移植骨髓间充质干细胞减轻脂多糖诱导的小鼠急性肺损伤

目的 观察移植骨髓间充质干细胞 （MSCs） 对脂多糖 （LPS） 诱导小鼠急性肺损伤 （ALI）的治疗修复作用。方法 全骨髓培养法培养小鼠骨髓MSCs;细胞免疫化学染色鉴定MSCs特异表面标记;小鼠咽后壁吸入LPS制造小鼠肺损伤;尾静脉注射引入MSCs;称重计算肺水肿指数;肺组织切片HE染色观察组织病理改变;ELISA检测肺泡灌洗液和肺组织匀浆中IL-1β含量;Brdu（5-Bromo-2-Deoxyuridine）标记供体MSCs,免疫组织化学染色及双染色观察移植细胞的迁移和分化状态。结果 培养的MSCs细胞表面标记CD44阳性,而造血系表面标记CD34阴性。吸入LPS后,小鼠出现典型的肺损伤病理改变,肺水肿指数和肺组织匀浆IL-1β含量明显增加。标记的MSCs移植入同种异体的肺损伤小鼠,其肺部出现标记的MSCs,并表达上皮细胞标志抗原-细胞角蛋白（CK）。治疗后小鼠的肺水肿指数和肺组织匀浆IL-1β含量下降。结论 外源性MSCs移植到肺损伤小鼠体内,可迁移至肺损伤部位,并表达上皮细胞标志;减轻肺水肿程度,减少炎症因子释放。     

骨髓间充质干细胞移植治疗重症急性胰腺炎肺损伤

BACKGROUND:A large number of studies have confirmed that bone marrow mesenchymal stem cells can couple with the circulation of the blood to other organs, promote pancreatic tissue repair injury and reduce pulmonary fibrosis, which have certain therapeutic effects on pancreas and lung injuries. OBJECTIVE:To study the therapeutic effect on severe acute pancreatitis-associated lung injury in rats after the transplantation of bone marrow mesenchymal stem cells. METHODS:Animal models of severe acute pancreatitis-associated lung injury were prepared in rats via retrograde injection of 4% sodium taurocholate. Sprague-Dawley rats were randomized into three groups and received bone marrow mesencnymal stem cell injection via the tail vein in transplantation group, the same volume of normal saline in control group, or no treatment in normal groups. All the treatments in each group were performed 24 hours after modeling. Twenty-four hours after transplantation, hematoxylin-eosin staining of the pancreatic and lung tissues was performed. mRNA expressions of tumor necrosis factor-α and interleukin-1β in pancreatic and lung tissues were detected. ELISA kit was used to detect levels of serum C-reactive protein and tumor necrosis factor-α. RESULTS AND CONCLUSION:After modeling, under hematoxylin-eosin staining, there were a large number of inflammatory cells infiltrating in the damaged pancreatic tissues, accompanied by incomplete acinar structures, seriously destroyed lobular structures, alveolar fusion in the lung tissues, thickening of the alveolar walls, and a large amount of inflammatory cells infiltrating in the alveoli. These findings indicated successful modeling of severe acute pancreatitis-associated lung injury in rats. After cell transplantation, the number of infiltrated inflammatory cells in the damaged pancreatic tissue was reduced, with clear lobular structures and no bleeding from the acini; the structure of lung tissues was clear, with complete alveolar walls, and the width of alveolar space was reduced. Immunohistochemical results showed that transplanted DAPI-labeled bone marrow mesenchymal stem cells were aggregated in the pancreas and lung tissue, and uneven distributed in the damaged area. No DAPI expression in the pancreas and lung tissue was found in the control group, indicating transplanted bone marrow mesenchymal stem cells migrated into the damaged pancreas and lung tissue through the blood circulation, to further repair the damage area. RT-PCR test results showed that compared with the control group, bone marrow mesenchymal stem cell transplantation significantly reduced the levels of tumor necrosis factor-α and interleukin-1β in the pancreatic and lung tissues (P < 0.05). Higher levels of C-reactive protein and tumor necrosis factor-α were found in the control group compared with the normal group (P < 0.01), while the lower levels were obtained in the control group (P < 0.05). To conclude, our findings suggest that bone marrow mesenchymal stem cell transplantation is an effective therapy for severe acute pancreatitis-associated lung injury, and its mechanism may be associated with the reduction of inflammatory reactions and translation into the pancreas and lung tissue.     

