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山东大学学报 (医学版) ›› 2021, Vol. 59 ›› Issue (5): 82-89.doi: 10.6040/j.issn.1671-7554.0.2021.0029

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干预Sonic Hedgehog信号通路对宫内发育迟缓新生大鼠学习记忆能力的影响

卢游,且迪,伍晋辉,杨凡   

  1. 四川大学华西第二医院儿童保健科, 四川 成都 610041
  • 发布日期:2021-06-01
  • 通讯作者: 杨凡. E-mail:yangfan_hx2th@163.com
  • 基金资助:
    国家重点研发计划子课题项目(2019YFC0840702)

Effects of the intervention of Sonic Hedgehog signaling pathway on learning and memory ability in rats with intrauterine growth restriction

LU You, QIE Di, WU Jinhui, YANG Fan   

  1. Department of Child Health Care, West China Second University Hospital, Sichuan University, Chengdu 610041, Sichuan, China
  • Published:2021-06-01

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摘要： 目的 探讨干预Sonic Hedgehog(Shh)信号通路对宫内发育迟缓(IUGR)新生大鼠学习记忆能力的影响及其可能机制。 方法 应用低蛋白饮食法建立IUGR大鼠模型,将其子鼠随机分成模型组、Shh信号通路抑制剂cyclopamine组(CYC组)和Shh信号通路激活剂purmorphamine组(PUR组),另选取同期正常幼鼠作为正常对照组,每组20只。分组干预后,于30日龄时行Morris水迷宫实验检测新生大鼠学习与记忆能力;免疫荧光BrdU/NeuN双标法检测大鼠海马组织增殖水平;TUNEL染色观察海马组织细胞凋亡情况;分光光度法检测海马组织Caspase-3活性;Western blotting检测大鼠海马区Shh信号通路相关蛋白Smoothened(Smo)、Gli-1和Bcl-2蛋白表达水平。 结果 与正常对照组比较,模型组大鼠学习记忆能力以及海马神经元增殖水平降低(P<0.05),海马组织细胞凋亡水平和Caspase-3活性增加(P=0.028, P=0.029),同时海马组织中Smo、Gli-1和Bcl-2等蛋白表达水平降低(P=0.017, P=0.026, P=0.031)。与模型组比较,PUR组大鼠学习记忆能力以及海马神经元增殖水平增加(P<0.05),海马组织细胞凋亡水平和Caspase-3活性降低(P=0.033, P=0.045),同时海马组织中Smo、Gli-1和Bcl-2等蛋白表达水平升高(P=0.041, P=0.016, P=0.033);而CYC组大鼠以上各项指标与模型组比较则进一步恶化,差异有统计学意义(P<0.05)。 结论 激活Shh信号通路可通过促进海马神经元增殖、抑制细胞凋亡,而改善IUGR新生大鼠学习记忆能力。

关键词: Shh信号通路, 宫内发育迟缓, 学习记忆能力, 神经元增殖, 细胞凋亡

Abstract: Objective To investigate the effects of the intervention of Sonic Hedgehog(Shh)signaling pathway on learning and memory ability in rats with intrauterine growth restriction(IUGR)and its possible mechanism. Methods The rat models of IUGR were established by feeding with low protein diet during pregnancy, and their newborn rats were divided into model group, Shh signaling pathway inhibitor cyclopamine group(CYC group)and Shh signaling pathway activator purmorphamine group(PUR group), 20 rats in each group. In addition, 20 normal newborn rats in the same period were selected as the normal control group. After intervention, Morris water maze test was performed to detect the learning and memory ability of newborn rats at the age of 30 days. Immunofluorescence BrdU/NeuN double labeling method was used to detect the proliferation level of hippocampal neurons. TUNEL staining was used to observe the level of cell apoptosis in hippocampus. Spectrophotometry was used to detect the activity of Caspase-3 in hippocampus. Western blotting was used to evaluate the expression levels of Shh signaling pathway related protein Smoothened(Smo), Gli-1 and Bcl-2 in hippocampus. Results Compared with the normal control group, the ability of learning and memory and the proliferation level of hippocampal neuron in the model group were decreased(P<0.05), the apoptosis level of hippocampus and the activity of Caspase-3 were increased(P=0.028, P=0.029), while the protein expression levels of Smo, Gli-1 and Bcl-2 were reduced in hippocampal tissue(P=0.017, P=0.026, P=0.031). Compared with the model group, the ability of learning and memory and the proliferation level of hippocampal neuron in the PUR group were increased(P<0.05), the apoptosis level of hippocampus and the activity of Caspase-3 were decreased(P=0.033, P=0.045), and the protein expression levels of Smo, Gli-1 and Bcl-2 in the hippocampus tissue were increased(P=0.041, P=0.016, P=0.033). However, compared with the model group, the above indicators of rats in the CYC group were worse, and the difference was statistically significant(P<0.05). Conclusion Activation of Shh signaling pathway can improve the learning and memory ability of IUGR newborn rats by promoting the proliferation of hippocampal neurons and inhibiting cell apoptosis.

Key words: Sonic Hedgehog signaling pathway, Intrauterine growth restriction, Learning and memory ability, Neuronal proliferation, Apoptosis

中图分类号: 

  • R722.1
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