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     

骨髓间充质干细胞治疗早期骨性关节炎软骨T2值及相关因子的表达

背景：血清肿瘤坏死因子α和白细胞介素1β作为炎症递质,其水平与骨性关节炎病情相关。而骨髓间充质干细胞治疗骨性关节炎报道较少。 目的：用骨髓间充质干细胞治疗兔早期骨性关节炎,观察关节软骨T2值及血清肿瘤坏死因子α和白细胞介素1β水平并进行分析。 方法：将36只新西兰大白兔随机均分成对照组、模型组和治疗组。模型组兔仅建立骨性关节炎模型,治疗组在造模后第4周左膝关节腔内注射骨髓间充质干细胞;对照组膝关节腔内注射等量生理盐水。分别在治疗后第2周,1,2,3个月行左后膝关节核磁共振及血清肿瘤坏死因子α和白细胞介素1β水平检查。 结果与结论：与模型组比较,治疗组T2值降低,无关节腔积液;血清肿瘤坏死因子α和白细胞介素1β水平降低;软骨T2值变化与二者呈正相关。MRI能够反映早期关节软骨的生物学改变,可用于评价骨性关节炎治疗前后的改变;骨髓间充质干细胞在关节腔内可向软骨分化,为早期骨性关节炎患者提供了一种新的有效疗法。     

骨髓间充质干细胞尾静脉移植脊髓损伤大鼠脑源性神经营养因子及神经生长因子的表达

背景：骨髓间充质干细胞移植对脊髓损伤有治疗作用,但其机制尚不完全清楚。 目的：应用免疫组织化学方法观察骨髓间充质干细胞静脉移植损伤脊髓局部脑源性神经营养因子及神经生长因子的表达,分析骨髓间充质干细胞移植治疗大鼠脊髓损伤的作用途径。 方法：运用改良Allen法制备T10脊髓外伤性截瘫大鼠模型,假手术组6只,脊髓损伤组24只随机分为对照组和骨髓间充质干细胞移植组。骨髓间充质干细胞移植组、假手术组接受骨髓间充质干细胞单细胞悬液1 mL(1×10⁶ cells)自大鼠尾静脉缓慢注射移植,对照组静脉注射PBS 1 mL。 结果与结论：脊髓损伤后损伤局部的脑源性神经营养因子、神经生长因子表达增加,骨髓间充质干细胞静脉注射移植后能促进脊髓损伤局部脑源性神经营养因子、神经生长因子更进一步的表达,这可能是促进神经结构及神经功能恢复的因素之一。     

骨髓间充质干细胞移植修复慢性阻塞性肺病气道损伤

背景：间充质干细胞能够分化为肺实质细胞并参与肺部损伤的修复,为间充质干细胞在慢性阻塞性肺病中的应用提供了新的方法。 目的：观察骨髓间充质干细胞移植对大鼠慢性阻塞性肺病气道损伤的修复作用。 方法：将24只雌性大鼠随机分为4组：①骨髓间充质干细胞移植组(12只)：采用熏烟+脂多糖法建立慢性阻塞性肺病大鼠模型,于造模后第1天经尾静脉输注1 mL CM-Dil标记的骨髓间充质干细胞。②骨髓间充质干细胞对照组(4只)：在第1天和第14天经气管注入生理盐水300 μL,经尾静脉输注1 mL CM-Dil标记的骨髓间充质干细胞。③慢性阻塞性肺病模型组(4只)：采用熏烟+脂多糖法建立慢性阻塞性肺病大鼠模型,于造模后第1天经尾静脉输注1 mL PBS。④健康对照组(4只)：在第1天和第14天经气管注入生理盐水300 μL,经尾静脉输注1 mL PBS。在各组大鼠注射骨髓间充质干细胞后的第1,7,15,30天,进行病理学和血清学检测。 结果与结论：①苏木精-伊红染色结果显示：骨髓间充质干细胞移植组大鼠肺气肿和气道病变较慢性阻塞性肺病模型组轻,但较骨髓间充质干细胞对照组和健康对照组严重。②移植后第1天,骨髓间充质干细胞移植组外周血白细胞总数和中性粒细胞比例高于骨髓间充质干细胞对照组和健康对照组(P < 0.05),随着时间延长,骨髓间充质干细胞移植组白细胞总数和中性粒细胞比例不断降低。③移植后第1天,骨髓间充质干细胞移植组外周血白细胞介素10水平低于骨髓间充质干细胞对照组和健康对照组(P < 0.05),肿瘤坏死因子α和粒细胞集落刺激因子水平高于骨髓间充质干细胞对照组和健康对照组(P < 0.05)。随着移植时间的延长,骨髓间充质干细胞移植组外周血肿瘤坏死因子α水平不断降低,白细胞介素10水平不断升高,粒细胞集落刺激因子水平先升高再降低,其中第7天水平最高。④CM-Dil染色联合免疫组化检测提示,部分 CM-Dil阳性细胞同时CC16表达阳性。结果显示经尾静脉注射骨髓间充质干细胞能够改善慢性阻塞性肺病大鼠的肺部病理损伤,通过分化为气管黏膜上皮细胞以及参与免疫调节对气道进行修复。 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     