骨髓间充质干细胞对新生鼠高氧急性肺损伤作用的研究

目的探讨骨髓间充质干细胞(MSCs)在新生鼠高氧急性肺损伤中的作用。方法体外复苏大鼠骨髓间充质干细胞,95%氧气7d天制备新生大鼠高氧急性肺损伤模型。动物模型制备后,将40只新生大鼠随机分为实验组(经尾静脉注入含1×105MSCs的PBS)20只,对照组(经尾静脉注入等量的PBS)20只。以正常新生大鼠作为空白组20只。注入细胞后24h各组处死10只,48h各组处死10只,取肺脏切片、检测湿干重比值(W/D)及髓过氧化物酶(MPO)。结果 24h、48h实验组病理切片损伤均较对照组减轻;24h实验组W/D及MPO均低于对照组,差异有显著意义(P0.05);48h实验组W/D及MPO均明显低于对照组,差异有极显著性意义(P0.01)。结论骨髓间充质干细胞对新生鼠高氧急性肺损伤可起保护作用。     

人脐带间充质干细胞移植治疗急性肺损伤

BACKGROUND:Studies have shown that human umbilical cord mesenchymal stem cells can improve pulmonary ventilation function by reducing inflammations. OBJECTIVE:To observe the therapeutic effect of human umbilical cord mesenchymal stem cell transplantation on acute lung injury. METHODS:Thirty Sprague-Dawley rats were randomized into normal group, model group and experimental group. Rats in the latter two groups were used to establish animal models of acute lung injury by intratracheal instillation of lipopolysaccharide. One hour after modeling, rats in the experimental group were intratracheally administered human umbilical cord mesenchymal stem cell suspension (0.1 mL, 1×10⁶ cells), and those in the other two groups were given normal saline in the same dose intratracheally. Twenty-four hours after treatment, the pathological changes of lung tissue were observed using hematoxylin-eosin staining; the wet and dry weight ratio of the lung tissue and the levels of serum interleukin-1 and interleukin-8 were detected. RESULTS AND CONCLUSION:Compared with the normal group, the wet and dry weight ratio of the lung tissue and the levels of serum interleukin-1 and interleukin-8 were significantly increased in the model group (P < 0.05), while compared with the model group, these levels were significantly decreased in the experimental group (P < 0.05). Hematoxylin-eosin staining results showed clear alveolar space structure with complete alveolar septum in the normal group. In the model group, the alveolar septum was markedly thickened, and there was visible pulmonary capillary hyperemia, edema, as well as a large amount of inflammatory cell infiltrations in the pulmonary capillaries and alveolar space. Edema fluid rich in proteins was observed in a part of the pulmonary alveoli, and an extensive transparent membrane formed in the alveolar space. In the experimental group, the alveolar structure was clear, but the alveolar septum became thickened, and red blood cells and a small amount of infiltrated inflammatory cells were leaked from the pulmonary interstitial tissue. In conclusion, human umbilical cord mesenchymal stem cell transplantation for treatment of acute lung injury can reduce inflammatory factor levels and alleviate lung injury.     

间充质干细胞治疗急性胰腺炎的研究进展

间充质干细胞具有强大的免疫调节能力和促进组织修复的巨大潜能。在急性胰腺炎的发病过程能够抑制T、B淋巴细胞及多种炎症因子,并能"归巢"至损伤胰腺,促进损伤胰腺组织修复,参与了急性胰腺炎的病理生理过程。本文将对间充质干细胞在急性胰腺炎病程中的作用作一综述。     

