人β-防御素-2基因治疗大鼠泌尿系感染的实验研究  

Anti-infection effect of human beta-defensin-2 gene therapy in a rat urinary tract infection model

作　　者：赵俊丽[1] 王俭勤[1] 王志平[2] 

机构地区：[1]兰州大学第二医院肾病科,730030 [2]兰州大学第二医院泌尿研究所,730030

出　　处：《中华泌尿外科杂志》2011年第12期846-849,共4页Chinese Journal of Urology

基　　金：甘肃省自然科学基金项目（0803RJZA007）;兰州大学中央高校基本科研业务专项资金（lzujbky2010-149）

摘　　要：目的观察基因重组人β-防御素-2（human beta—defensin-2，hBD2）对大鼠模型大肠埃希菌致泌尿系感染的保护作用。方法健康成年雌性SPF级Wistar大鼠56只，随机分为hBD，组与对照组，hBD2组大鼠经逆行膀胱灌注给予脂质体包裹的hBD2重组真核表达载体（pCAGG—hBD2）250μl，对照组同法给予等量空载体（pCAGG），48h后2组大鼠均膀胱灌注1×10^8CFU／mlUT189200μl，制备大肠埃希菌泌尿系感染大鼠模型，分别于不同时间点处死大鼠获取膀胱组织及尿液进行菌落计数、尿液白细胞计数，观察膀胱组织病理学变化并进行炎症评分。结果在感染后24、48、72h，hBD2组大鼠尿液UT189菌落计数分别为（6．43±1．04）、（5．66±0．68）、（1．55±1．17）Log10CFU／ml；膀胱组织UT189菌落计数分别为（6．07±0．52）、（5．17±1．21）、（1．18±0．84）Log10CFU／g；尿白细胞计数分别为（25．1±2．9）、（16．4±7．1）、（11．0±3．0）×10^4cells／ml；膀胱组织炎症评分（2．21±0．45）、（1．30±0．40）、（1．28±0．28）。对照组大鼠尿液UT189菌落计数分别为（8．85±0．79）、（8．07±1．30）、（7．93±2．89）Log10CFU／ml；膀胱组织UT189菌落计数分别为（7．82±1．29）、（7．99±1．71）、（6．73±1．59）Log10CFU／g；尿白细胞计数分别为（168．0±14．9）、（234．5±31．3）、（270．0±22．1）×10^4cells／ml；膀胱组织炎症评分（3．30±0．41）、（4．13±0．13）、（4．72±0．24）。hBD2组大鼠感染后24，48和72h尿液及膀胱组织中大肠杆菌菌落计数、尿白细胞计数及组织炎症评分均显著低于对照组，组间差异均有统计学意义（P〈0．05）。结论基因重组hBD2对大肠埃希菌致泌尿系感染有显著的预防及治疗作用，可能与其杀菌作用和抑制炎症反应有关。Objective To assess the human β-defensin-2 （hBD2） gene therapeutic efficacy in a rat urinary tract infection （UTI） model via intravesical liposome-mediated gene transfer. Methods Fifty-six female Wistar rats （class SPF） weighting 180 -220 were randomly divided into an experiment group and a control group （each n = 28）. The animals were administered either 250 μl recombinant pCAGG-hBD2 or control vector pCAGG intravesically. After 48 h, rats in both groups were infected via intravesical inoculation with 200μl of the bacterial suspension of UTI89 （ 1 ×10^8 CFU/ml）. The rats were sacrificed at 4, 24, 48, and 72 h post-inoculation. The bladders were aseptically removed and bisected. One half was fixed in neutral buffered formalin for histological analysis. The remaining half-bladders were homogenized and titered for surviving bacteria. The clean-catch urine sample from each rat was collected in sterile before they were killed for bacterial titers determined and WBC counted. Results Numbers of bacterial colony-forming unit in urine and bladders from hBD2 gene treated UTI rats were significantly lower than those from the control vectoradministered UTI rats at 24, 36, and 72 h after infection （ P 〈 0.05 ）. The amount of WBC in urine was significantly less in the defensin group than in the control group. In addition, in vivo expression of hBD2 could reduce mucosa damage, interstitium edema and inflammatory cells infiltration in UTI animals. Conclusions The successful inhibition of UTI progression could be obtained with hBD2 gene therapy. Recombinant beta-defensin-2 could kill UTI89 in vivo and suppress the subsequent infection-induced inflammatory responses.

关 键 词：人Β-防御素-2 基因治疗 泌尿系感染 

分 类 号：R691.3[医药卫生—泌尿科学]