骨髓间充质干细胞通过抑制髓系抑制细胞预防肺纤维化形成  

Bone marrow mesenchymal stem cells prevent pulmonary fibrosis through inhibiting myeloid-derived suppressor cells

作　　者：尹燕锋[1] 杨柳[1] 于璐[2] 王翼寅 陈睿[1] 周开华[3] 苏晓三[1] YIN Yanfeng;YANG Liu;YU Lu;WANG Yiyin;CHEN Rui;ZHOU Kaihua;SU Xiaosan(Biomedical Research Center,The Affiliated Calmette Hospital of Kunming Medical University,Kunming,Yunnan 650011,P.R.China;Department of Pathology,The Affiliated Calmette Hospital of Kunming Medical University,Kunming,Yunan 650011,P.R.China;Department of Respiratory Medicine,The Affiliated Calmette Hospital of Kunming Medical University,Kunming 650011,P.R.China)

机构地区：[1]昆明医科大学附属甘美医院生物医学实验中心,云南昆明650011 [2]昆明医科大学附属甘美医院病理科,云南昆明650011 [3]昆明医科大学附属甘美医院呼吸科,云南昆明650011

出　　处：《中国呼吸与危重监护杂志》2021年第5期339-344,共6页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：国家自然科学基金(31360223);云南省应用基础研究(昆医联合专项)(2014FZ055)。

摘　　要：目的探讨Gr-1^(+)CD11b^(+)髓系抑制细胞(MDSC)在博来霉素(BLM)诱导小鼠肺纤维化形成中的作用以及骨髓间充质干细胞(MSC)对肺纤维化治疗作用的可能机制。方法体外分离培养6周龄雄性BALB/c小鼠间充质干细胞(MSC)。将104只雌性BALB/c小鼠按随机数字表法分为3组:对照组(32只)气管内注入生理盐水(NS)后经尾静脉注射NS;BLM组(32只)气管内注入BLM后经尾静脉注射NS;MSC组(40只)气管内注入BLM后经尾静脉注射MSC(细胞总数1.5×106个)。于造模后第1、3、5、8、11、14、18、21、25、32天采集各组小鼠尾静脉血,流式细胞术检测外周血单个核细胞(PBMC)中Gr-1^(+)CD11b^(+)细胞含量。于造模后第3、8、18和32天各组均处死8只小鼠并摘取小鼠肺脏流式细胞术检测小鼠肺组织中Gr-1^(+)CD11b^(+)细胞含量,同时留取肺组织行Masson染色。提取MSC组小鼠肺组织的DNA,通过聚合酶链反应(PCR)-琼脂糖电泳法检测小鼠的性别决定基因(sry基因)。结果 MSC组第3和第8天时肺泡炎较BLM组减轻(分别为1.32±0.25和2.53±0.56;1.06±0.42和2.27±0.82,均P<0.01);MSC组第18和第32天时肺纤维化较BLM组减轻(分别为1.02±0.44和1.81±0.74;1.51±0.73和2.72±0.54,均P<0.05)。BLM组小鼠PBMC和肺组织中Gr-1^(+)CD11b^(+)细胞较对照组显著升高;MSC组小鼠PBMC和肺组织中Gr-1^(+)CD11b^(+)细胞较BLM组明显降低(P<0.05)。PCR检测结果显示,在MSC治疗后第12、24、48、72和96 h小鼠肺组织可以检测到sry基因。结论 MDSC参与BLM诱导的肺纤维化形成,MSC可以通过抑制MDSC预防肺纤维化形成。Objective To investigate the role of myeloid-derived suppressor cell(MDSC) in bleomycin(BLM)-induced pulmonary fibrosis and the possible mechanism of bone marrow mesenchymal stem cell(MSC) in therapy of BLM-induced pulmonary fibrosis. Methods Bone marrow mesenchymal stem cells(MSC) were harvested from 6-week old male BALB/c mice. One hundred and four female BALB/c mice were randomly divided into 3 groups. Mice in control(n=32) and BLM group were instilled with normal saline(NS) or BLM via trachea and NS were injected via tail vein on the1 st, 2 nd and 3 rd day after NS administration. Mice in MSC group(n=40) were instilled with BLM via trachea and MSC(total cell number=1.5×106) were injected via tail vein. On the 1 st, 3 rd, 5 th, 8 th, 11 th, 14 th, 18 th, 21 st, 25 th and 32 nd day after BLM administration, the percentage of Gr-1^(+)CD11 b^(+) cells in peripheral blood mononuclear cell(PBMC) was detected by flow cytometry. Eight mice from each group were killed on the 3 rd, 8 th, 18 th and 32 nd day after BLM administration, the percentage of Gr-1^(+)CD11 b^(+) cells in the lung tissue was detected by flow cytometry. Meanwhile,the lung tissue specimens were stained with Masson. The sry gene of Y chromosome was detected by polymerase chain reaction(PCR). Results Compared with BLM group, MSC transplantation significantly reduced pulmonary inflammation in MSC group [(1.32±0.25) vs.(2.53±0.56);and(1.06±0.42) vs.(2.27±0.82), respectively, (P<0.01)]. Likewise,MSC transplantation significantly reduced pulmonary fibrosis and deposition of collagen as compared with BLM group[(1.02±0.44) vs.(1.81±0.74), and(1.51±0.73) vs.(2.72±0.54), respectively, (P<0.05)]. The percentage of Gr-1^(+)CD11 b^(+) cells in the BLM group was significantly increased as compared with control group. Compared with BLM group, MSC transplantation significantly reduced Gr-1^(+)CD11 b^(+) cells in MSC group(P<0.05). The sry gene(201 bp) was detected in the lungs of female mice within 96 hours after MSC administration. Conclusions MDSC parti

关 键 词：肺纤维化 间充质干细胞 髓系抑制细胞 博来霉素 

分 类 号：R563.9[医药卫生—呼吸系统]