骨髓间充质干细胞来源外泌体对盆腔炎模型大鼠氧化应激和炎症反应的影响

Effects of exosomes derived from bone marrow mesenchymal stem cells on oxidative stress and inflammatory response in rats with pelvic inflammatory disease

目的研究骨髓间充质干细胞(BMSCs)源性外泌体(Exosomes)对盆腔炎(PID)模型大鼠氧化应激和炎症反应的影响。方法通过全骨髓粘附法体外分离,培养和纯化正常SD大鼠的骨髓组织中的BMSCs。采用ExoQuick试剂法从BMSCs上清液中分离提纯Exosomes。按随机数字表法将60只雌性SD大鼠分为模型组、治疗组、左氧氟沙星组、对照组、空白组和假手术组6组,每组10只。对各组大鼠进行子宫组织HE染色,外周血白细胞测定,血清炎症因子检测,氧化应激状态评估。结果分离培养大鼠BMSCs,并成功提取了BMSCs源性Exosomes。治疗组大鼠子宫组织HE染色显示炎症反应较模型组和对照组明显减轻。治疗组大鼠外周血白细胞数、中心粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞均较模型组出现降低;且治疗组大鼠血清TNF-α、IL-1β、IL-6和IL-8的浓度水平值也均较模型组出现降低(P<0.05)。氧化应激指标检测显示治疗组大鼠SOD和T-AOC表达水平均较模型组出现升高,而MDA表达水平则降低(P<0.05)。治疗组大鼠除中心粒细胞数和血清IL-1β浓度水平值要高于左氧氟沙星组,其余外周血白细胞、血清炎症因子和氧化应激指标比较无统计学差异(P>0.05)。结论BMSCs源性Exosomes对盆腔炎模型大鼠子宫具有保护作用,其机制可能通过抑制氧化应激和炎症反应发挥作用。

Objective To study the effect of Exosomes derived from bone marrow mesenchymal stem cells on oxidative stress and inflammatory response in rats with pelvic inflammatory disease.Methods BMSCs from the bone marrow of normal SD rats were isolated,cultured and purified in vitro by total bone marrow adhesion.Exosomes were isolated from BMSCs supernatant by ExoQuick reagent method.According to the random number table method,60 female SD rats were divided into 6 groups(10 rats in each group).Model group(PID model rats without any special treatment).Treatment group(1 ml of exosomes diluted solution(100μg/mL)was injected intraperitoneally on the third day after the operation for 3 consecutive days).Levofloxacin group(PID model rats were given levofloxacin 60mg/kg/day by gavage on the 3rd day after surgery for 3 consecutive days).Control group(PID model rats were injected with corresponding dose of normal saline).Blank group(normal feeding of rats without any treatment)and Sham operation group(rats were only been opened and closed the abdomen).HE staining of uterine tissues,determination of peripheral blood leukocytes,detection of serum inflammatory factors and assessment of oxidative stress status were performed in each group.Results BMSCs of rats were isolated and cultured,and Exosomes derived from BMSCs were successfully extracted.HE staining showed that the inflammatory response of uterus tissue in the treatment group was significantly less than that in the model group and the control group.The number of white blood cells,central granulocytes,lymphocytes,monocytes and eosinophils in peripheral blood of rats in the treatment group were all decreased compared with those in the model group(P<0.05).The serum levels of TNF-α,IL-1β,IL-6 and IL-8 in the treatment group were also lower than those in the model group(P<0.05).The detection of oxidative stress index showed that the expression level of SOD and T-AOC in the treatment group was higher than that in the model group,while the expression level of MDA was lower(P<0.05).Except for the number of neutrophils and serum IL-1β concentration,there were no significant differences in peripheral blood leukocytes,serum inflammatory factors and oxidative stress indexes between the treatment group and Levofloxacin group(P>0.05).Conclusion Exosomes derived from BMSCs has a protective effect on the uterus of pelvic inflammatory model rats,and its mechanism may play a role by inhibiting oxidative stress and inflammatory response.