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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
[1] Sacchi C, Marino C, Nosarti C, et al. Association of intrauterine growth restriction and small for gestational age status with childhood cognitive outcomes: a systematic review and meta-analysis[J]. JAMA Pediatr, 2020, 174(8): 772-781.
[2] 刘爽, 冯琪, Guellec I, 等. 宫内及宫外生长对极早产儿远期神经系统发育的影响[J].中华新生儿科杂志, 2017, 32(3): 240. doi: 10.3760/cma.j.issn.2096-2932.2017.03.023. LIU Shuang, FENG Qi, Guellec I, et al. The effect of intrauterine and extrauterine growth on the long-term nervous system development of very premature infants[J]. Chinese Journal of Neonatology, 2017, 32(3): 240. doi: 10.3760/cma.j.issn.2096-2932.2017.03.023.
[3] 刘美娟, 龙鼎新. Hedgehog信号通路在胚胎发育过程中的调控作用[J]. 生命的化学, 2017, 37(2): 142-146. LIU Meijuan, LONG Dingxin. The regulation of Hedgehog signaling pathway in embryonic development[J]. Chemistry of Life, 2017, 37(2): 142-146.
[4] Yin S, Bai X, Xin D, et al. Neuroprotective effects of the Sonic Hedgehog signaling pathway in ischemic injury through promotion of synaptic and neuronal health[J]. Neural Plast, 2020, 2020: 8815195. doi: 10.1155/2020/ 8815195.
[5] 崔学文, 陆浩, 吕德民, 等. SHH修饰聚多巴胺涂层纤维蛋白支架对大鼠神经干细胞的影响[J].神经解剖学杂志, 2020, 36(1): 15-22. CUI Xuewen, LU Hao, LYU Demin, et al. The effect of SHH modified polydopamine-coated fibrin scaffold on rat neural stem cells[J]. Journal of Neuroanatomy, 2020, 36(1): 15-22.
[6] 杨艳, 俞生林. 宫内生长发育迟缓早产儿住院情况研究[J].中国儿童保健杂志, 2019, 27(6): 637-641. YANG Yan, YU Shenglin. Study on hospitalization of premature infants with intrauterine growth retardation[J]. Chinese Journal of Child Health Care, 2019, 27(6): 637-641.
[7] 应一博, 陈敏, 朱娟娟, 等. 内源性成体神经干细胞神经发生机制的研究进展[J].生物医学工程学杂志, 2018, 35(6): 986-992. YING Yibo, CHEN Min, ZHU Juanjuan, et al. Research progress in neurogenesis of endogenous adult neural stem cells[J]. Journal of Biomedical Engineering, 2018, 35(6): 986-992.
[8] 崔兆辉, 滕元君, 姜金, 等. Sonic Hedgehog信号通路在成年大鼠脊髓损伤后的表达[J].中国修复重建外科杂志, 2015, 29(5): 576-581. CUI Zhaohui, TENG Yuanjun, JIANG Jin, et al. The expression of Sonic Hedgehog signaling pathway after spinal cord injury in adult rats[J]. Chinese Journal of Reconstructive Surgery, 2015, 29(5): 576-581.
[9] Tichy J, Zinke J, Bunz B, et al. Expression profile of Sonic Hedgehog pathway members in the developing human fetal brain[J]. Biomed Res Int, 2015: 494269. doi: 10.1155/2015/494269.
[10] 高琳琳, 王军, 李子英, 等. 不同方法制备大鼠宫内发育迟缓模型的比较[J].中国比较医学杂志, 2018, 28(1): 1-7. GAO Linlin, WANG Jun, LI Ziying, et al. Comparison of rat intrauterine growth retardation models prepared by different methods[J]. Chinese Journal of Comparative Medicine, 2018, 28(1): 1-7.
[11] 于涛, 范玉颖, 王华. 大麻素受体1及神经黏附分子L1在宫内发育迟缓大鼠脑组织的表达[J]. 中华实用儿科临床杂志, 2013, 28(4): 301-303. YU Tao, FAN Yuying, WANG Hua. Expression of cannabinoid receptor 1 and neuroadhesion molecule L1 in brain tissue of rats with intrauterine growth retardation[J]. Chinese Journal of Applied Clinical Pediatrics, 2013, 28(4): 301-303.
[12] 罗小泉, 骆利平, 陈海芳, 等. Morris水迷宫检测大鼠记忆力方法的探讨[J]. 时珍国医国药, 2010, 21(10): 2667-2669. LUO Xiaoquan, LUO Liping, CHEN Haifang, et al. Discussion on Morris water maze for detecting memory in rats[J]. Lishizhen Medicine and Materia Medica Research, 2010, 21(10): 2667-2669.
[13] 刘文晶, 袁兆红. 宫内生长受限影响认知发育的机制研究进展[J]. 中国儿童保健杂志, 2019, 27(3): 281-284. LIU Wenjing, YUAN Zhaohong. Research progress on the mechanism of intrauterine growth restriction affecting cognitive development[J]. Chinese Journal of Child Health Care, 2019, 27(3): 281-284.
