Nesfatin-1对内脏高敏感大鼠脑肠轴CRF-R1表达的影响

The effect of Nesfatin-1 on the expression of CRF-R1 in visceral hypersensitivity rats

目的探讨Nesfatin-1对内脏高敏感大鼠脑肠轴不同部位促肾上腺皮质释放激素受体1(CRF-R1)表达的影响。方法将新生2 d龄雄性Spragne-Dawley大鼠随机分为健康对照组和模型组,模型组大鼠通过母婴分离和醋酸灌肠相结合的方法制备。模型验证成功后,分为干预对照组、低浓度干预组、中浓度干预组和高浓度干预组,健康对照组不给予任何处理;干预对照组大鼠给予侧脑室注射生理盐水0.5μL;低浓度组、中浓度组和高浓度组大鼠分别给予侧脑室注射0.5、5和50μmol/L的Nesfatin-1 0.5μL,应用免疫组化(IHC)和ELISA方法检测内脏高敏感模型大鼠脑肠轴不同部位CRF-R1的表达。结果 ELISA检测结果显示,健康对照组、干预对照组大鼠分别与低浓度组、中浓度组、高浓度组大鼠结肠CRF-R1水平进行比较,差异均有统计学意义(P<0.01)。IHC检测结果显示,健康对照组、干预对照组分别与低浓度组、中浓度组、高浓度组在结肠组织、脊髓组织、脑组织中CRF-R1阳性表达之间的差异有统计学意义(P<0.05或0.01);Nesfatin-1不同浓度组大鼠脑肠轴CRF-R1阳性表达明显高于健康对照组、干预对照组,且脑组织中CRF-R1阳性表达最显著。结论 Nesfatin-1通过影响中枢和外周CRF-R1信号通路,参与内脏高敏感的形成及调控。

Objective To explore the effect of Nesfatin-1 on corticotropin releasing factor receptor-1（CRF-R1）expression in brain gut axis of visceral hypersensitivity rats. Methods Two days male Spragne-Dawley rats were divided randomly into healthy control group and model group.The model group was induced to visceral hypersensitivity by maternal separation and acetic acid enema methods. After the model validation succeed,the model group were divided randomly into intervention control group,low concentration intervention group,medium concentration intervention group and high concentration intervention group. The healthy control group was given nothing,the intervention control group was given 0.5 μL Saline,the low concentration group,medium concentration group and high concentration group were give 0.5,5 and 50 μmol/L Nesfatin-1respectively.We tested the expression of CRF-R1 in different parts of the brain gut axis by immunohistochemistry（IHC）and enzyme-linked immunosorbent assay（ELISA）methods. Results The enzyme-linked immunosorbent assay test results indicated the CRF-R1 levels in different groups significant differences（P〈0.01）. The immunohistochemistry test results also indicated the CRF-R1 expression levels in different groups were significantly different（P〈0.01）. Nesfatin-1 could increase the CRF-R1 expression levels at different concentration and the expression of CRF-R1 was obviously increase in brain.Conclusion Nesfatin-1 involved in the formation and the regulation of visceral hypersensitivity in IBS though influencing the CRF-R1 signalling between the central and peripheral nervous system.