转化生长因子β1在大鼠骨髓间充质干细胞移植修复梗死心肌中的作用

背景：大量研究显示,骨髓间充质干细胞是通过旁分泌作用,促使移植部位大量肌纤维母细胞聚集,分泌大量胶原蛋白,从而使梗死后心脏得到有利修复,并改善心脏收缩舒张功能。 目的：探讨转化生长因子β1在大鼠骨髓间充质干细胞修复梗死心肌过程中的作用。 方法：①体外实验：模拟心肌梗死后微环境培养大鼠骨髓间充质干细胞,检测其分泌肿瘤坏死因子α、血小板衍生生长因子、转化生长因子β1浓度。②体内实验：将大鼠Brdu标记的骨髓间充质干细胞、转化生长因子β1、PBS分别移植到梗死后心肌内,免疫细胞化学染色、PCR、Wester-blot等方法检测移植后大鼠心肌胶原蛋白Ⅰ、Ⅲ形成的差异。 结果与结论：心肌梗死后微环境下培养大鼠骨髓间充质干细胞14 d可检测到培养基内有高浓度转化生长因子β1。移植骨髓间充质干细胞、转化生长因子β1大鼠心肌部位可检测到肌纤维母细胞的聚集及胶原蛋白Ⅰ、Ⅲ表达。结果显示大鼠骨髓间充质干细胞移植梗死心肌后,可促进胶原蛋白Ⅰ、Ⅲ的生成,从而改善心脏收缩功能,其中骨髓间充质干细胞所分泌的转化生长因子β1可能发挥重要作用。     

肿瘤坏死因子α刺激骨髓间充质干细胞表达及分泌肝细胞生长因子

背景：有研究表明,肿瘤坏死因子α可以刺激骨髓间充质干细胞发挥免疫抑制功能,并促进骨髓间充质干细胞表达肝细胞生长因子。 目的：观察肿瘤坏死因子α刺激大鼠骨髓间充质干细胞是否可以促进肝细胞生长因子的表达及分泌。 方法：全骨髓贴壁法分离、纯化SD大鼠骨髓间充质干细胞,传代至第三四代使用。以100 µg/L肿瘤坏死因子α预刺激骨髓间充质干细胞 5 h后弃去培养基,换上新鲜培养基作为实验组,以正常培养的骨髓间充质干细胞为空白组。采用倒置相差显微镜观察细胞形态;流式细胞仪鉴定骨髓间充质干细胞表面标记物CD29、CD34、CD44、CD45的表达;RT-PCR 、Western blot检测细胞肝细胞生长因子mRNA及蛋白表达;ELISA测定细胞上清液中肝细胞生长因子水平。 结果与结论：获得的骨髓间充质干细胞形态均一,呈典型的旋涡样生长。骨髓间充质干细胞表达CD29⁺99.45%,CD34^-97.91%、CD44⁺99.52%、CD45^-98.42%;骨髓间充质干细胞中肝细胞生长因子 mRNA及蛋白与上清液中肝细胞生长因子表达量呈时间依赖性增加,实验组肝细胞生长因子 mRNA及蛋白与上清液中肝细胞生长因子的表达量明显高于空白组(P < 0.01)。说明肿瘤坏死因子α刺激骨髓间充质干细胞可以有效促进肝细胞生长因子的表达及分泌。     

