摘要
目的 研究四逆汤(SND)对大鼠肠缺血再灌注 (II/R后肠粘膜细胞凋亡的影响,并从神经酰胺信使通路探讨其抗凋亡机制。方法 24只SD大鼠随机分为 3组(n=8):对照组(仅分离不阻断肠系膜上动脉)、模型组 (阻断肠系膜上动脉 1h后再灌注 3h)、SND组(每天 3g·kg-1 SND灌胃,连续 3天后手术)。取回肠末端组织行电镜检查;检测肠粘膜组织SOD活性、MDA及神经酰胺含量的变化,TUNEL法检测肠粘膜上皮细胞的凋亡指数,RT PCR法分析肠粘膜组织鞘磷脂酶(SMase)基因表达的变化。结果 模型组电镜下可发现许多典型凋亡的上皮细胞,凋亡指数达 30 82%6 34%,SND预处理后凋亡指数为 14 91%±5 40%,明显低于模型组(P<0 01);模型组SOD活性明显低于对照组 (<0 01),MDA与神经酰胺含量以及SMase的基因表达明显高于对照组(P<0 01),且神经酰胺含量与凋亡指数具有良好的正相关 (r=0 852,P<0 01),与SOD活性具有负相关(r= -0 775,P<0 01 )。SND预处理能明显增强SOD活性,降低肠粘膜组织MDA及神经酰胺含量,减少肠粘膜组织SMase的基因表达(P<0 01)。SND组凋亡指数与神经酰胺的含量亦呈显著的正相关 (r=0 832,P<0 01 )。结论 SND具有抗II/R后肠粘膜细胞凋亡的作用,其作用与它清楚氧自由基、抑制SMase的基因表达、减少神经酰胺的生成有关。
Aim To investigate the effects of sini decoction(SND) on intestinal mucosa cell apoptosis and its mechanism after intestinal ischemia reperfusion in rats.Methods Twenty-four Sprague-Dawley rats of both sexes were randomly divided into 3 equal groups:①control group in which sham operation was performed;②model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours;③SND group in which SND(3 g·kg -1)was given via stomach tube for 3 days before operation. A strip of small intestine was taken from distal end of ileum for electron microscopic examination; SOD activity, MDA and ceramide concentration of intestinal mucosa were detected;Apoptosis of intestinal mucosa cell was examined by TUNEL method and Sphingomyelinase (SMase) mRNA expression was analyzed by RT-PCR.Results In model group, many typical apoptotic epithelial cells were seen under electron microscopic, apoptosis index was higher than that in control group(P<0.01). SOD activity of model group was significantly lower than that of control group(P<0.01), but MDA and ceramide content as well as SMase mRNA expression were obviously higher than those in control group(P<0.01).In addition, In model group, ceramide content was positively correlated to apoptosis index(r=0.852,P<0.01),and negativel y to SOD activity (r=-0.775,P< 0.01).The pretreatment of SND change d the above abnormal phenomena significantly(P<0.01). Conclusion SND can reduce intestinal mucosa cell apoptosis by removing oxygen free r adical, downregulation of SMase mRNA expression and suppression of ceramide production.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2005年第2期240-244,共5页
Chinese Pharmacological Bulletin
基金
广东省中医药管理局基金资助项目(No1040066)
关键词
四逆汤
肠粘膜
再灌注损伤
凋亡
神经酰胺类
鞘磷脂酶
sini decoction
intestinal mucosa
reperfusion injury
apoptosis
ceramide
sphingomyelinase
