脐血单个核细胞移植联合高压氧治疗对新生大鼠缺氧缺血性脑损伤的影响

Effects of umbilical cord mononuclear cells transplantation combined with hyperbaric oxygen therapy on hypoxic-ischemic brain damage in neonatal rats

摘要:

目的 观察脐血单个核细胞(UCBMC)联合高压氧(HBO)治疗对缺氧缺血性脑损伤(HIBD)新生大鼠远期行为学及组织学的影响。方法 7 日龄 Sprague-Dawley 新生大鼠随机分为正常对照组、缺氧缺血性脑损伤(HIBD)组、UCBMC 移植组及 UCBMC 联合HBO(UCBMC+HBO)组。采用Rice 法制成HIBD 模型。HBO 于造模 3 h 进行,造模后 24 h 行 UCBMC 移植。采用Western blot 法观察联合治疗对 HIBD 大鼠脑内IL-1β、TNF-α 蛋白的影响,采用T 迷宫、放射性迷宫试验及尼氏染色法,观察联合治疗对 HIBD 大鼠远期行为学及HIBD 大鼠脑海马 CA1 区组织学的影响。结果 移植后24 h,UCBMC+HBO 组大鼠脑内 IL-1β、TNF-α 蛋白的表达水平均显著低于HIBD 组与 UCBMC 组(P<0.05)。HIBD 组学习记忆及运动能力均减退,海马CA1 区锥体细胞数减少,UCBMC+HBO 组大鼠学习记忆及运动能力均较UCBMC 组及HIBD 组有改善,海马CA1 区锥体细胞数显著多于UCBMC 组及HIBD 组(P<0.05)。结论 UCBMC 联合HBO 治疗可减轻HIBD 新生大鼠IL-1β、TNF-α 蛋白的表达,促进脑损伤的修复并改善远期行为学。

Abstract:

Objective To study the effects of umbilical cord monoculcear cells (UCBMC) transplantation combined with hyperbaric oxygen (HBO) therapy on the long-term behaviors and histology in neonatal rats after hypoxic-ischemic brain damage (HIBD). Methods Seven-day-old Sprague-Dawley rats were randomly assigned to four groups: normal control (CON), HIBD, UCBMC and UCBMC+HBO. HIBD was induced according to the Rice-Vannucci method. The rats in the UCBMC+HBO group were treated with HBO 3 hours after HIBD, followed by UCBMC transplantation 24 hours after HIBD. IL-1β and TNF-α protein levels were examined by Western blot analysis in the 4 groups. T-maze test and radial arm maze test were used to detect the long-term learning memory capability. Nissl staining was used to examine the histological changes of the hippocampal CA1 region. Results Twenty-four hours after transplantation, IL-1β and TNF-α protein levels in the UCBMC+HBO group were significantly reduced compared with the HIBD (P<0.01) and UCBMC groups (P<0.05). The study and memory capabilities were impaired, and the number of the pyramidal cells in the hippocampal CA1 region was reduced in the HIBD group. The study and memory capabilities were greatly improved and the number of pyramidal cells increased significantly in the UCBMC+HBO group compared with the UCBMC and HIBD groups (P<0.05). Conclusions UCBMC transplantation combined with HBO therapy could reduce the expression of IL-1β and TNF-α protein, improve long-term behaviors and alleviate brain damages in the hypoxic ischemic neonatal rats.