小鼠慢性骨盆疼痛综合征模型与机械痛阈及自噬水平的相关性探索

发布时间：2018-03-28 20:29

  本文选题：慢性骨盆疼痛综合征　切入点：CBL/小鼠　出处：《中国实验动物学报》2017年03期

【摘要】：目的构建慢性骨盆疼痛综合征前列腺炎(chronic prostatitis/chronic pelvic pain syndrome,CP/CPPS)C57BL/6小鼠模型,探索其机械痛阈和自噬相关微管轻链蛋白LC3和底物蛋白p62表达水平随造模时间的变化规律,为CP/CPPS疼痛及自噬水平研究提供动物实验依据。方法将36只雄性C57BL/6小鼠随机均分为空白组、对照组和模型组,模型组小鼠皮下多点注射大鼠前列腺蛋白提取液和完全弗式佐剂的混悬液,建立CP/CPPS小鼠模型。通过HE染色观察前列腺病理变化,运用Von Frey纤维丝测定骨盆区域机械痛阈,通过免疫组化染色检测LC3和p62表达水平,运用Image Pro Plus 6.0软件计算平均光密度。结果 HE染色可见模型组小鼠出现慢性前列腺炎,表现为不同程度的上皮增生和淋巴细胞侵润,且实验后第6个月前列腺出现上皮内瘤(prostatic intraepithelial neoplasia,PIN),表现为基底膜消失和细胞核异形性明显等,空白组和对照组则表现为正常组织学形态。与空白组及对照组相比,模型组机械痛阈随造模时间延长逐渐降低[初始痛阈值为(0.35±0.154)g,第22周为(0.008±0.000)g],差异有显著性(P0.05)。其LC3和p62表达水平逐渐增高[LC3,p62平均光密度值分别为,第1个月:(2.767±0.464)%,(2.872±1.642)%;第6个月:(13.501±1.900)%,(9.070±0.490)%],差异有显著性(P0.05)。结论成功建立了CP/CPPS模型,且造模后第6个月出现PIN。模型组小鼠机械痛阈随造模时间的延长逐渐降低,LC3和p62表达逐渐增高,表明CP/CPPS炎症微环境促进疼痛产生及加剧,并提高小鼠前列腺自噬水平,与PIN的发生发展密切相关。
[Abstract]:Objective to establish a model of chronic prostatitis/chronic pelvic pain pain C57BL / 6 mice with chronic prostatitis/chronic pelvic pain syndrome, and to explore the changes of mechanical pain threshold and the expression level of autophagocyte-associated microtubule light chain protein (LC3) and substrate protein (p62) in mice with chronic pelvic pain syndrome (CPPS / CPPS / C57BL / 6). Methods 36 male C57BL/6 mice were randomly divided into blank group, control group and model group. CP/CPPS mouse model was established by subcutaneous injection of rat prostatic protein extract and suspension of complete Freund adjuvant. The pathological changes of prostate were observed by HE staining, and the mechanical pain threshold of pelvic region was measured by Von Frey filament. The expression levels of LC3 and p62 were detected by immunohistochemical staining, and the mean optical density was calculated by Image Pro Plus 6.0 software. Results chronic prostatitis was found in the model group by HE staining, with different degrees of epithelial hyperplasia and infiltration of lymphocytes. After 6 months, prostatic intraepithelial neoplasia was found in the prostatic prostate, showing the disappearance of the basement membrane and the apparent heteromorphism of the nucleus, while the normal histologic morphology was observed in the blank group and the control group, compared with the blank group and the control group. In the model group, the mechanical pain threshold decreased gradually with the prolongation of the modeling time [the initial pain threshold was 0.35 卤0.154 g, the initial pain threshold was 0.008 卤0.000g at the 22nd week], and the difference was significant (P 0.05). In the first month, 2.767 卤0.464% and 13.501 卤1.900 卤9.070 卤0.490%, respectively, in the first month (2.872 卤1.642). Conclusion the CP/CPPS model was successfully established, and the mechanical pain threshold of the model group gradually decreased with the prolongation of the model time. The expression of LC3 and p62 increased gradually in the model group. The results showed that the inflammatory microenvironment of CP/CPPS promoted the generation and exacerbation of pain and increased the level of prostatic autophagy in mice, which was closely related to the occurrence and development of PIN.
【作者单位】： 广西医科大学生命科学研究院;桂林医学院生物技术学院;
【基金】：国家自然科学基金(项目批准号:81472414)
【分类号】：R-332;R697.33
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本文编号：1677918