不同剂量熊果酸对大鼠静脉桥再狭窄过程中细胞凋亡的影响

Study of ursolic acid induce vein cells apoptosis in rats vein graft restenosis model with different doses of urslic acid

目的 研究不同剂量的熊果酸诱导大鼠静脉桥再狭窄过程中内膜和中膜细胞凋亡的作用.方法 SD大鼠60只,体重300~350 g,随机分成UA（熊果酸）0组、UA1组、UA2组和UA3组.将所有实验动物的右侧颈外静脉以套管法移植至右侧颈总动脉上.UA1组、UA2组和UA3组术后首日开始予以熊果酸灌胃,剂量分别为15 mg·kg-1·d-1、30 mg·kg-1·d-1和45 mg·kg-1·d-1,UA0组不予熊果酸干预.术后满第1周、第2周和第4周分三次取材,每次每组随机选取5只大鼠.每条静脉标本随机选取不同位置的切片共5张做TUNEL检测.计算凋亡细胞指数.将4组数据对比研究.结果 1周、2周、4周静脉桥内膜细胞凋亡指数：UA0组（22.0±4.1）%、（18.6±2.8）%、（8.4±3.6）%,UA1组（28.4±2.4）%、（19.2±1.4）%、（9.4±3.5）%,UA2组（35.2±1.7）%、（25.6±2.7）%、（11.6±2.7）%,UA3组（41.5±1.9）%、（28.4±1.8）%、（16.3±1.4）%.1周、2周、4周静脉桥中膜细胞凋亡指数：UA0组（4.0±2.2）%、（2.6±1.5）%、（2.0±0.7）%,UA1组（6.6±2.3）%、（4.2±1.6）%、（2.6±1.1）%,UA2组（7.8±3.7）%、（4.4±2.1）%、（3.4±1.1）%,UA3组（7.6±1.7）%、（4.0±0.8）%、（3.5±0.7）%.内膜细胞凋亡指数比较,时间效应的P〈0.05,组别效应的P〈0.05,交互效应的P〈0.05.中膜细胞凋亡指数比较,时间效应的P〈0.05,组别效应的P〉0.05,交互效应的P〉0.05.结论 熊果酸具有诱导大鼠静脉再狭窄的内膜细胞凋亡的作用,该作用随着熊果酸剂量的增大而加强;而熊果酸不具有诱导静脉桥中膜细胞凋亡的作用.

Objective To study the induction of vein intimal and media cells apoptosis in rats vein graft restenosis model with different doses of ursolic acid. Methods 60 SD rats,weighing 300-350 g,were randomly divided into group UA0,group UA1,group UA2 and group UA3. The right external jugular veins were grafted into the right carotid artery with cuff technique for 60 rats. Ursolic acid 15 mg·kg-1·d-1 was administrated daily in rats of group UA1. Ursolic acid 30 mg·kg-1·d-1 was administrated daily in rats of group UA2. Ursolic acid 45 mg·kg-1·d-1 was administrated daily in rats of group UA3. Group UA0 not ursolic acid intervention. Samples of veins tissues were harvested at 1,2,4 weeks. 5 rats were randomly selected each time and each group. 5 slices at different po-sitions were randomly selected in each vein samples. Apoptotic index were calculated after TUNEL test. Apoptotic index of 4 groups was compared. Results Apoptotic index of intimal cells of one week,two weeks and four weeks：group UA0 （22.0 ±4.1）%,（18.6 ±2.8）%,（8.4 ±3.6）%,group UA1 （28.4 ±2.4）%,（19.2 ±1.4）%,（9.4 ± 3.5）%,group UA2 （35.2±1.7）%,（25.6±2.7）%,（11.6±2.7）%,group UA3 （41.5±1.9）%,（28.4±1.8）%,（16.3± 1.4）%. Apoptotic index of media cells of one week,two weeks and four weeks：group UA0 （4.0 ±2.2）%,（2.6 ± 1.5）%,（2.0±0.7）%,group UA1 （6.6±2.3）%,（4.2±1.6）%,（2.6±1.1）%,group UA2 （7.8±3.7）%,（4.4±2.1）%, （3.4±1.1）%,group UA3 （7.6±1.7）%,（4.0±0.8）%,（3.5±0.7）%. Comparison of apoptotic index of intimal cells, the time effect was P〈0.05,the group effect was P〈0.05,the interaction effect was P〈0.05. Comparison of apop-totic index of media cells,the time effect was P〈0.05,the group effect was P〉0.05,the interaction effect was P〉0.05. Conclusion Ursolic acid has the effect of inducing intimal cells apoptosis in rats with venous restenosis. This effect enhanced with increasing ursolic acid dose. While ursolic acid has not the effect of inducing media cells apoptosis.