摘要
目的:探究脂多糖诱导的炎症反应对年轻小鼠骨髓及脾脏中造血干/祖细胞衰老表型的影响。方法:(1)构建细菌脂多糖(lipopolysaccharides,LPS)诱导的急性炎症小鼠模型,流式细胞术检测LPS刺激后年轻小鼠骨髓和脾脏中造血干/祖细胞的百分率,以及外周血和脾脏中各类成熟细胞百分率;通过增殖标记物Ki67检测小鼠骨髓和脾脏中造血干/祖细胞增殖的变化;检测炎症刺激后脾脏中造血干/祖细胞CD45蛋白的表达变化;(2)分析野生型年轻小鼠(2月龄)和年老小鼠(24月龄)骨髓和脾脏中造血干/祖细胞,外周血和脾脏中各类成熟细胞的百分率;(3)生物信息分析LPS刺激诱导造血干细胞转录组的变化。结果:(1)与对照组小鼠相比,体内LPS刺激导致小鼠骨髓中造血干/祖细胞百分率显著升高(P<0.05),外周血和脾脏中髓系细胞百分率显著升高(P<0.05);(2)与对照组小鼠相比,体内LPS刺激导致小鼠脾脏重量和细胞数目增多(P<0.05),脾脏中造血干/祖细胞百分率增多(P<0.05);(3)LPS刺激促进小鼠骨髓和脾脏中的造血干/祖细胞增殖(P<0.05);(4)LPS刺激导致小鼠脾脏中造血干/祖细胞CD45蛋白的表达降低(P<0.01);(5)与年轻小鼠相比,年老小鼠脾脏重量增加(P<0.05),脾脏中的造血干/祖细胞百分率增多(P<0.01);(6)与年轻小鼠相比,年老小鼠骨髓中的造血干细胞百分率显著升高(P<0.01),外周血和脾脏出现髓系分化偏向(P<0.01);(7)LPS刺激促进造血干细胞氧化应激和凋亡信号通路激活。结论:LPS刺激诱导造血干/祖细胞出现增殖,偏向髓系分化,髓外造血以及氧化应激、凋亡信号通路激活等造血干/祖细胞衰老表型。
AIM:This study aim to investigate the effects of lipopolysaccharide(LPS)-induced inflammation on the aging phenotype of hematopoietic stem/progenitor cells(HSPCs)in the bone marrow(BM)and spleen of mice.METHODS:(1)Young(2-month old)wild-type(WT)mice were treated with LPS to establish an actue inflammation model.The percentage of HSPCs in the BM and spleen of mice after LPS stimulation,as well as the ratio of mature cells in peripheral blood(PB)and spleen,were analyzed using flow cytometry.The proliferation of HSPCs in the BM and spleen was evaluated by examining the expression of the proliferation marker Ki67.In addition,changes in CD45 expression on HSPCs in the spleen of mice following LPS exposure were investigated by flow cytometry.(2)The percentage of HSPCs in BM and mature cells in PB and spleen of both young(2-month old)and old(24-month old)WT mice were analyzed by flow cytometry.(3)The transcriptome changes of hematopoietic stem cells(HSCs)after LPS stimulation was performed by an in silico analysis.RESULTS:(1)Mice exposed to LPS exhibited a significant increase in the percentage of HSPCs in BM and a marked elevation in the percentages of myeloid cells in PB and spleen compared to the mice in control group(P<0.05).(2)LPS exposure resulted in increased spleen weight and cell counts(P<0.05),along with a higher percentage of HSPCs in the spleen compared to controls(P<0.05).(3)LPS stimulation promoted the proliferation of HSPCs in the BM and spleen(P<0.05).(4)The expression of CD45 was reduced on HSPCs from spleen of mice after LPS stimulation(P<0.01).(5)In comparison to young mice,aged mice showed an increase in spleen weight and a higher percentage of HSPCs in the spleen(P<0.05).(6)Aged mice,in comparison to young mice,demonstrated a significantly higher percentage of HSPCs in the BM and myeloid skewing in the PB and spleen(P<0.01).(7)The silico analysis revealed upregualtion of reactive oxygen species(ROS)and apoptosis signaling in HSPCs following LPS stimulation.CONCLUSION:Young HSPCs stimulated by LPS exhibited an increase in cell number,a bias towards myeloid differentiation,enhanced extramedullary hematopoiesis,and elevated levels of ROS and apoptosis,all of which collectively manifested the aging phenotype of HSPCs.
作者
白可
邹密
詹蔷
黄颖欣
鞠振宇
陈陟阳
BAI Ke;ZOU Mi;ZHAN Qiang;HUANG Yingxin;JU Zhenyu;CHEN Zhiyang(Key Laboratory of Regenerative Medicine of Ministry of Education,Institute of Aging and Regenerative Medicine,College of Life Science and Technology,Jinan University,Guangzhou 510632,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2024年第1期38-46,共9页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.92049112)
广东省自然科学基金面上项目(No.2022A1515011204)。
关键词
炎症
造血干细胞
衰老
髓外造血
髓系分化
inflammation
hematopoietic stem cells
aging
extramedullary hematopoiesis
myeloid differen‐tiation