姜黄素对骨髓间充质干细胞诱导肾移植免疫耐受的影响

背景：已有研究显示在大鼠肾移植时连续给予1×10⁷细胞浓度的骨髓间充质干细胞可诱导免疫耐受,但临床上尚未发现姜黄素对肾移植具有免疫调节作用。目的：在对肾移植后的大鼠给予骨髓间充质干细胞的基础上,同时给予不同剂量姜黄素,观察其免疫耐受的效果。方法：建立大鼠肾移植模型,随机分为4组：纯肾移植组,无治疗措施;骨髓间充质干细胞组：关腹前左骼静脉注射1×10⁷/kg骨髓间充质干细胞,第2天起尾静脉注射等量细胞,共10 d;骨髓间充质干细胞+低/高剂量姜黄素组：除给予骨髓间充质干细胞外,分别以2,10 mg/kg姜黄素溶液灌胃,共10 d。免疫组织化学染色法检测各组肾组织中转化生长因子β1的蛋白表达水平,酶联免疫吸附法检测血清中白细胞介素2,6水平。结果与结论：大鼠经肾移植后,纯肾移植组中肾小管上皮细胞以及间质细胞的转化生长因子β1蛋白表达、白细胞介素2,6水平均高于其他各组(P < 0.05)。与骨髓间充质干细胞组比较,骨髓间充质干细胞+低/高剂量姜黄素组的转化生长因子β1的蛋白表达降低(P < 0.05),血清白细胞介素2,6水平均低于骨髓间充质干细胞组(P < 0.05)。结果证实,大鼠肾移植时同时给予姜黄素与骨髓间充质干细胞注射,可有效地进行免疫调节,抑制性免疫排斥反应的发生,保护肾功能。 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     

骨髓间充质干细胞移植治疗大鼠溃疡性结肠炎

背景：近年已有干细胞移植修复慢性肠炎受损组织的报道,但其治疗机制尚不明确。 目的：观察移植骨髓间充质干细胞在溃疡性结肠炎大鼠结肠的分布及分化情况,及对大鼠结肠黏膜的修复作用。 方法：取经鉴定的第3代大鼠骨髓间充质干细胞1×10⁶个,应用Hoechst 33342进行荧光标记,并将其经尾静脉移植入溃疡性结肠炎大鼠体内,分别于移植后3,7,14 d取材。 结果与结论：骨髓间充质干细胞移植后 3 d,大鼠结肠即可见荧光标记的细胞,移植后14 d,大鼠结肠荧光标记细胞仍较多,此时可见部分标记细胞表达细胞角蛋白。同时骨髓间充质干细胞移植后14 d,溃疡性结肠炎大鼠结肠黏膜白细胞介素10表达增高,肿瘤坏死因子α表达降低, 结肠黏膜的病理损伤明显好转。说明移植的骨髓间充质干细胞可迁移至结肠溃疡部位并分化为上皮细胞,同时可通过调节炎性因子的表达促进溃疡性结肠炎大鼠结肠黏膜损伤的修复。     

