 2. 高脂饮食组大鼠血清血糖及 抵抗素水平明显升高, 与基础饮食组比较均有显著性差异 (P a < 0. 01); 3. 高脂饮食组大鼠下丘脑 SOCS 3 mRNA 表达水平显著高于
调节因子。抵抗 素是 S teppan 等 [ 1] 发现 的一 个新 的脂 肪 因子, 与肥胖 及胰 岛素 抵抗 的发 生发 展密 切相 关 [ 2 4], 但 其确切机制目 前尚 不清 楚。研究 报道, 抵抗 素具 有诱 导 3T3L 1 脂 肪 细 胞 表 达 细 胞 因 子 信 号 转 导 抑 制 因 子 3 ( suppressor of cytok in e signaling - 3, SOCS 3 ) 的 作 用 [ 5] ,
J Appl C lin Ped iatr, 2008, 23 ( 20): 1573- 1575 K ey words: resistin; suppressor of cy tokine signa ling- 3; obesity; high- fat die;t ra t
脂肪组织分泌的许多化学 物质是机体糖 稳态的重 要
基础饮食组 (P < 0. 05); 4. 二组大鼠血清抵抗素水平与下丘脑 SOCS 3 mRNA 表达水平无明显相关性 (P a > 0. 05)。结论 高脂饮食
可导致大鼠血清抵抗素水平升高, 进而诱导 SOCS 3基因表达, 在肥胖、抵抗素所致的胰岛素抵抗过程中 SOCS 3可能发挥了重要的
介质作用。
讨高脂饮食引发肥胖的发生机制。方法 雄性 6周龄 SD 大鼠 16只。体质量 80~ 100 g。随机分为基础饮食组和高脂饮食组, 每组
8只。单笼饲养, 自由饮食饮水, 明暗周期 12 h, 环境温度 21~ 23 , 每周称体质量、测体长和尾长 1次。大鼠饲养 9周后, 以乌拉坦
( 0 4 g /kg)深度麻醉, 取心脏全血离心, 采用 EL ISA 法检测其血清抵抗素水平, 葡萄糖氧化酶法检测其血清葡萄糖水平。取其下丘
实用儿科临床杂志, 2008, 23 ( 20): 1573- 1575
关键词: 抵抗素; 细胞因子信号转导抑制因子 3; 肥胖症; 高脂饮食; 大鼠
中图分类号: R 725. 8
文献标志码: A
文章编号: 1003- 515X( 2008) 20- 1573- 03
Changes of Serum Resistin L evels and Suppresso r of Cytok ine Signa ling- 3 G ene Expression in H ypo tha lamus