Effects of dental pulp stem cell transplantation on the long-term behavior and cAMP response element binding protein in neonatal rats with hypoxic ischemic brain damage

doi:10.3969/j.issn.2095-4344.2017.05.008

Abstract

Abstract:

BACKGROUND: cAMP response element binding protein (CREB) is a key protein of memory, which is closely related to long-term memory. It will provide a new way for the treatment of hypoxic ischemic brain damage (HIBD) to study the effects of dental pulp stem cells transplantation on the long-term behavior and CREB protein via the lateral ventricle in neonatal HIBD rats.
OBJECTIVE: To observe the changes in long-term behavior and CREB protein expression in neonatal HIBD rats after human dental pulp stem cell transplantation, thereby providing scientific evidence for clinical treatment of neonatal HIBD.
METHODS: Thirty-six healthy 7-day-old Sprague-Dawley rats were randomly divided into normal, HIBD and cell transplantation group. The hypoxic ischemic brain damage models were established in the brain damage and cell transplantation groups. Twenty-four hours after HIBD, human dental pulp stem cells were injected into the left lateral cerebral ventricle of rats in the cell transplantation group, totally 3×10⁶ living cells. Equal volume of normal saline was injected into the left lateral cerebral ventricle of rats in the normal control and HIBD groups.
RESULTS AND CONCLUSION: The average time to seek water, the average escape latency and escape distance of the human dental pulp stem cells group were significantly shorter than those of hypoxic ischemic brain injury group (P < 0.01), but longer than those in the normal group (P < 0.01). Nissl staining showed that the cells in the hippocampal CA1 region in human dental pulp stem cells group were more regular, the number of cells was significantly higher than that of hypoxic ischemic brain injury group, but still significantly less than that in the normal group (P < 0.05). Immunohistochemical staining results showed that the number of CREB positive cells in human dental pulp stem cells group was significantly higher than those in HIBD group, but still significantly less than those in the normal group (P < 0.01). It is suggested that human dental pulp stem cells transplantation could promote the expression of CREB protein in the hippocampal CA1 region, to improve the long-term learning and memory ability of hypoxic ischemic neonatal rats, and thus repair HIBD.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Dental Pulp, Cyclic AMP Response Element-Binding Protein, Tissue Engineering

CLC Number:

R394.2

Wang Ai, Mu Qing-jie, Wang Xiao-li, Yan Shao-zhen, Qu Peng-yu, Wang Hai-yu, Hu Wen-ting. Effects of dental pulp stem cell transplantation on the long-term behavior and cAMP response element binding protein in neonatal rats with hypoxic ischemic brain damage[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(5): 701-706.

Figures/Tables 2

2.2 实验动物数量分析 36只新生大鼠均进入结果分析，中途无脱落。 2.3 空间学习记忆能力 2.3.1 八臂迷宫觅水检测结果缺氧缺血性脑损伤组大鼠平均觅水时间显著长于正常组(Q=189.21，P < 0.01)。人牙髓干细胞组平均觅水时间较缺氧缺血性脑损伤组显著减(Q=129.17，P < 0.01)，仍高于正常组(Q=60.04，P < 0.01)。经过一段时间训练后，缺氧缺血性脑损伤组大鼠找到3个有水臂的路线要比正常组的路线要复杂，而人牙髓干细胞组大鼠的线路图较缺氧缺血性脑损伤组大鼠简化，但仍较正常组大鼠复杂(图2)。 2.3.2 Morris水迷宫检测结果缺氧缺血性脑损伤组大鼠平均逃避潜伏期显著长于正常组(Q=15.51，P < 0.01)。人牙髓干细胞组大鼠平均逃避潜伏期较缺氧缺血性脑损伤组显著缩短(Q=8.04，P < 0.01)，仍高于正常组(Q=7.48，P < 0.01)。经过一段时间训练后，缺氧缺血性脑损伤大鼠找到平台的路线要比正常组的复杂，而人牙髓干细胞组大鼠的线路图较缺氧缺血性脑损伤组大鼠简化，但仍较正常组大鼠复杂(图3)。 2.4 海马CA1区的细胞凋亡正常组大鼠脑组织海马区神经元细胞排列整齐，形态规则，尼氏体呈大颗粒，且数量较多；缺氧缺血性脑损伤组大鼠脑组织损伤侧海马区神经元细胞排列不整齐，形态不规则，神经元数量显著少于正常组(Q=99，P < 0.05)；人牙髓干细胞组大鼠脑组织损伤侧海马区神经元细胞排列较整齐，神经元数量显著多于缺氧缺血性脑损伤组(Q=30.5，P < 0.05)，但仍显著少于正常组(Q=68.5，P < 0.05；图4)。 2.5 海马CA1区CREB表达免疫组织化学染色结果显示，正常组见大量CREB+细胞表达，与正常组相比，缺氧缺血性脑损伤组与人牙髓干细胞组CREB+细胞数量减少(Q=79.17，P < 0.01；Q=34.75，P < 0.01)，人牙髓干细胞组CREB+细胞数显著多于缺氧缺血性脑损伤组(Q=44.42，P < 0.01；图5)。"

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