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目的:通过对膀胱感染性结石大鼠膀胱灌注脂质体包裹的人β-防御素-2重组真核表达载体(pCAGG-hBD2),观察人β-防御素-2(hBD2)基因对感染性结石的预防作用。方法:选择健康成年雄性SD大鼠40只,随机分为实验组与对照组。实验组膀胱灌注pCAGG-hBD2 250μl,对照组经同法给予等量空载体(pCAGG)。2天后,两组均随机处死3只大鼠,利用小动物成像仪观察重组hBD2在SD大鼠膀胱黏膜的表达;转染成功后,两组均膀胱内置入含菌(奇异变形杆菌)聚乙烯管异物,分别于24、48、72小时收集尿液,进行尿液菌落计数、白细胞计数。30天处死全部大鼠获取膀胱组织,观察结石的形成情况及膀胱组织病理学变化。结果:实验组大鼠置入含菌(奇异变形杆菌)聚乙烯管异物24、48、72小时后,尿液中奇异变形杆菌菌落总数、尿白细胞计数均显著低于对照组,组间比较差异均有统计学意义(P<0.001);实验组和对照组不同时间大鼠尿液中奇异变形杆菌菌落总数及尿白细胞计数差异有统计学意义。对照组大鼠的尿路组织主要表现为肾输尿管膀胱的炎症改变,膀胱内有感染性结石的形成;实验组大鼠的尿路组织病理学无明显变化,膀胱内无结石生成。结论:重组hBD2基因对奇异变形杆菌致泌尿系感染性结石有预防作用。
OBJECTIVE: To investigate the effect of human beta-defensin-2 (hBD2) gene on the infectivity of bladder infectious bladder rat by intraperitoneal perfusion of liposome-encapsulated human β-defensin-2 recombinant plasmid pCAGG-hBD2 Prevention of stones. Methods: Forty adult healthy male SD rats were randomly divided into experimental group and control group. The experimental group was intraperitoneally infused with pCAGG-hBD2 250μl, and the control group was given the same amount of empty vector (pCAGG) by the same method. Two days later, three rats were randomly sacrificed in both groups, and the expression of recombinant hBD2 in bladder mucosa of SD rats was observed by using a small animal imager. After successful transfection, both groups were filled with bacteria containing bacteria (Proteus mirabilis) Tube foreign body, respectively, 24,48,72 hours to collect urine, urine colony count, white blood cell count. After 30 days, all the rats were sacrificed and the bladder tissue was obtained to observe the formation of stone and histopathological changes of bladder. Results: After 24, 48 and 72 hours, the total number of bacteria and urine leukocyte in the urine of experimental group were significantly lower than that of the control group (P <0.001). The total number of urinary albumin and urine leucocyte in the urine of experimental group and control group were significantly different at different time points. In the control group, the urinary tract tissues mainly showed the changes of inflammation in the ureter and bladder, and the formation of infectious stones in the bladder. The pathology of the urinary tract in the experimental group showed no obvious changes and no stones were formed in the bladder. Conclusion: The recombinant hBD2 gene can prevent the urinary tract infection caused by Proteus mirabilis.