[14] 张勇, 王朝晖, 李燕晖. 宫内发育迟缓对早产儿体格发育的影响[J].中国儿童保健杂志, 2019, 27(12): 1345-1348. ZHANG Yong, WANG Chaohui, LI Yanhui. The effect of intrauterine growth retardation on physical development of premature infants[J]. Chinese Journal of Child Health Care, 2019, 27(12): 1345-1348.
[15] Arthurs OJ, Rega A, Guimiot F, et al. Diffusion-weighted magnetic resonance imaging of the fetal brain in intrauterine growth restriction[J]. Ultrasound Obstet Gynecol, 2017, 50(1): 79-87.
[16] 周厚妊, 刘治军, 解丽梅, 等. 宫内生长受限胎儿的主动脉峡部血流指数和围产儿不良结局的关系[J].中国超声医学杂志, 2019, 35(9): 824-826. ZHOU Houren, LIU Zhijun, XIE Limei, et al. The relationship between the aortic isthmus blood flow index of fetuses with intrauterine growth restriction and adverse perinatal outcomes[J]. Chinese Journal of Ultrasound in Medicine, 2019, 35(9): 824-826.
[17] 石怡芳. 超声诊断胎儿宫内生长迟缓的临床价值[J].山西医科大学学报, 2010, 41(12): 1082-1083. SHI Yifang. The clinical value of ultrasound diagnosis of intrauterine growth retardation in fetus[J]. Journal of Shanxi Medical University, 2010, 41(12): 1082-1083.
[18] Ruff CA, Faulkner SD, Rumajogee P, et al. The extent of intrauterine growth restriction determines the severity of cerebral injury and neurobehavioural deficits in rodents[J]. PLoS One, 2017, 12(9): e0184653. doi: 10.1371/journal. pone.0184653.
[19] 王粉, 丁晓春, 冯星. 高蛋白喂养对宫内发育迟缓大鼠脑髓鞘化过程及认知能力的影响[J]. 中华实用儿科临床杂志, 2013, 28(17): 1351-1354. WANG Fen, DING Xiaochun, FENG Xing. Effects of high-protein feeding on brain myelination process and cognitive ability in rats with intrauterine growth retardation[J]. Chinese Journal of Applied Clinical Pediatrics, 2013, 28(17): 1351-1354.
[20] Li PJ, Guo YQ, Ding PY, et al. Neuroprotective effects of a Smoothened receptor agonist against postoperative cognitive dysfunction by promoting autophagy in the dentate gyrus of aged rats[J]. Neurol Res, 2019, 41(10): 867-874.
[21] Hu Q, Li T, Wang L, et al. Neuroprotective effects of a smoothened receptor agonist against early brain injury after experimental subarachnoid hemorrhage in rats[J]. Front Cell Neurosci, 2017, 10: 306. doi: 10.3389/fncel.2016.00306.
[22] Mitchell N, Petralia RS, Currier DG, et al. Sonic hedgehog regulates presynaptic terminal size, ultrastructure and function in hippocampal neurons[J]. J Cell Sci, 2012, 125(Pt 18): 4207-4213.
[23] 张广慧, 冯金洲, 郭振委, 等. TRPC1沉默对脑缺血大鼠神经干细胞迁移的影响[J].中风与神经疾病杂志, 2017, 34(9): 782-785. ZHANG Guanghui, FENG Jinzhou, GUO Zhenwei, et al. The effect of TRPC1 silencing on the migration of neural stem cells in rats with cerebral ischemia[J]. Journal of Apoplexy and Nervous Diseases, 2017, 34(9): 782-785.
[24] 任登鹏, 李晓红, 涂悦, 等. 活性因子对大鼠创伤性脑损伤后内源性神经干细胞增殖和分化的影响[J].中华创伤杂志, 2016, 32(9): 843-847. REN Dengpeng, LI Xiaohong, TU Yue, et al. Effects of active factors on the proliferation and differentiation of endogenous neural stem cells after traumatic brain injury in rats[J]. Chinese Journal of Trauma, 2016, 32(9): 843-847.
[25] 王舒娅, 李晶, 杜元灏. Sonic Hedgehog信号通路及其对神经系统的影响[J].脑与神经疾病杂志, 2016, 24(8): 521-525. WANG Shuya, LI Jing, DU Yuanhao. Sonic Hedgehog signal pathway and its effect on the nervous system[J]. Journal of Brain and Nervous Diseases, 2016, 24(8): 521-525.
[26] Qin YT, Jiang M, Tuerxung N, et al. Sonic Hedgehog signaling pathway in Myelodysplastic Syndrome: abnormal activation and jervine intervention[J]. Gene, 2020, 754: 144881. doi: 10.1016/j.gene.2020.144881.
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