Nesfatin-1及其mRNA在大鼠生长轴中的表达

Expression of Nesfatin-1 and its mRNA in growth axis of rats

【目的】探讨Nesfatin-1在大鼠生长轴(下丘脑、腺垂体、肝脏、胰腺、肌肉)中的定位及其mRNA的表达情况。【方法】对雌性大鼠进行深度麻醉灌注后,采集下丘脑、腺垂体、肝脏、胰腺、腓肠肌肌肉组织,放入4%多聚甲醛溶液中固定,采用免疫组化PV-9000二步法、DAB显色技术检测Nesfatin-1在生长轴上的分布,另取雌性大鼠断头处死,取相同组织用荧光定量PCR方法检测其中Nesfatin-1mRNA的表达情况。【结果】Nesfatin-1主要分布于下丘脑的室旁核、背内侧核、室周核、外侧区、弓状核、视上核、腺垂体、胰腺的胰岛和腺泡及肝脏与肌肉中。平均光密度分析显示,Nesfatin-1在室周核和胰岛的表达量显著地高于其他组织(P<0.05),在胰腺腺泡、肝脏及肌肉中的表达量则显著地低于其他组织(P<0.05),在下丘脑背内侧核、外侧区和室旁核中的表达量显著高于视上核、弓状核和腺垂体(P<0.05)。Nesfatin-1mRNA在胰腺中表达量最高,腺垂体次之,下丘脑和肝脏较低,在胰腺和腺垂体的表达量显著高于下丘脑和肝脏,但下丘脑与肝脏差异不显著(P>0.05);在腓肠肌中未检测出Nesfatin-1mRNA的表达。【结论】Nesfatin-1及其mRNA在生长轴中的广泛表达,表明Nesfatin-1可能对大鼠生长发育具有调控作用。

[Objective] In the paper,expression of Nesfatin-1 mRNA in growth axis of rats including hypothalamus,pituitary, liver, pancreas, and muscle was explored. [Method] After deep anesthesia perfu- sion, hypothalamus, adenohypophysis, liver, pancreas, and gastrocnemius muscle of female rats were collect- ed and fixed in 4% paraformaldehyde solution. Then,immuno-histochemical PV-9000 2-step method and DAB staining technique were used to detect the distribution of Nesfatin-1. Other female rats were beheaded and same tissues were collected and the expression was measured using fluorescence quantitative polymer- ase chain reaction （PCR）. [Result] Nesfatin-1 mainly distributed in the hypothalamic paraventricular nu- cleus （ PVN）, dorsomedial nucleus （ DMH ）, periventricular nucleus （ PeN）, lateral hypothalamic area （LHA） ,arcuate nucleus（ARC）, supraoptic nucleus（SON）, pituitary, pancreas islet and acinar cells, liver, and muscle. Average optical density of Nesfatin-1 in PeN and pancreatic islet was significantly higher （P〈 0.05） than in other tissues while that in pancreatic acini, liver and muscle was significantly lower （P〈 0.05）. The expression in hypothalamic DMH, LHA and PVN was significantly different from in SON, ARC and anterior pituitary. The expression level of Nesfatin-1 mRNA in pancreas was the highest followed by pituitary,while the expression in liver and hypothalamus was the lowest. No expression of Nesfatin-1 mRNA was observed in muscle. [Conclusion] Nesfatin-1 and its mRNA widely distribute in the growth axis, indicating that Nesfatin-1 may participate in regulation of growth and development in rats.