硫酸鱼精蛋白联合脂多糖构建间质性膀胱炎动物模型的研究

Protamine sulfate plus lipopolysaccharide in establishment of animal model of interstitial cystitis

目的探讨利用硫酸鱼精蛋白联合脂多糖构建间质性膀胱炎动物模型的可行性。方法将30只雌性SD大鼠分为试验组、环磷酰胺组和对照组,每组各10只。试验组采用10 mg/mL硫酸鱼精蛋白加750μg/mL脂多糖经尿道膀胱灌注;环磷酰胺组给予100 mg/kg的环磷酰胺腹腔内注射;对照组采用磷酸缓冲液膀胱灌注。采用苏木精-伊红染色计数膀胱组织单核炎症细胞,特殊染色计数脱颗粒的肥大细胞;测定各组大鼠尿液中组胺的质量浓度。结果试验组膀胱组织可见大量单核炎症细胞以及较多脱颗粒状态的肥大细胞浸润;环磷酰胺组可见较多单核炎症细胞,但只有少量脱颗粒状态的肥大细胞;对照组只有少量的炎症细胞和未脱颗粒的肥大细胞。试验组和环磷酰胺组每个视野单核炎症细胞计数分别为76.5±9.8和69.3±6.9,均明显高于对照组的16.5±9.8(P<0.05);试验组、环磷酰胺组和对照组每个视野脱颗粒肥大细胞计数分别为8.4±3.1、1.2±1.6和0.7±0.3,试验组明显高于其他两组(P<0.05);试验组、环磷酰胺组和对照组大鼠尿液中组胺的质量浓度分别为(58.5±10.3)、(11.9±2.4)和(8.9±3.2)pg/mL,试验组明显高于其他两组(P<0.05)。结论 10 mg/mL硫酸鱼精蛋白加750μg/mL脂多糖经尿道膀胱灌注构建的间质性膀胱炎动物模型符合间质性膀胱炎的病理生理表现,建模方法简单、可靠。

Objective To investigate the feasibility of establishment of animal model of interstitial cystitis by protamine sulfate plus lipopolysaccharide.Methods Thirty female SD rats were divided into experiment group,cyclophosphamide group and control group,with 10 rats in each group.Transurethral bladder perfusion of 10 mg/mL protamine sulfate plus 750 μg/mL lipopolysaccharide was performed in experiment group,intraperitoneal injection of 100 mg/kg cyclophosphamide was conducted in cyclophosphamide group,and PBS solution perfusion was administered in control group.Mononuclear inflammatory cells in bladder tissues were counted with HE staining,degranulated mast cells were counted with special staining,and the urine mass concentrations of histamine in groups were measured.Results A large number of mononuclear inflammatory cells and degranulated mast cells in bladder tissues were observed in experiment group,a large number of mononuclear inflammatory cells and a small number of degranulated mast cells in bladder tissues were found in cyclophosphamide group,and a small number of mononuclear inflammatory cells and degranulated mast cells in bladder tissues were detected in control group.The numbers of mononuclear inflammatory cells per visual field in experiment group（76.5±9.8） and cyclophosphamide group（69.3±6.9） were significantly larger than that in control group（16.5±9.8）（P0.05）.The number of degranulated mast cells per visual field in experiment group（8.4±3.1） was significantly larger than those in cyclophosphamide group（1.2±1.6） and control group（0.7±0.3）（P0.05）.The urine mass concentration of histamine in experiment group [（58.5±10.3） pg/mL] was significantly higher than those in cyclophosphamide group [（11.9±2.4） pg/mL] and control group [（8.9±3.2） pg/mL]（P0.05）.Conclusion The animal model of interstitial cystitis established by 10 mg/mL protamine sulfate plus 750 μg/mL lipopolysaccharide is in line with the pathophysiological characteristics of interstitial cystitis,and the way of establishment is simple and reliable.