PPARγ基因沉默的人骨髓基质细胞对骨髓抑制小鼠造血功能的影响  

Analysis on the Effects of PPARγGene-silenced Human Bone Marrow Stromal Cells on Hematopoietic Function in Mice with Bone Marrow Suppression

作　　者：王雪梅 黄纯兰 周铁军 李玉娇 魏梦宇 陈燕 陈晓敏 王万玥 Wang Xuemei;Huang Chunlan;Zhou Tiejun(Department of Hematology,the Affiliated Hospital of Southwest Medical University,Luzhou 646099,China;Stem Cell Laboratory,the Affiliated Hospital of Southwest Medical University,Luzhou 646099,China;Department of Pathology,the Affiliated Hospital of Southwest Medical University,Luzhou 646099,China)

机构地区：[1]西南医科大学附属医院血液内科,泸州646099 [2]西南医科大学附属医院干细胞实验室,泸州646099 [3]西南医科大学附属医院病理科,泸州646099

出　　处：《华中科技大学学报（医学版）》2024年第1期6-12,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：四川省科技计划项目(No.2020YJ0339);四川省泸州市科技计划项目(No.2019-SYF-36)。

摘　　要：目的 观察过氧化物酶体增殖激活受体γ(peroxisome proliferator activated receptor-gamma, PPARγ)基因沉默的人骨髓基质细胞HS-5对骨髓抑制小鼠造血功能的影响,并初步探讨其可能的作用机制。方法 用X射线进行全身照射构建骨髓抑制小鼠模型,造模后2 h,将小鼠随机分为3组,分别为实验组(尾静脉注射PPARγ RNAi干扰的HS-5细胞)、对照组(尾静脉注射未行PPARγ RNAi干扰的HS-5细胞)、空白组(尾静脉注射等量的生理盐水),每组5只。各组于放疗前、放疗后24 h、放疗后1周、放疗后2周进行外周血常规检测。对HS-5细胞在体外进行成骨、成脂诱导,分为实验组(PPARγ RNAi干扰的HS-5细胞)、对照组(未干扰PPARγ的HS-5细胞)、空白组(未行成骨/成脂诱导分化的HS-5细胞),观察成骨/成脂染色情况。采用CCK-8实验检测PPARγ基因沉默的HS-5细胞对小鼠骨髓造血干细胞(hemopoietic stem cell, HSC)增殖的影响,分为实验组(PPARγ RNAi干扰的HS-5细胞经成骨诱导分化3 d后,与小鼠HSC共培养)、阳性对照组(50μmol/L PPARγ抑制剂处理的HS-5细胞经成骨诱导分化3 d后,与小鼠HSC共培养)、阴性对照组(未干扰PPARγ的HS-5细胞经成骨诱导分化3 d后,与小鼠HSC共培养)、空白组(小鼠HSC单独培养,不与HS-5细胞共培养)。结果 放疗后,各组小鼠血常规指标均呈先降低后升高趋势,放疗后1周,三组小鼠血小板、白细胞水平差异显著,且实验组>对照组>空白组(均P<0.05);放疗后2周,三组小鼠脂肪空泡面积百分比差异显著,且实验组<对照组<空白组(均P<0.05),经Pearson相关分析显示,血常规各指标与血清PPARγ表达水平呈负相关(均P<0.05),与脂肪空泡面积百分比呈负相关(均P<0.05)。在体外成骨/成脂诱导分化后,实验组与对照组相比,橙红色的细胞比例明显降低,红色钙结节比例明显增高;成骨分化诱导3 d后,实验组、阳性对照组、阴性对照组人骨髓基质细胞均与小鼠Objective To investigate the effects of peroxisome proliferator activated receptor-gamma(PPARγ)gene silencing in human bone marrow stromal cells(HS-5)on hematopoietic function in bone marrow-suppressed mice,and to explore the potential mechanisms involved.Methods A bone marrow-suppressed mouse model was established by whole-body X-ray irradiation.Two hours after modeling,the mice were randomly divided into three groups:experimental group(intravenous injection of PPARγRNAi-interfered HS-5cells through the tail vein),control group(intravenous injection of PPARγRNAi-uninterfered HS-5cells through the tail vein),and blank group(intravenous injection of an equal amount of saline through the tail vein),with 5mice in each group.Peripheral blood routine tests were performed before,24hours after,1week after,and 2weeks after radiotherapy.In vitro osteogenic and adipogenic induction was performed in cells,and the cells were divided into experimental group(PPARγRNAi-interfered HS-5cells),control group(PPARγ-uninterfered HS-5cells),and blank group(HS-5cells without osteogenic/adipogenic induction).Osteogenic/adipogenic staining was observed.The effects of PPARγgene-silenced HS-5cells on mouse bone marrow hematopoietic stem cells(HSCs)were detected by CCK-8proliferation assay.The groups included experi mental group(PPARγRNAi-interfered HS-5cells were co-cultured with mouse HSCs after 3days of osteogenic induction differentiation),positive control group(HS-5cells treated with 50μmol/L PPARγinhibitor were co-cultured with mouse HSCs after 3days of osteogenic induction differentiation),negative control group(PPARγRNAi-uninterfered HS-5cells were co-cultured with mouse HSCs after 3days of osteogenic induction differentiation),and blank group(Mouse HSCs were cultured alone without co-culturing with HS-5cells).Results After radiotherapy,the hematological parameters of mice in each group showed a decreasing trend initially,and then increased.One week after radiotherapy,there were significant differences in platelet and whit

关 键 词：过氧化物酶体增殖激活受体Γ 人骨髓基质细胞 骨髓抑制小鼠 造血功能 成骨/成脂分化 

分 类 号：R552[医药卫生—血液循环系统疾病]