Chen Wei, Fu Xiao-Bing, Ge Shi-Li, Sun Tong-Zhu, Zhou Gang, Han Bing, Du Yi-Ri, Li Hai-Hong, Sheng Zhi-Yong
Wound Healing and Cell Biology Laboratory, 304th Hospital, Burns Institute, Trauma Center of Postgraduate Medical College, 51 Fu Cheng Road, Beijing 100037, China.
World J Gastroenterol. 2005 Jun 14;11(22):3419-25. doi: 10.3748/wjg.v11.i22.3419.
To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanisms of aFGF.
One hundred and eight Wistar rats were randomly divided into sham-operated control group (C) (n = 6), intestinal ischemia group (I) (n = 6), aFGF treatment group (A) (n = 48) and intestinal ischemia-reperfusion group (R) (n = 48). In group I, the animals were killed after 45 min of superior mesenteric artery (SMA) occlusion, while in groups R and A, the rats sustained 45 min of SMA occlusion and were then treated with normal saline and aFGF, respectively, sustained 15 min, 30 min, 1, 2, 6, 12, 24, or 48 h of reperfusion, respectively. In group C, SMA was separated, but without occlusion. Apoptosis in intestinal villus was determined with terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling technique (TUNEL). Intestinal tissue samples were taken not only for detection of bax and bcl-2 gene expression by RT-PCR, but also for detection of bax and bcl-2 protein expression and distribution by immunohistochemical analysis.
The rat survival rates in aFGF treated group were higher than group R (P<0.05) and the improvement of intestinal histological structures was observed at 2, 6, and 12 h after the reperfusion in group A compared with group R. The apoptotic rates were (41.17+/-3.49)%, (42.83+/-5.23)% and (53.33+/-6.92)% at 2, 6 and 12 h after reperfusion, respectively in group A, apparently less than those of group R at matched time points (50.67+/-6.95, 54.17+/-7.86, 64.33+/-6.47, respectively) (P<0.05). The bax gene transcription and translation were significantly decreased in group A vs group R, while mRNA and protein contents of Bcl-2 in group A were obviously higher than those in group R during 2-12 h period after reperfusion.
The changes in histological structure and the increment of apoptotic rate indicated that the intestinal barrier was damaged after intestinal I/R injury, whilst intravenous aFGF could alleviate apoptosis induced by ischemia and reperfusion in rat intestinal tissues, in which genes of bax and bcl-2 might play important roles.
检测酸性成纤维细胞生长因子(aFGF)对大鼠肠缺血/再灌注(I/R)损伤后细胞凋亡及bax和bcl-2基因表达的影响,探讨aFGF的保护机制。
108只Wistar大鼠随机分为假手术对照组(C组)(n = 6)、肠缺血组(I组)(n = 6)、aFGF治疗组(A组)(n = 48)和肠缺血-再灌注组(R组)(n = 48)。I组动物在肠系膜上动脉(SMA)阻断45分钟后处死,而R组和A组大鼠持续SMA阻断45分钟,然后分别用生理盐水和aFGF处理,再分别持续15分钟、30分钟、1小时、2小时、6小时、12小时、24小时或48小时的再灌注。C组分离SMA,但不阻断。用末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记技术(TUNEL)测定肠绒毛中的细胞凋亡。不仅取肠组织样本通过逆转录-聚合酶链反应(RT-PCR)检测bax和bcl-2基因表达,还通过免疫组织化学分析检测bax和bcl-2蛋白表达及分布。
aFGF治疗组大鼠存活率高于R组(P<0.05),且A组在再灌注后2小时、6小时和12小时观察到肠组织结构改善,与R组相比。A组再灌注后2小时、6小时和12小时的凋亡率分别为(41.17±3.49)%、(42.83±5.23)%和(53.33±6.92)%,明显低于R组在相应时间点的凋亡率(分别为50.67±6.95、54.17±7.86、64.33±6.47)(P<0.05)。与R组相比,A组bax基因转录和翻译显著降低;而在再灌注后2 - 12小时期间,A组Bcl-2的mRNA和蛋白含量明显高于R组。
组织结构变化和凋亡率增加表明肠I/R损伤后肠屏障受损,而静脉注射aFGF可减轻大鼠肠组织缺血再灌注诱导的细胞凋亡,其中bax和bcl-2基因可能起重要作用。