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目的观察葛根素对肠缺血再灌注大鼠肠黏膜iNOS活性及基因表达的影响。方法♂SD大鼠分为假手术组、肠缺血再灌注2、8 h组、葛根素小、大剂量(70、140 mg.kg-1)+肠缺血再灌注2 h和8 h组、连续ip给药7 d。于第7天给药后30 min麻醉大鼠,复制肠缺血再灌注损伤模型。于再灌注后2、8 h观察大鼠小肠病理形态学的改变并检测血清中一氧化氮(NO)的含量及肠黏膜诱导型一氧化氮合酶(iNOS)的活性,用RT-PCR半定量测定iNOS mRNA的含量。结果大鼠应用葛根素和肠缺血再灌注2 h和8 h后,与模型对照组比较,可减轻肠黏膜的破坏;假手术组大鼠显示较低的黏膜iNOS活性、iNOS mRNA含量及血清NO水平,肠缺血再灌注后则表现出逐渐增加的趋势。至再灌注8 h,模型对照组的iNOS活性、NO水平较假手术组分别增加了64%、75%(P<0.05);而葛根素小、大剂量组则使黏膜的iNOS活性下降46%、50%(P<0.05),血清NO水平下降24%、43%,缺血再灌注2 h,肠黏膜iNOS mRNA表达已明显诱导上调,与再灌注8 h比较无明显差异,葛根素大剂量可使肠黏膜的iNOS mRNA含量显著降低。结论葛根素可降低肠缺血再灌注大鼠肠黏膜的iNOS活性和iNOS mRNA的表达,这可能是葛根素保护肠缺血再灌注损伤的作用机制之一。
Objective To observe the effect of puerarin on iNOS activity and gene expression of intestinal mucosa in intestinal ischemia-reperfusion rats. Methods Sprague-Dawley rats were divided into sham operation group, intestinal ischemia reperfusion group 2 and 8 hours, puerarin small and high dose (70,140 mg.kg-1) + 2 hours and 8 hours after intestinal ischemia reperfusion. Continuous ip administration for 7 d. Rats were anesthetized 30 min after administration on the 7th day to replicate the intestinal ischemia-reperfusion injury model. The pathological changes of the small intestine were observed at 2 and 8 h after reperfusion and the levels of nitric oxide (NO) in serum and inducible nitric oxide synthase (iNOS) in intestinal mucosa were detected. Quantitative determination of iNOS mRNA content. Results Compared with the model control group, rat puerarin and intestinal ischemia reperfusion 2 h and 8 h reduced the damage of the intestinal mucosa; rats in the sham group showed lower mucosal iNOS activity, iNOS mRNA content and serum The level of NO showed a gradually increasing trend after intestinal ischemia-reperfusion. After 8 hours of reperfusion, the iNOS activity and NO level in the model control group were increased by 64% and 75% respectively compared with the sham group (P<0.05), while the small and high doses of puerarin reduced the iNOS activity of the mucosa by 46%. 50% (P<0.05), serum NO levels decreased by 24% and 43%. INOS mRNA expression in the intestinal mucosa was significantly up-regulated after ischemia and reperfusion for 2 h, and there was no significant difference compared with 8 h after reperfusion. Puerarin high-dose Intestinal mucosal iNOS mRNA content can be significantly reduced. Conclusion Puerarin can reduce iNOS activity and iNOS mRNA expression in intestinal mucosa of gut ischemia-reperfusion rats, which may be one of the mechanisms of puerarin protecting intestinal ischemia-reperfusion injury.