干细胞来源外泌体对早产仔鼠运动影响的研究

doi:10.11852/zgetbjzz2021-1253

摘要/Abstract

摘要： 目的研究人脐血间充质干细胞(Hu-MSCs)来源的外泌体对宫内感染仔鼠运动能力的影响及其机制,为治疗宫内感染及早产儿运动功能损害提供新思路。方法 90只孕鼠随机分为3组[实验组、内毒素(LPS组)和对照组,每组各30只]。孕15日,实验组孕鼠腹腔注射0.5 ml 1×10⁶ng/μl 外泌体悬液、LPS组与对照组腹腔注射等量生理盐水溶液;孕17日,实验组、LPS组宫内注射100 μl 内毒素(LPS,25 μg/μl)溶液,对照组宫内注射等量生理盐水。孕18日时3组孕鼠中分别随机选取15只孕鼠断颈处死, ELISA法检测孕鼠羊水、仔鼠血清中IL-4、IL-10、IL-12的改变;每组剩余的15只孕鼠等待自然分娩,观察妊娠结局;随机选取10只仔鼠待其15日龄、30日龄时使用悬吊实验、改良BBB运动评分(BBB Scale)评估仔鼠运动能力。结果 1)实验组孕鼠羊水、仔鼠血清中IL-4及IL-10较LPS组明显升高,两组之间差异有统计学意义(t=8.26、3.97、7.66、4.81,P<0.05),实验组孕鼠羊水、仔鼠血清中IL-12较LPS组明显降低,差异有统计学意义(t=3.14、6.68,P<0.05); 2)悬吊实验中,LPS组15日龄、30日龄仔鼠抓握时间较对照组明显缩短,差异有统计学意义(t=8.166、16.283,P<0.05);实验组较LPS组抓握时间显著延长,差异有统计学意义(t=4.867、6.994,P<0.05)。LPS组15日龄、30日龄仔鼠的改良BBB评分显著低于对照组,差异有统计学意义(t=7.394、16.283,P<0.05),实验组15日龄、30日龄仔鼠的改良BBB评分显著高于LPS组,差异有统计学意义(t=4.096、7.004,P<0.05)。结论脐血间充质干细胞来源的外泌体可能通过降低孕鼠及仔鼠体内炎性反应改善早产仔鼠运动能力的损害,为宫内感染及早产儿运动功能损害的提供新的治疗思路。

关键词: 外泌体, 间充质干细胞, 宫内感染, 运动能力, 炎性因子

Abstract: Objective To discuss the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (Hu-MSCs) on movement ability of the intrauterine infection offspring rats and its mechanism, so as to provide new ideas for treating intrauterine infection and motor function impariment. Methods Ninety pregnant rats were randomly divided into three groups [the experimental group, the lipopolysaccharide(LPS) group and the control group].On the 15th day of embryo(E15), dams of the experimental group were given 0.5 ml 1×10⁶ ng/μl intraperitoneal injection of exosomes, dams of the LPS group and the control group were given intraperitoneal injection of the same amount of physiological saline solution. On the 17th day of embryo(E17), dams of the experimental group and the LPS group were given intrauterine injection of 100 μl 25 μg/μl lipopolysaccharide (LPS) solution, and dams of the control group were given intrauterine injection of the same amount of normal saline. On the 18th day of embryo(E18), fifteen pregnant rats were randomly selected from the three groups and killed. Then ELISA method was used to check the change in the IL-4, IL-10 and IL-12 of the amniotic fluid of the pregnant rats and serum of the offspring. The remaining 15 pregnant rats in each group gave birth naturally and their pregnancy outcomes were observed. And 10 offspring were selected randomly to evaluate their movement ability by using Suspension Test and Basso-Beatie-Bresnahan Scale (BBB Scale) once 15 days old and once 30 days old. Results 1) The levels of IL-4 and IL-10 in the amniotic fluid of the dams and serum of the offspring in the experimental group were significantly elevated from those in the LPS group (t=8.26, 3.97, 7.66, 4.81, P<0.05). The levels of IL-12 in the amniotic fluid of the dams and serum of the offspring in the experimental group were significantly reduced from those of the LPS group (t=3.14, 6.68, P<0.05). 2) In suspension test, the grasp time of the 15-day-old and 30-day-old offspring in the LPS group was significantly shorter than those of the control group(t=8.166,16.283, P<0.05), which were also significantly shorter than those of the experimental group (t=4.867, 6.994, P<0.05).The BBB score of the 15-day-old and 30-day-old offspring in the LPS group were significantly lower than those of the control group (t＝7.394, 16.283, P<0.05),which were also significantly lower than those of the experimental group (t=4.096, 7.004, P<0.05). Conclusion The exosomes derived from human umbilical cord blood mesenchymal stem cells (Hu-MSCs) may improve the impairment of premature offspring pup's motor function by reducing the inflammatory response in pregnant and offspring rats.

Key words: exosomes, mesenchymal stem cells, intrauterine infection, movement ability, inflammatory factors

中图分类号:

R722.6

王娟, 宗璐. 干细胞来源外泌体对早产仔鼠运动影响的研究[J]. 中国儿童保健杂志, 2022, 30(5): 504-508.

WANG Juan, ZONG Lu. Effects of exosomes derived from stem cells on the movement of premature rats[J]. Chinese Journal of Child Health Care, 2022, 30(5): 504-508.

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