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目的评估沙美特罗调控间充质干细胞(MSC)促进小鼠脂多糖(LPS)诱导的肺损伤修复的作用及机制。方法体外试验以不同浓度沙美特罗(0.1、1和10μmol/L)刺激人骨髓来源MSC,收集作用不同时间(0、3、6、12和24 h)后的MSC,行聚合酶链反应,测定白介素-1受体拮抗剂(IL-1ra)的信使RNA(mRNA)表达。动物试验选取C57BL/6J小鼠32只,随机分为4组,每组8只。对照组:予磷酸盐缓冲液气管内注射;损伤组:LPS气管内注射;移植组:MSC移植和LPS气管内注射;沙美特罗刺激组:沙美特罗刺激后的MSC移植和LPS气管内注射。小鼠经口气管插管滴注5 mg/kg的LPS造模,造模前24 h尾静脉移植5×10~5个MSC,造模后24 h处死小鼠后行支气管肺泡灌洗,灌洗液离心,上清冻存待测蛋白浓度,下层细胞进行分类计数。另取小鼠肺组织行苏木精-伊红染色,光镜下观察。结果不同浓度的沙美特罗刺激MSC后,均能增加IL-1ra mRNA表达。1μmol/L沙美特罗刺激MSC 3、6、12和24 h,IL-1ra m RNA表达均有升高,刺激6 h后IL-1ra m RNA表达升高达到峰值,同0 h相比差异有统计学意义(P<0.01)。动物试验发现,沙美特罗刺激组肺泡灌洗液蛋白浓度、白细胞计数和中性粒细胞计数较移植组进一步降低(P<0.05);组织学损伤也较移植组和损伤组减轻。结论沙美特罗刺激MSC后静脉移植可减轻小鼠LPS诱导的肺损伤,这一作用与沙美特罗促进MSC旁分泌IL-1ra有关。
Objective To evaluate the effect and mechanism of salmeterol regulation of mesenchymal stem cells (MSCs) on the repair of lung injury induced by lipopolysaccharide (LPS) in mice. METHODS: MSCs derived from human bone marrow were stimulated with different concentrations of salmeterol (0.1, 1 and 10 μmol / L) in vitro. The MSCs collected at different time points (0, 3, 6, 12 and 24 h) Messenger RNA (mRNA) expression of interleukin-1 receptor antagonist (IL-1ra) was measured. Twenty-two C57BL / 6J mice were randomly divided into 4 groups (8 in each group). Control group: intraperitoneal injection of phosphate buffered saline; injury group: LPS intratracheal injection; transplantation group: MSC transplantation and LPS tracheal injection; salmeterol stimulation group: MSC transplantation after salmeterol stimulation and LPS tracheal injection . Mice were intratracheally instilled with 5 mg / kg LPS via oral intratracheal intratracheal injection, 5 × 10 ~ 5 MSCs were transplanted into caudal vein 24 h before model establishment, and mice were sacrificed 24 h after model establishment. Bronchoalveolar lavage Centrifuge wash, the supernatant stored protein concentration to be tested, the lower cells for classification and counting. Another mouse lung tissue hematoxylin-eosin staining, light microscopy. Results Different concentrations of salmeterol stimulated the expression of IL-1ra mRNA. 1 μmol / L salmeterol stimulated the expression of IL-1ra m RNA at 3, 6, 12 and 24 h after MSCs stimulation, and the expression of IL-1 m RNA increased to a peak at 6 h after stimulation Statistical significance (P <0.01). Animal experiments found that salmeterol stimulated group alveolar lavage fluid protein concentration, white blood cell count and neutrophil count further decreased compared with the transplantation group (P <0.05); histological damage than the transplantation group and injury group reduced. Conclusions Salmeterol stimulates the transplanted MSC to reduce the LPS-induced lung injury in mice. The effect of salmeterol on the secretion of IL-1ra by MSCs.