人脐带间充质干细胞移植治疗大鼠急性肺损伤

背景：间充质干细胞具有免疫调节特性,脐带间充质干细胞因其特有的优势,将在急性肺损伤和急性呼吸窘迫综合征的临床应用中有着光明的前景。 目的：探讨脐带间充质干细胞移植对内毒素性大鼠急性肺损伤模型的保护作用。 方法：将48只健康雄性SD大鼠随机均分为正常对照组、急性肺损伤组和脐带间充质干细胞移植组。后两组采用经气管内滴注内毒素建立急性肺损伤模型。成功造模1 h后,脐带间充质干细胞移植组,经气管内滴注脐带间充质干细胞混悬液,正常对照组和急性肺损伤组同法予以等量生理盐水。分别在干预后24,72 h,观察肺组织病理改变,检测病理组织评分、肺组织干湿质量比、髓过氧化物酶活性及血浆白细胞介素6、白细胞介素10及肿瘤坏死因子α水平。 结果与结论：脐带间充质干细胞移植可以减轻内毒素诱导的急性肺损伤模型的损伤程度;脐带间充质干细胞移植组在各时间点与急性肺损伤组比较,病理损伤评分、肺干湿质量比、肺组织髓过氧化物酶活性、血浆白细胞介素6和肿瘤坏死因子α水平均明显降低,血浆白细胞介素10水平明显升高。说明脐带间充质干细胞对内毒素性急性肺损伤模型有保护作用;其保护机制可能为脐带间充质干细胞移植维持肺内炎性递质和抗炎递质平衡。     

同种异体骨髓间充质干细胞移植治疗重症急性胰腺炎相关肺损伤

背景：目前骨髓间充质干细胞移植治疗各种炎症性疾病研究甚多,但在重症急性胰腺炎相关器官损伤干预方面研究甚少。 目的：观察同种异体骨髓间充质干细胞移植对重症急性胰腺炎相关肺损伤大鼠的干预作用。 方法：采用全骨髓差异贴壁法培养获得骨髓间充质干细胞。100只SD大鼠随机取32只为假手术组,仅翻动轻柔胰腺;另68只制作重症急性胰腺炎肺损伤动物模型,并分为干预组和对照组,每组再分为4个时间点。分别经尾静脉注入1×10⁹ L^-1浓度骨髓间充质干细胞悬液和同体积生理盐水。干预组每个时间点随机取1只注射CM-DiI标记的骨髓间充质干细胞悬液用于细胞示踪研究。 结果与结论：逆行胆胰管注射建模能早期诱发重症急性胰腺炎及相关肺损伤,炎症因子及E-选择素表达明显增高,并且胰腺和肺的损伤程度随时间延长而加重;移植荧光标记的干细胞后肺组织可见红色荧光出现,并随时间增长而增多;干预组肺组织损伤情况均较对照组各时间点减轻,血清淀粉酶及炎症递质肿瘤坏死因子α、白细胞介素1β较对照组各时间点下降。干预组肺组织中E-选择素表达较对照组下降。提示同种异体骨髓间充质干细胞移植能有效保护肺血管内皮细胞,减轻重症急性胰腺炎相关肺损伤。     

骨髓来源间充质干细胞培养上清液减轻脂多糖诱导的小鼠急性肺损伤

 目的:观察经尾静脉输注骨髓间充质干细胞培养上清液（MSCs CdM）对脂多糖(LPS)诱导的小鼠急性肺损伤的治疗作用及其机制。方法:采用全骨髓培养法分离纯化骨髓间充质干细胞,传至第3代时观察细胞形态,流式细胞术检测细胞表面标志,并且收集上清液用超滤离心管进行离心。30只BALB/c小鼠随机分为对照组、 LPS模型组和MSCs CdM治疗组。对照组腹腔内注射生理盐水（0.01 mL/g）,LPS组和MSCs CdM治疗组腹腔内注射LPS（5 mg/kg,0.01 mL/g）制备急性肺损伤模型。造模1 h后经尾静脉输注MSCs CdM（MSCs  CdM治疗组）或生理盐水 （LPS组或对照组）300 μL。6 h后处死小鼠,留取标本检测肺组织病理形态学、肺组织湿干重比（W/D）、支气管肺泡灌洗液（BALF）中蛋白含量、血清及BALF中细胞因子水平和肺组织中髓过氧化物酶(MPO)的活性。结果:与对照组比较,LPS处理后肺组织病理损伤严重,BALF中蛋白、血清肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)含量、肺组织中MPO活性及肺组织湿干重比均显著升高。与LPS组比较,MSCs CdM治疗组肺组织病理损伤程度减轻,BALF中蛋白、血清TNF-α和IL-6含量、肺组织中MPO活性及肺组织湿干重比均显著降低,而BALF中白细胞介素10（IL-10）和角质细胞生长因子（KGF）水平显著高于LPS组和对照组。结论:骨髓间充质干细胞培养上清液可有效减轻LPS诱导的急性肺损伤,其作用机制可能与其调节肺部TNF-α、IL-6、IL-10和KGF的水平有关。     

