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摘要: 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 250 μl,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。Abstract: Objective:To observe therapeutic efficacy of human β-defensin-2 (hBD2) gene on infection stones caused by proteus mirabilis in rat models.Method:Forty Sprague-Dawlay rats were randomly divided into an experimental group and a control group. The experimental group were administered 250 μl recombinant pCAGG-hBD2 and control group were administered vector pCAGG intravesically respectively. Two days later, three rats of both groups were randomly sacrificed respectively in order to assay the expression of recombinant hBD2 in rats of bladder mucosa by small animal imaging. Then, rats of both groups were infected via proteus mirabilis inoculation with a polyethylene pipe, which had been contaminated by the proteus mirabilis. At 24, 48 and 72 h post-inoculation we collected urine samples from each rat for bacterial titers determined and WBC counted. All rats were sacrificed one month later, the bladders were aseptically removed and bisected for observing the formation of stones and histological analysis.Result:Numbers of bacterial colony-forming unit in urine from hBD2 gene treated urinary tract infection stone in experimental rats were significantly lower than those from the control vector administered rats at 24, 48, and 72 h after infection (P<0.001). The amount of WBC in urine was significantly less in experimental group than in the control group. In addition, the histopathologic changes were mainly inflammatory changes for kidney, ureter and bladder in control group. Infection stones formation in bladder was found in control group. No changes or stone formation in kidney, ureter and bladder were found in experimental group.Conclusion:The successful inhibition of urinary tract infection stone formation progression could be obtained with hBD2 gene therapy.
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Key words:
- infection stone /
- human β-defensin-2 /
- genetic prevention
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