改构型酸性成纤维细胞生长因子对肠缺血-再灌注的保护作用
孙同柱 付小兵 陈伟 郑曙云 翁立新 赵京禹 黎君友 孙丹【摘要】 目的 观察改构型酸性成纤维细胞生长因子(rhaFGF)对大鼠肠缺血-再灌注所致肠道损伤的保护作用。方法用夹闭肠系膜上动脉(SMA)造成肠缺血45 min、松夹形成缺血-再灌注的方法制备肠缺血-再灌注损伤模型。生理盐水对照组和rhaFGF治疗组于再灌注即刻分别经静脉注射生理盐水0.1 ml和rhaFGF 4 μg。缺血-再灌注后2、6、12和24 h取血及小肠组织，分别测定D-乳酸含量，观察小肠组织学改变；用末端脱氧核糖转移酶介导的生物素化脱氧尿嘧啶缺刻标记技术(TUNEL)检测肠道细胞凋亡率，并与假手术组比较。结果缺血-再灌注6 h生理盐水对照组D-乳酸含量较假手术组明显升高，达到(0.34±0.09)mg/L，rhaFGF治疗组为(0.23±0.07)mg/L(P均<0.05)。组织学检查显示，缺血-再灌注2~24 h生理盐水对照组肠道损伤严重，rhaFGF治疗组肠道损伤有所减轻。细胞凋亡检测显示，假手术组肠道细胞凋亡率很低；随着缺血-再灌注损伤时间的延长，生理盐水对照组和rhaFGF治疗组细胞凋亡率均明显升高，于缺血-再灌注12 h达峰值(生理盐水对照组细胞凋亡率高达(62.8±1.7)%，而rhaFGF治疗组为(42.5±2.6)%)后均呈下降趋势，两者各时间点均有统计学差异(P均<0.05)。结论 rhaFGF能降低大鼠肠缺血-再灌注所致的血浆D-乳酸含量和肠道细胞凋亡率，提示对肠道损伤有一定的保护作用。
【关键词】 酸性成纤维细胞生长因子； 缺血-再灌注损伤； 小肠； D-乳酸； 细胞凋亡
Protective effects of modified reconbinant human acidic fibroblast growth factor on small intestina after ischemia/reperfusion injury in rats
SUN Tong-zhu, FU Xiao-bing, CHEN Wei, ZHENG Shu-yun, WENG Li-xin, ZHAO Jing-yu, LI Jun-you, SUN Dan.
Key Laboratory of Wound Repair, 304 th Clinical Department of General Hospital of PLA, Beijing 100037， China
【Abstract】 Objective To explore the protective effect of modified recombinant human acidic fibroblast growth factor (rhaFGF) on small intestinal after ischemia/reperfusion(I/R) injury in rats. Methods The clamp on the superior mesenteric artery (SMA) was removed after clamping it for 45 minutes to replicate I/R injury of the intestine in the rat. Rats were then divided randomly into sham operation group, normal saline treatment group and rhaFGF treatment group, in which the rats of the normal saline treatment group were injected 0.1 ml of normal saline and that of the rhaFGF group were given 4 ug of rhaFGF immediately after reperfusion. The content of D-lactate in the plasma was determined and the changes in intestinal pathology were observed. At the same time, the rates of apoptosis of intestinal epithelial cells were assessed 2, 6, 12 and 24 hours after I/R, and compared to the sham operation group. Results The plasma content of D-lactate in the saline treatment group at 6 hours after I/R reached (0.34±0.09)mg/L and was significantly higher than that in the rhaFGF treatment group((0.23±0.07)mg/L,P<0.05). It was shown histologically that the intestinal structures were damaged more seriously in saline treatment group than in rhaFGF group. The apoptosis rates in the saline treatment group and rhaFGF group were elevated significantly, peaking at 12 hours after I/R injury((62.8±1.7)% in saline group and (42.5±2.6)% in rhaFGF treatment group), then the rate began to fall. There was statistically significant differone between the two groups at 12 hours after I/R injury. Conclusion rhaFGF can reduce content of D-lactate in the plasma and rate of apoptosis of epithelial cells in the intestine after I/R injury, thus it seems to afford a protective effect on the small intestine after I/R injury in rats. ......
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