血红素加氧酶-1修饰的骨髓间充质干细胞减轻脂多糖诱导的肺微血管内皮细胞氧化应激  

Bone marrow-derived mesenchymal stem cells modified with heme oxygenase-1 attenuate lipopolysaccha-ride-induced oxidative stress in pulmonary microvascular endothelial cells

作　　者：陈旭昕 李虎明 唐俭 孟激光 韩志海 CHEN Xuxin;LI Huming;TANG Jian;MENG Jiguang;HAN Zhihai(Department of Pulmonary and Critical Care Medicine,the Sixth Medical Center,PLA General Hospital,Beijing 100048,China;College of Pulmonary and Critical Care Medicine,PLA General Hospital,Beijing 100091,China;不详)

机构地区：[1]中国人民解放军总医院第六医学中心呼吸与危重症医学科,北京100048 [2]中国人民解放军总医院呼吸与危重症医学部,北京100091

出　　处：《实用医学杂志》2021年第18期2339-2343,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:81300050);北京市自然科学基金项目(编号:7182163);海军总医院创新培育基金项目(编号:CXPY201417);解放军总医院第六医学中心创新培育基金(编号:CXPY201902)。

摘　　要：目的探讨血红素加氧酶-1(HO-1)修饰的骨髓间充质干细胞(MSC-HO-1)对脂多糖(LPS)刺激下肺微血管内皮细胞(PVEC)氧化应激的影响。方法实验分组:A组(PVEC+LPS),PVEC仅予LPS刺激;B组(PVEC/MSC+LPS),PVEC细胞与MSC共培养于Transwell体系过夜后予LPS刺激;C组(PVEC/MSC-HO-1+LPS),同上PVEC与MSC-HO-1共培养后予LPS刺激;D组(正常PVEC)。采用流式细胞仪及活性氧检测试剂盒,测PVEC内活性氧(ROS)含量;采用比色法及相应商品化检测试剂盒,测PVEC内脂质过氧化物(LPO)、丙二醛(MDA)水平及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)活性;用Western blot检测各组PVEC内HO-1表达变化。结果与D组相比,A组受LPS刺激后,出现明显氧化应激(P<0.05);与MSC或MSC-HO-1共培养均可明显减轻氧化应激(P<0.05),但MSC-HO-1共培养减轻氧化应激程度更为显著(P<0.05)。Western blot结果显示与MSC-HO-1共培养可明显上调PVEC内HO-1蛋白表达。结论MSC-HO-1显著改善LPS诱导的PVEC氧化应激失衡,可能机制与上调PVEC内HO-1表达有关。Objective To explore the influence of bone marrow-derived mesenchymal stem cells modified with heme oxygenase-1(MSC-HO-1)on oxidative stress in pulmonary microvascular endothelial cells(PVECs)following lipopolysaccharide(LPS)stimulation.Methods PVECs were divided into group A(PVEC+LPS),group B(PVEC/MSC+LPS),group C(PVEC/MSC-HO-1+LPS)and group D(PVEC).In groups B and C,PVECs were co-cultured with MSC or MSC-HO-1 overnight.LPS(0.1 mg/mL),as a stimulus,was thenadded into transwell co-culture system for 6 hours.Intracellular accumulation of ROS was measured using a commercial kit and flow cytometry.Activities of lipid peroxide(LPO),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH-PX)in PVECs were determined by using colorimetric assay.HO-1 protein expression of PVEC was analyzed by Western blot.Results As compared with group D,LPS-stimulation caused the imbalance of oxidative stress in PVECs.Co-culture with MSC or MSC-HO-1 significantly inhibited oxidative stress induced by LPS in PVECs,and the extent of inhibition in group C was more evident.Western blot analysis showed that coculture with MSC-HO-1 could significantly upregulate protein expression of HO-1 in PVECs.Conclusions MSCHO-1 attenuates lipopolysaccharide-induced oxidative stress in PVECs.The detailed mechanism might be associated with upregulation of HO-1 expression in PVECs.

关 键 词：间充质干细胞 脂多糖 血红素加氧酶-1 氧化应激 肺微血管内皮细胞 

分 类 号：R563.9[医药卫生—呼吸系统]