脓毒症大鼠心肌细胞凋亡与p53和Bcl-2基因表达的关系  

Correlation between the myocardial apoptosis and the expression of the genes p53 and bd-2 in septic rats

作　　者：梁艳冰[1] 唐皓[1] 翟诚顺 陈志斌[1] 李振宇[1] 何东华[2] 马中富[1] 

机构地区：[1]中山大学附属第一医院普内科,广州510080 [2]中山大学达安基因公司 [3]深圳市武警总队医院急诊科

出　　处：《中华急诊医学杂志》2008年第2期128-131,共4页Chinese Journal of Emergency Medicine

基　　金：广东省科技计划项目（2004830601002）;广州市科委重点课题基金（2001-Z-130-02）

摘　　要：目的探讨脓毒症大鼠心肌细胞凋亡的变化及其与Bcl-2和茚3基因mRNA表达的关系。方法取体重190—220g的成年雄性sD大鼠42只，随机分为三组，脓毒症组30只，正常对照组6只，假手术组6只；脓毒症组以盲肠结扎穿刺法（CLP）复制脓毒症大鼠模型，于术后3h、9h、12h、24h各处死6只大鼠，假手术组麻醉满意后打开腹腔，正常对照组仅行麻醉术，均取1．0mm^3大小的心肌组织，另外余下的6只脓毒症鼠术后待其自然死亡不取心肌。采用电镜和TUNEL法检测大鼠心肌细胞的凋亡，用RT—PCR方法检测大鼠心肌细胞Bcl-2和p53基因mRNA的表达。结果电镜下大鼠凋亡的心肌细胞胞膜皱缩，核固缩、边集、碎裂，胞膜突起、凋亡小体形成，TUNEL法检测各时间点的脓毒症大鼠心肌细胞的凋亡率均较正常对照组和假手术对照组明显升高（P〈0．05），术后12h达到峰值；p53mRNA表达明显升高（P〈0．05），其表达量与心肌细胞凋亡指数呈正相关性（r=0．976，P〈0．05）；Bcl-2 mRNA表达明显降低（P〈0．05），其表达量与心肌细胞凋亡指数呈负相关性（r=-0．833，P〈0．05）。结论脓毒症大鼠心肌细胞的凋亡明显增高，p53和Bcl-2 mRNA的表达异常是其可能机制之一。Objective To study the myocardial apoptosis and the correlation between the apoptosis and the mRNA expression of the genes p53 and Bcl-2 in septic rats. Method Forty-two male SD rats （weight 190 - 220 g） were randomly divided into three groups, namely, sepsis group （ n = 30）, normal control group （ n = 6）, and sham-operated control group （ n = 6）. The septic animal model were reproduced by cecal ptmcture and ligation. After operations, 6 septic rats were sacrificed at various intervals 6 h, 9 h, 12 h and 24 h. In the sham- operated control group the rats＇ abdominal cavity was opened after effective anaesthesia. The normal control rats were just taken anaesthesia. Then approximately 1.0 mm3 of their myocardial tissues were taken respectively. The remaining 6 septic rats died naturally after operation without being taken myocardial tissues. The myocardial apoptosis change in rats were detected by electromicroscope and TdT-mediated dUTP Nick End Labeling （TUNEL）. The mRNA expression of genes p53 and Bcl-2 were measured by reverse transcriptional polymerase chain reaction （rt-PCR）. Results Under electromieroseope the myocardial apoptosis in rats was showed as cell membrane shrinkage, katyopyenosis, side collection and fragmentation of nuclear, formation of membrane prominence and apoptotie body. At each time points the myocardial apoptosis of septic rats measured by TUNEL were significant higher than that of normal and sham-operated control group （ P 〈 0.05 ）, while the climax was presented at 12 hr after the operations. And the mRNA expression of p53 gene in septic rats were significant higher than that of normal and sham-operated control group （ P 〈 0.05 ）. There was positive correlation between the mRNA expression of p53 gene and the myocardial apoptotie index （ r = 0.976, P 〈 0.05）. However, the mRNA expression of Bcl-2 gene was significant lower than that of normal control group and sham-operated control group （P 〈 0.05）, and there was negative correlation betwe

关 键 词：脓毒症 细胞凋亡 心肌损害 P53 BCL-2 

分 类 号：R285.5[医药卫生—中药学]