摘要
目的:构建过敏性哮喘小鼠模型,研究人脐带干细胞来源的外泌体治疗过敏性哮喘模型小鼠的有效性,并探讨外泌体发挥作用的分子机制。方法:本研究中BALB/c小鼠随机分为人脐带间充质干细胞外泌体(hUCMSCs-exos)治疗组、激素治疗对照组、哮喘小鼠模型对照组和正常小鼠对照组。① 模型构建方法:在第1天和第14天给予15只小鼠腹腔内注射0.2 ml含有卵清蛋白(OVA)和AL(OH)3的混合液构建小鼠模型;② 治疗方法:在第21到27天,OVA组每天将致敏小鼠放入有机玻璃箱内给予5% OVA雾化30 min;正常对照组(CON)采用PBS雾化吸入30 min;激素治疗对照组采用布地奈德雾化吸入治疗30 min,每只鼠平均吸入0.02 mg;hUCMSCs-exos治疗组采用人脐带间充质干细胞培养上清制备的浓度为0.7 mg/ml的外泌体每天雾化治疗30 min。③ 标本采取和检测方法:造模和治疗结束后,采用眼周静脉丛采血法采静脉血;取肺组织和肺灌洗液。利用ELISA方法检测静脉血中的IgE及IL-4的浓度;肺组织切片做HE染色;肺灌洗液进行白细胞总数和嗜酸性粒细胞计数。结果:hUCMSCs-exos干预治疗组的小鼠与模型对照组的小鼠相比炎症明显改善,可明显降低IL-4 (p < 0.05)和IgE (p < 0.01)的水平,BALF中的白细胞总数及嗜酸性粒细胞的指标也明显降低(p < 0.01),小鼠的肺组织炎症也有明显的改善;布地奈德治疗组的干预效果与hUCMSCs-exos治疗组的效果相似,但是从小鼠肺泡灌洗液嗜酸性粒细胞数量的组间比较,可见hUCMSCs-exos治疗组的降低更加明显(p < 0.01)。结论:本研究结果证明,人脐带间充质干细胞来源的外泌体治疗过敏性哮喘模型小鼠是有效的。
Objective: To study the effectiveness of exosomes derived from human umbilical cord mesenchymal stem cells in the treatment of allergic asthma mouse models constructed with OVA, and explore the molecular mechanism of exosomes. Methods: In this study, BALB/c mice were randomly divided into human umbilical cord mesenchymal stem cell exosome (hUCMSCs-exos) treatment group, hor-mone therapy control group, asthma mouse model control group, normal mouse control group. ① Model construction Methods: On day 1 and day 14, 15 mice were injected intraperitoneally with 0.2 ml mixture containing OVA and AL(OH)3 to establish mouse models. ② Treatment methods: On days 21 to 27, the sensitized mice in the OVA group were put into a plexiglass box and given 5% OVA atomization for 30 min every day;normal control group (CON) was atomized by PBS for 30 min;the control group was treated with budesonide atomized inhalation for 30 min, with an average in-halation of 0.02 mg per rat. In the hUCMSCs-exos treatment group, exosomes with a concentration of 0.7 mg/ml prepared by human umbilical cord mesenchymal stem cell culture supernatant were atomized for 30 min every day. ③ The methods of specimen taking and testing were as follows: Af-ter modeling and treatment, venous blood was collected by periocular venous plexus;taking lung tissue and lung lavage fluid. IgE and IL-4 concentrations in venous blood were detected by ELISA. Lung tissue sections were stained with HE. Total white blood cells and eosinophils were counted in lung lavage fluid. Results: Compared with mice in the model control group, the inflammation of mice in hUCMSCs-exos intervention group was significantly improved, the levels of IL-4 (p < 0.05) and IgE (p < 0.01) were significantly decreased, and the total number of white blood cells and eosinophils in BALF was also significantly decreased (p < 0.01). The mice also showed significant improvement in lung tissue inflammation. The intervention effect of the budesonide treatment group was similar to that of the hUCMSCs-exos treatment group, but the intergroup comparison of the percentage of eo-sinophils in the alveolar lavage fluid of mice showed a more significant reduction in the hUCM-SCs-exos treatment group (p < 0.01). Conclusion: The results of this study prove that human umbil-ical cord mesenchymal stem cell derived exosomes is effective in the treatment of allergic asthma model mice.
出处
《临床医学进展》
2023年第8期13736-13744,共9页
Advances in Clinical Medicine
