Establishment of a Novel Autoimmune Experimental Model of Bladder Pain Syndrome/Interstitial Cystitis in C57BL/6 Mice
Establishment of a Novel Autoimmune Experimental Model of Bladder Pain Syndrome/Interstitial Cystitis in C57BL/6 Mice
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DOI：
10.1007/s10753-017-0531-7
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发表时间：
2017-06-01
期刊：
影响因子：
5.1
通讯作者：
Shao, Yuan
中科院分区：
文献类型：
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作者：
Jin, Xing-Wei;Liu, Bo-Ke;Shao, Yuan
本研究旨在探讨膀胱匀浆二次免疫C57 BL/6小鼠能否建立一种新的实验性自身免疫性膀胱炎（EAC）模型。将C57 BL/6小鼠用在完全弗氏佐剂（CFA）中的膀胱匀浆接种，并在2周后用在不完全弗氏佐剂（IFA）中的膀胱匀浆加强免疫用作EAC模型。用CFA或IFA中的磷酸盐缓冲盐水（PBS）免疫的小鼠用作对照。在初次免疫后4周测量排尿习惯和耻骨上骨盆疼痛阈值。然后检查膀胱与体重的比值以及炎性细胞因子和神经激肽1受体（NK 1 R）的表达。对膀胱进行组织学和免疫组织化学检查，还检测了肾、肝和肺的IL-1 β、IFN-γ和TNF-α产生。双重免疫小鼠对施加在骨盆区域上的压力广泛敏感（P < 0.001）。与单次免疫小鼠或对照组相比，双次免疫小鼠表现出更多的排尿频率、更低的每次排尿量、更高的膀胱/体重比以及炎性细胞因子（包括IL-1 β、IL-4、IL-6、IL-10、IFN-γ和TNF-α）表达的显著升高（均P < 0.05）。与其他三组相比，双重免疫组小鼠NK 1 R基因表达显著增加（P < 0.001）。非特异性免疫反应发生在肝脏，但比膀胱炎症弱得多。我们建立的C57 BL/6小鼠EAC模型能有效模拟BPS/IC的症状和病理生理特征，可广泛用于BPS/IC发病机制和治疗策略的研究。
The aim of this study is to identify whether vaccinating twice with bladder homogenate can establish a new model of experimental autoimmune cystitis (EAC) in C57BL/6 strain mice. C57BL/6 mice were vaccinated with bladder homogenate in complete Freund's adjuvant (CFA) and boost immunized with bladder homogenate in incomplete Freund's adjuvant (IFA) after 2 weeks were used as the EAC model. Mice immunized with phosphate-buffered saline (PBS) in CFA or IFA were used as the control. Micturition habits and suprapubic-pelvic pain threshold were measured 4 weeks after primary immunization. Bladder to body weight ratios and expression of inflammatory cytokines and neurokinin 1 receptor (NK1R) were then examined. Histologic and immunohistochemical examination of the bladder was carried out, and IL-1 beta, IFN-gamma, and TNF-alpha production by the kidneys, liver, and lungs was also tested. Double-immunized mice were extensively sensitive to pressure applied on the pelvic area (P < 0.001). Compared to single-immunized mice or controls, double-immunized mice showed more micturition frequency, lower urine output per micturition, higher bladder to body weight ratio, and significant elevation in the expression of inflammatory cytokines, including IL-1 beta, IL-4, IL-6, IL-10, IFN-gamma, and TNF-alpha (all P < 0.05). NK1R gene expression was significantly increased in double-immunized mice compared to the other three groups (P < 0.001). A nonspecific immune response occurred in the liver but was much weaker than bladder inflammation. Our dual immunization EAC model in C57BL/6 mice can effectively mimic the symptoms and pathophysiologic characteristics of BPS/IC and thus can be widely used to investigate the pathogenesis and therapeutic strategies of BPS/IC.