人脐带间充质干细胞条件培养基对宫内感染所致新生儿肺损伤的影响  

Effects of human umbilical cord mesenchymal stem cells conditioned medium on neonatal lung injury caused by intrauterine infection

作　　者：贺娟 王玲 黄蓉 周伟 吕回 陈晓文 陶莉 He Juan;Wang Ling;Huang Rong;Zhou Wei;Lyn Hui;Chen Xiaowen;Tao Li(Department of Neonatalogy,Guangzhou Women and Children’s Medical Center,Guangzhou 510623;Department of Neonatalogy,Xiangtan Municipal Central Hospital,Xiantan 411100,Hunan,China)

机构地区：[1]广州市妇女儿童医疗中心新生儿科,510623 [2]湘潭市中心医院新生儿科,湖南湘潭411100

出　　处：《中华生物医学工程杂志》2020年第2期102-109,共8页Chinese Journal of Biomedical Engineering

基　　金：广东省科技厅一般项目(2014A020212427);广州市医药卫生科技一般引导项目(20171A010271)。

摘　　要：目的研究人脐带间充质干细胞条件培养基(CM)对宫内感染致新生大鼠肺损伤模型的影响。方法选取孕龄20 d SD大鼠18只,随机分为3组(n=6)。NS组孕20、21 d连续2 d腹腔注入生理盐水1 ml,所生幼鼠随机选取30只新生鼠,于第3天腹腔注射生理盐水0.1 ml。LPS组孕20、21 d连续2 d予腹腔注入LPS(大肠杆菌内毒素,2.5 mg·kg^-1·d^-1),随机选取30只新生鼠,于第3天腹腔注入0.1 ml生理盐水。LPS+CM组孕20、21 d连续2 d予腹腔注入LPS(2.5 mg·kg^-1·d^-1),所生幼鼠随机选取30只新生鼠,于第3天腹腔注入CM 0.1 ml。3组新生鼠分别于7、14、21 d随机选取10只麻醉处死,观察各组3个时间点新生鼠体质量、肺重、肺重/体质量;HE染色观察新生鼠肺形态学和鼠肺辐射状肺泡计数(RAC);液相芯片技术检测各组新生鼠血清中IL-6、IL-10、TNF-α变化。结果LPS组新生鼠7、14、21 d体质量较NS组、LPS+CM组均减轻,3组新生鼠体质量随生后日龄不断增加(均P<0.05);LPS组新生鼠14、21 d肺重较NS组和LPS+CM组减轻,3组肺重随生后日龄不断增加(均P<0.05);LPS组新生鼠7 d肺重/体质量较NS组、LPS+CM组升高(均P<0.05),14 d仅较NS组升高(P<0.05),21 d仅较LPS+CM组降低(P<0.05),3组新生鼠肺重/体质量随生后日龄不断下降(均P<0.05)。肺组织HE染色显示NS组新生鼠肺组织结构完整,肺泡扩张均匀,肺泡间隔未见明显增厚,间隔内极少量炎细胞浸润,肺泡腔清晰;LPS组新生鼠肺泡数目减少,肺泡体积增大,肺泡间隔减少,部分肺泡间隔增厚,间隔内炎细胞浸润;LPS+CM组新生鼠肺泡数目、体积、肺泡间隔及间隔炎细胞浸润状态均较LPS组好转,损伤程度减轻。7 d LPS组新生鼠肺组织的RAC仅较NS组减少(P<0.05),14、21 d较NS组、LPS+CM组均减少(均P<0.05),3组新生鼠肺组织RAC随生后日龄不断增加(均P<0.05)。与NS组比较,LPS组新生鼠血清IL-6在7、14、21 d升高(均P<0.05),LPS+CM组则在7、21 d升高(均P<0.05),LPS+CObjective To investigate the effect of human umbilical cord mesenchymal stem cell(hu-MSC)conditioned medium(CM)on lung injury induced by intrauterine infection in rat models.Methods Eighteen SD rats with at 20 days of gestation were randomly divided into three groups(n=6 each).In the NS group,1 ml normal saline was injected intraperitoneally for 2 consecutive days on days 20 and 21 of pregnancy;30 of the neonatal off springs were randomly selected and injected with 0.1 ml normal saline intraperitoneally on day 3 postnatum.In the LPS group,lipopolysaccharide(an E.coli endotoxin)at a dose of 2.5 mg·kg^-1·d^-1 was intraperitoneally injected for 2 consecutive days on days 20 and 21 of pregnancy;30 of the neonatal off springs were randomly selected and injected with 0.1 ml normal saline intraperitoneally on day 3 postnatum.In the LPS+CM group,LPS at a dose of 2.5 mg·kg^-1·d^-1 was intraperitoneally injected for 2 consecutive days on days 20 and 21 of pregnancy;30 of the neonatal off springs were randomly selected and injected with 0.1 ml CM intraperitoneally on day 3 postnatum.The three groups of neonatal rats were randomly sacrificed in tens by anesthesia on days 7,14,and 21.The body mass,lung weight,and lung weight/body weight of the neonatal rats of each group were examined at the three time points.HE staining was used to study the morphology and radial alveolar count(RAC)of the lung in the rats.Luminex xMAP liquid phase microarray was used to detect the changes in serum IL-6,IL-10 and TNF-αin each group of neonatal rats.Results The body weight of neonatal rats in LPS group compared with the NS and LPS+CM groups on days 7,14 and 21;the body weight of neonatal rats in three groups increased with days after birth(all P<0.05).The lung weight of neonatal rats in LPS group was lower compared with NS and LPS+CM groups on days 14 and 21;the lung weight of neonatal rats in three groups increased with days after birth(all P<0.05).The lung weight/body weight in the neonatal rats in LPS group were higher than those in NS

关 键 词：间充质基质细胞 脐血干细胞 培养基 条件性 宫内感染 肺损伤 婴儿 新生 

分 类 号：R722.1[医药卫生—儿科] R-332[医药卫生—临床医学]