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。     

肿瘤坏死因子α预处理脐血间充质干细胞移植治疗心肌梗死

文章快速阅读：文题释义： 心肌缺血：是指心脏的血液灌注减少,导致心脏的供氧减少,心肌能量代谢不正常,不能支持心脏正常工作的一种病理状态。血压降低、主动脉供血减少、冠状动脉阻塞,可直接导致心脏供血减少;心瓣膜病、血黏度变化、心肌本身病变也会使心脏供血减少,心肌缺血对心脏和全身可能带来许多危害。目前针对这种疾病治疗主要的作用机制包括：降低心肌耗氧量,提高耐缺氧能力,清除自由基、抗氧化作用,调节血栓素A 2/前列环素,调节一氧化氮细胞内皮素1、抑制肿瘤坏死因子α和白细胞介素的释放等。 脐血间充质干细胞移植：移植成功最主要的障碍为移植后只有少量脐血间充质干细胞移植至心肌内并存活,因此提高脐血间充质干细胞向心肌内迁移和定植及其存活率对治疗效果具有重要意义。移植前对脐血间充质干细胞进行肿瘤坏死因子α预处理,发现肿瘤坏死因子α预处理胎儿脐血间充质干细胞移植治疗大白兔缺血性心脏病可以改善心脏功能、减少心肌梗死面积和心肌纤维化面积。 摘要 背景：心肌缺血损伤后,心肌细胞释放大量炎症递质作为对心肌损伤的应答,梗死及缺血区的炎症因子有助于心肌组织对损伤的修复和适应。 目的：探讨肿瘤坏死因子α预处理脐血间充质干细胞移植治疗对心肌梗死兔心功能的效果。 方法：将36只大白兔随机等分为4组,假手术组、模型组、无肿瘤坏死因子α组和肿瘤坏死因子α组,后3组建立心肌梗死模型。造模后24 h,模型组、无肿瘤坏死因子α组和肿瘤坏死因子α组分别在梗死中心区及边缘注射PBS、未经肿瘤坏死因子α预处理的脐血间充质干细胞及经肿瘤坏死因子α预处理的脐血间充质干细胞。 结果与结论：与模型组相比,肿瘤坏死因子α组和无肿瘤坏死因子α组兔心脏功能明显恢复,心肌梗死面积及心肌纤维化面积明显减小;且肿瘤坏死因子α组的效果优于无肿瘤坏死因子α组。表明肿瘤坏死因子α预处理胎儿脐血间充质干细胞移植能有效治疗心肌梗死。 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 ORCID: 0000-0002-4513-3726(王巍)     

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.     

骨髓间充质干细胞修复椎动脉型颈椎病的血管损伤

背景：炎症刺激是椎动脉型颈椎病发病的重要因素,骨髓间充质干细胞具有很强的免疫调节作用,能有效抑制炎症,具有治疗椎动脉型颈椎病的潜能。 目的：探讨椎动脉型颈椎病血管损伤机制及骨髓间充质干细胞的治疗作用。 方法：40只日本大耳兔随机分为4组：空白组不作任何处理;模型组、丹参酮组、干细胞组采用硬化剂注射法制作椎动脉型颈椎病模型。造模24 h后,空白组,模型组不进行干预,丹参酮组和干细胞组经耳缘静脉注射丹参酮ⅡA磺酸钠溶液10 mL和骨髓间充质干细胞混悬液10 mL,干预2周后取材。 结果与结论：与模型组相比,干细胞组管壁中膜平滑肌细胞肥大增生明显受到抑制,血管内皮皱折较匀称;干细胞组与丹参酮组比较无明显差别。与模型组相比,干细胞组椎动脉血管cathepsin B、白细胞介素6、肿瘤坏死因子α表达显著下降,差异有显著性意义(P < 0.05);模型组与丹参酮组比较差异无显著性意义(P > 0.05)。结果可见炎症反应可能是椎动脉血管受到损伤的机制之一;骨髓间充质干细胞能有效抑制椎动脉血管炎症反应,修复受损椎动脉血管。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：     

