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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 investigate the effect of Nesfatin-1 on the expression of adrenocorticotropic hormone-releasing hormone receptor 1 (CRF-R1) in different parts of visceral hypersensitivity rat brain. Methods Male Sprague-Dawley rats of 2 days old were randomly divided into healthy control group and model group. The model rats were prepared by the combination of mother-infant separation and acetic enema. After successful model validation, the rats were divided into intervention control group, low concentration intervention group, medium concentration intervention group and high concentration intervention group, and the healthy control group received no treatment. The rats in the control group were injected intracerebroventricular injection with normal saline (0.5 μL) 0.5, 5 and 50 μmol / L Nesfatin-1 0.5 μL were intracerebroventricularly administered to rats in the middle, high and middle concentration groups, respectively. Immunohistochemistry (IHC) and ELISA were used to detect the visceral hypersensitivity Different parts of the expression of CRF-R1. Results The results of ELISA showed that the levels of CRF-R1 in colon of healthy control group and intervention control group were significantly lower than those of low concentration group, middle concentration group and high concentration group (P <0.01). The results of IHC showed that the difference of CRF-R1 positive expression between the control group, the control group and the low concentration group, the middle concentration group and the high concentration group in the colon, spinal cord and brain tissue were statistically significant (P <0.05 or 0.01). The expression of CRF-R1 in brain and intestine of Nesfatin-1 group was significantly higher than that in healthy control group and intervention group, and the expression of CRF-R1 in brain tissue was the most significant. Conclusion Nesfatin-1 participates in the formation and regulation of visceral hypersensitivity by affecting the CRF-R1 signaling pathway in the central and peripheral regions.