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目的:探讨葛根素注射液对肠缺血再灌注大鼠肠粘膜细胞凋亡的影响。方法:♂SD大鼠分为5组:假手术组、肠缺血再灌注2h、8h组,葛根素+肠缺血再灌注2h、8h组,每组10只,连续ip给药7d,于d7给药后30min麻醉,复制肠缺血再灌注模型。于再灌注后2h、8h观察小肠病理形态学改变并检测肠粘膜GSH、MDA含量,用脱氧核糖核苷酸末端转移酶介导的末端标记法TUNEL检测肠粘膜细胞凋亡的阳性细胞数,采用免疫组织化学SP法测定半胱氨酸天门冬氨酸特异蛋白酶Caspase-3的表达水平。结果:肠缺血再灌注可使肠粘膜上皮细胞的凋亡数明显增加,且再灌注2h显著高于8h,Caspase-3的表达水平在模型对照组显著升高,再灌注2h与8h无明显差异。葛根素组可使肠粘膜GSH显著升高,MDA显著降低,TUNEL染色阳性细胞数及Caspase-3平均灰度值均显著低于对照组。结论:葛根素可增强机体抗氧化功能、清除有害自由基,并可能通过抑制Caspase-3的表达而抑制肠粘膜细胞调亡。
Objective: To investigate the effect of puerarin injection on intestinal mucosal cell apoptosis after intestinal ischemia-reperfusion in rats. METHODS: Sprague-Dawley rats were divided into five groups: sham operation group, intestinal ischemia reperfusion group 2h and 8h, puerarin+intestinal ischemia reperfusion 2h and 8h groups, 10 in each group, 7 days after continuous ip administration. Anesthesia was performed 30 minutes after administration of d7, and the intestinal ischemia reperfusion model was replicated. At 2 and 8 hours after reperfusion, the pathological changes of the small intestine were observed and the contents of GSH and MDA in the intestinal mucosa were measured. The number of positive cells in the intestinal mucosal cells was detected by terminal deoxynucleotidyl transferase mediated TUNEL assay. Immunohistochemical SP method was used to determine the expression of caspase-3, a caspase-specific protease. Results: The number of apoptotic cells in intestinal epithelium increased significantly after intestinal ischemia-reperfusion, and it was significantly higher than that at 8h after reperfusion for 2h. The expression level of Caspase-3 was significantly increased in the model control group, and there was no obvious reperfusion at 2h and 8h. difference. Puerarin significantly increased GSH in intestinal mucosa and decreased MDA significantly, and the number of TUNEL positive cells and Caspase-3 average gray value were significantly lower than those in control group. Conclusion: Puerarin can enhance the body’s anti-oxidation function, eliminate harmful free radicals, and may inhibit the apoptosis of intestinal mucosal cells by inhibiting the expression of Caspase-3.