室旁核p-ERK神经元参与胰淀素抑食作用

李国俊1 刘昊东2; 3 曹晓娟1 杜晨光1; 2; 3*

doi:10.16098/j.issn.0529-1356.2025.06.003

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解剖学报 ›› 2025, Vol. 56 ›› Issue (6) : 651-657. DOI: 10.16098/j.issn.0529-1356.2025.06.003

神经生物学

室旁核p-ERK神经元参与胰淀素抑食作用

李国俊¹ 刘昊东^2,3 曹晓娟¹ 杜晨光^1,2,3*

作者信息 +

Paraventricular nucleus p-ERK neurons involving in the anorectic effect of amylin

LI Guo-jun¹ LIU Hao-dong^2,3 CAO Xiao-juan¹ DU Chen-guang^1,2,3*

Author information +

文章历史 +

摘要

目的探讨胰淀素（amylin）通过下丘脑室旁核（PVN）ERK通路对小鼠摄食行为的调控作用。方法将54只C57BL/6J小鼠随机分为多组开展实验。首先采用免疫荧光检测4只小鼠脑内ERK和降钙素受体（CTR）神经元的分布。通过化学损伤法建立PVN神经元损伤模型，将16只小鼠分为4组（每组4只）：对照组（Ctrl）、鹅膏酸损毁组（IBO）、amylin组和amylin+IBO组，统计PVN区域ERK阳性神经元数量及摄食量。随后，将16只小鼠分为4组（每组4只）：Ctrl组、amylin组、ERK拮抗剂U0126组、amylin+U0126组，通过Realtime PCR检测PVN即刻早期基因（c-fos）表达水平。免疫荧光检测Ctrl组和amylin组小鼠脑内p-ERK阳性神经元分布（每组4只）。最后，通过侧脑室注射，将18只小鼠随机分为3组（每组6只）：Ctrl组、amylin组和amylin+受体拮抗剂AC187组，然后免疫荧光检测PVN区域p-ERK、神经型一氧化氮合酶（nNOS）及CTR免疫阳性神经元共表达，同步记录处理后3 h内的摄食量。结果 ERK阳性神经元主要分布于视交叉上核（SCN）、弓状核（ARC）和蓝斑核（LC），而ERK和CTR共表达阳性神经元特异性富集于PVN（P＜0.01）。PVN神经元损伤导致ERK阳性神经元数量减少（P＜0.001），且阻断了amylin的抑食作用（P＜0.001）。Amylin激活PVN区域c-fos基因表达（P<0.05），而该效应可被U0126抑制（P＜0.05）。侧脑室注射amylin后，PVN中p-ERK、nNOS和CTR三标阳性神经元数量增加（P＜0.05），添加AC187则阻断amylin的激活效应（P＜0.01），并逆转amylin的抑食效应（1 h P=0.2218，2 h P=0.2218，3 h P=0.6974）。结论 PVN中p-ERK神经元通过amylin受体依赖的ERK信号通路调控摄食行为。

Abstract

Objective To explore the regulatory effect of amylin on the feeding behavior of mice through the ERK pathway of paraventricular nucleus of the hypothalamus (PVN). Methods A total of 54 C57BL/6J mice were randomly divided into multiple groups for experiments. First, immunofluorescence was used to detect the distribution of ERK and calcitonin receptor (CTR) neurons in the brains of 4 mice. A chemical lesion model of PVN neurons was established, and 16 mice were divided into four groups (4 mice per group): control (Ctrl) group, ibotenic acid (IBO)-lesioned group, amylin group, and amylin+IBO group. The number of ERK-positive neurons in the PVN region and food intake were then measured. Subsequently, 16 mice were divided into four groups (4 mice per group), Ctrl group, amylin group, ERK antagonist U0126 group, and amylin + U0126 group. The expression level of the immediate early gene c-fos in the PVN was detected by Real-time PCR. Immunofluorescence was used to detect the distribution of p-ERK positive neurons in the brains of 4 saline-treated mice and 4 amylin-treated mice. Finally, 18 mice were randomly divided into three groups (6 mice per group) through intracerebroventricular injection, Ctrl group, amylin group, and amylin + receptor antagonist AC187 group. Then, triple immunofluorescence was used to detect the co-expression of p-ERK, neuronal nitric oxide synthase (nNOS), and CTR-immunopositive neurons in the PVN region, and food intake within 3 hours after treatment was recorded simultaneously. Results ERK-positive neurons were mainly distributed in the suprachiasmatic nucleus (SCN), arcuate nucleus (ARC), and locus coeruleus (LC), while ERK and CTR co-expressing positive neurons were specifically enriched in the PVN (P=0.0012). Lesion of PVN neurons led to a decrease in the number of ERK-positive neurons (P<0.001) and blocked the anorectic effect of amylin (P<0.001). Amylin activated the expression of the c-fos gene in the PVN region (P<0.05), and this effect was inhibited by U0126 (P<0.05). After intracerebroventricular injection of amylin, the number of triple-positive neurons for p-ERK, nNOS, and CTR in the PVN increased (P<0.05). Addition of AC187 blocked the activation effect of amylin (P<0.01) and reversed the anorectic effect of amylin (1 h, P=0.2218; 2 h, P=0.2218; 3 h, P=0.6974). Conclusion P-ERK neurons in the PVN regulate feeding behavior through an amylin receptor-dependent ERK signaling pathway.

导出引用

李国俊刘昊东曹晓娟杜晨光. 室旁核p-ERK神经元参与胰淀素抑食作用[J]. 解剖学报. 2025, 56(6): 651-657 https://doi.org/10.16098/j.issn.0529-1356.2025.06.003

LI Guo-jun LIU Hao-dong CAO Xiao-juan DU Chen-guang. Paraventricular nucleus p-ERK neurons involving in the anorectic effect of amylin[J]. Acta Anatomica Sinica. 2025, 56(6): 651-657 https://doi.org/10.16098/j.issn.0529-1356.2025.06.003

中图分类号： R322.8 S852.1

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