间充质干细胞治疗肺损伤的作用机制及研究进展

文题释义： 急性肺损伤：是各种直接和间接致伤因素导致的肺泡上皮细胞及毛细血管内皮细胞损伤,出现弥漫性肺间质及肺泡水肿,导致急性低氧性呼吸功能不全。以肺容积减少、肺顺应性降低、通气/血流比例失调为病理生理特征,其发展至严重阶段(氧合指数<200)被称为急性呼吸窘迫综合征。 外泌体：是细胞外泌囊泡中体积较小的一种,包含了复杂RNA和蛋白质的小膜泡(30-150 nm),所有培养的细胞类型均可分泌外泌体,且外泌体天然存在于体液中,间充质干细胞分泌的外泌体是近来研究的热点,其可以直接或者激活细胞内的信号通路进而发挥保护损伤细胞的作用。 背景：间充质干细胞疗法作为一种新兴的治疗方案,在治疗肺损伤方面具有很大的前景优势。 目的：综述间充质干细胞治疗肺损伤的作用和保护机制,以期为临床治疗肺损伤提供理论依据。 方法：检索万方、CNKI、PubMed、Web of Science等数据库2001年5月至2019年5月期间关于间充质干细胞应用于肺损伤的文献,检索词分别为“间充质干细胞,肺损伤,肺”和“mesenchymal stem cells,lung injury,pulmonary injury”。排除陈旧以及重复的观点,将检索到的文献进行整理,共纳入53篇文献来进行分析。 结果与结论：①梳理了间充质干细胞的定义、生物学特性及治疗肺损伤的机制,发现其可通过自身作用以及产生多种细胞因子和外泌体来治疗肺损伤;②总结了间充质干细胞治疗肺损伤的相关信号通路,例如PI3K/AKT信号通路、Wnt信号通路、NF-κB信号通路;③总结发现间充质干细胞与其他药物联合应用可以增强治疗肺损伤的效果,例如与促红细胞生成素、皮质类固醇联合应用等;④上述研究为临床上应用间充质干细胞治疗肺损伤提供了理论依据。 ORCID: 0000-0002-4555-9102(梁贵友) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

骨髓间充质干细胞对髋部骨折诱导的老年大鼠急性肺损伤的保护机制

目的　探讨骨髓间充质干细胞（bone marrow-derived mesenchymal stem cells,BMSCs）对老年单侧髋部骨折大鼠急性肺损伤（acute lung injury, ALI）的保护作用。 方法　将15月龄雌性WISTAR大鼠分为对照组（不做任何处理）、模型组（右髋部骨折+生理盐水）、治疗组（右髋部骨折+BMSCs）。治疗组于造模后经尾静脉注射1 ml BMSCs(2.5×106/ml),模型组给予等体积生理盐水。创伤后24 h进行肺组织病理评分,ELISA法检测检测肺组织MPO含量、肺及血液炎症因子（TNF-α、IL-6、IL-10）水平。 结果　与对照组相比,模型组肺部病理学评分、肺组织MPO水平、肺及血液炎症反应相关指标均明显升高（P<0.01）。BMSCs处理可明显降低肺组织病理学评分,肺组织MPO水平、肺及血液炎症反应相关指标下调,同时IL-10水平有更明显的增高（P<0.01）。 结论 髋部骨折可诱发老年大鼠急性肺损伤,BMSCs移植对老年髋部骨折大鼠肺损伤具有保护作用,其机制与缓解全身及肺部炎症反应有关。     

人脐带间充质干细胞防治新生大鼠急性肺损伤的作用

背景：研究发现间充质干细胞对成年动物急性肺损伤具有一定的防治作用。 目的：进一步验证人脐带间充质干细胞对内毒素所致的新生大鼠急性肺损伤的防治作用。 方法：选取7 d龄新生SD大鼠120只,随机分为3组。模型组和干细胞组大鼠腹腔注射内毒素3 mg/kg建立新生鼠急性肺损伤模型;生理盐水组大鼠腹腔注射0.1 mL生理盐水。30 min后干细胞组腹腔注射0.1 mL人脐带间充质干细胞(1×10⁶),生理盐水组和模型组大鼠腹腔注射等量生理盐水作对照。分别于6 h、1 d、2 d、4 d、7 d留取新生大鼠肺组织及血清标本,观察肺组织病理变化,ELISA方法测定血清中肿瘤坏死因子α及白细胞介素10的浓度及肺组织中髓过氧化物酶活性。 结果与结论：肺组织苏木精-伊红染色及髓过氧化物酶活性均提示模型组呈急性肺损伤改变,干细胞组造模后4 d及7 d时肺损伤比模型组轻。与模型组比较,干细胞组的抗炎因子白细胞介素10水平显著升高,而促炎因子肿瘤坏死因子α则明显减低,差异有显著性意义(P < 0.05)。说明用人脐带间充质干细胞早期细胞干预新生SD大鼠急性肺损伤,可以减轻肺部炎性病变,其主要机制可能为人脐带间充质干细胞通过免疫调节,平衡抗炎因子及促炎因子的表达,减轻肺损伤。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

骨髓间充质干细胞联合还原型谷胱甘肽对肺损伤小鼠的保护

背景：大量实验证明骨髓间充质干细胞治疗肺部疾病或改善肺损伤方面具有良好的效果,其治疗作用主要以减少炎性反应为主。 目的：观察骨髓间充质干细胞移植联合还原型谷胱甘肽对博来霉素诱导的小鼠肺损伤的保护效果。 方法：取1只雄性NOD/SCID小鼠制备骨髓间充质干细胞,并观察其形态、表型。将64只雌性NOD/SCID小鼠按随机数字表法分为对照组、模型组、骨髓间充质干细胞组和骨髓间充质干细胞+还原型谷胱甘肽组,每组16只。对照组气管内注入生理盐水,模型组气管内注入博来霉素,骨髓间充质干细胞组气管内注入博来霉素2 h后尾静脉内注入培养的骨髓间充质干细胞,骨髓间充质干细胞+还原型谷胱甘肽组气管内注入博来霉素2 h后尾静脉注入培养的骨髓间充质干细胞与还原型谷胱甘肽,7 d后处死动物,按照试剂盒说明测定肺组织匀浆肿瘤坏死因子α、白细胞介素1β、丙二醛水平,同时留取肺组织进行病理检查,确认骨髓间充质干细胞联合还原型谷胱甘肽对肺损伤的保护效果。 结果与结论：雄性小鼠骨髓间充质干细胞呈上皮细胞样,其CD34、CD45阴性表达,CD10、CD13、CD44阳性表达。雌性小鼠中,与对照组相比,模型组肿瘤坏死因子α、白细胞介素1β及丙二醛水平上升,骨髓间充质干细胞组和骨髓间充质干细胞+还原型谷胱甘肽组肿瘤坏死因子α、白细胞介素1β及丙二醛水平下降(P < 0.05),骨髓间充质干细胞+还原型谷胱甘肽组较骨髓间充质干细胞组下降更明显(P < 0.05)。病理切片显示,骨髓间充质干细胞+还原型谷胱甘肽组肺损伤较模型组及骨髓间充质干细胞组轻。以上结果表明骨髓间充质干细胞联合还原型谷胱甘肽能更有效保护博来霉素诱导的肺损伤。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程