骨髓间充质干细胞移植治疗骨关节炎

BACKGROUND:As the ability of self-renewal, differentiation and migration into damaged tissues, bone marrow mesenchymal stem cells have been widely used in a variety of diseases. OBJECTIVE:To explore the therapeutic effect of bone marrow mesenchymal stem cell transplantation on osteoarthritis in rats. METHODS:Thirty-six Wistar rats were randomly assigned into transplantation, model or control group. osteoarthritis models were established in the transplantation and model groups, followed by tail vein injection of bone marrow mesenchymal stem cells (5×10⁷/kg) or the same volume of normal saline, respectively. Rats in the control group were subjected to no treatment. Four weeks after injection, levels of CD4⁺CD25⁺ regulatory T cells in the spleen, interleukin-17, tumor necrosis factor-α and transforming growth factor-β1 in serum were detected, and arthritis index and the degree of joint swelling were evaluated as well. RESULTS AND CONCLUSION:Compared with the control group, both arthritis index and degree of joint swelling were increased significantly in the model group (P < 0.05), but these indicators exhibited a remarkable improvement after cell transplantation (P < 0.05). Levels of CD4⁺CD25⁺ regulatory T cells in the spleen in the three groups were ranked as follows: the transplantation group > the control group > the model group. The levels of interleukin-17 and tumor necrosis factor-α in serum in the transplantation group were lower than those in the model group but higher than those in the control group (P < 0.05). Highest level of transforming growth factor-β1 was obtained in the transplantation group, followed by the control group and model group (P < 0.05). To conclude, these findings indicate that bone marrow mesenchymal stem cell transplantation exert therapeutic effects in osteoarthritis rats by immune and cytokine regulation.     

控释神经营养因子与细胞移植减少损伤脊髓的胶质瘢痕

背景：前期研究发现控释胶质细胞源性神经营养因子与骨髓间充质干细胞源神经元样细胞联合移植可有效促进猕猴脊髓损伤后运动功能和感觉功能的恢复。 目的：观察控释胶质细胞源性神经营养因子联合骨髓间充质干细胞源神经元样细胞移植抑制猴脊髓损伤后胶质瘢痕形成的作用是否优于单纯细胞移植。 方法：取12只恒河猴,采用改良Allen氏法制作急性重度脊髓损伤模型,随机数字表法分为3组,实验组以控释胶质细胞源性神经营养因子联合自体骨髓间充质干细胞源神经元样细胞移植修复,对照组以自体骨髓间充质干细胞源神经元样细胞移植修复,空白对照组以磷酸盐缓冲液修复。修复后5个月,取出脊髓组织制成石蜡标本,应用免疫组织化学染色显示胶质瘢痕的形态特征、构成特点及瘢痕中神经纤维的再生情况,检测胶质瘢痕面积及胶质纤维酸性蛋白染色的平均吸光度值。 结果与结论：脊髓损伤部位胶质瘢痕由混合性增生的星形胶质细胞和组织细胞构成。空白对照组脊髓胶质瘢痕累及范围广,星形胶质细胞增生显著,神经丝蛋白免疫组织化学染色阴性,胶质瘢痕面积、胶质纤维酸性蛋白染色平均吸光度值高于实验组与对照组(P < 0.05);实验组、对照组脊髓胶质瘢痕累及范围较局限,神经丝蛋白免疫组织化学染色显示有少量神经纤维通过瘢痕区,并且实验组胶质瘢痕面积、胶质纤维酸性蛋白染色平均吸光度值低于对照组(P < 0.05)。结果表明,控释胶质细胞源性神经营养因子联合骨髓间充质干细胞源性神经元样细胞移植可更强抑制脊髓损伤后胶质瘢痕的形成。     

乌司他丁联合脐带间充质干细胞移植脊髓损伤大鼠后肢运动功能和 运动诱发电位的变化

背景：乌司他丁能减轻炎性反应、清除氧自由基,对中枢神经系统损伤具有保护作用,能有效地提高脊髓损伤后移植细胞的存活率。 目的：观察乌司他丁联合脐带间充质干细胞移植对脊髓损伤大鼠后肢功能的影响。 方法：Wistar大鼠建立脊髓损伤动物模型后随机分成4组：空白对照组尾静脉注射培养液+腹腔注射生理盐水,细胞移植组尾静脉注射脐带间充质干细胞,乌司他丁组腹腔注入乌司他丁,联合组尾静脉注射脐带间充质干细胞,同时腹腔注入乌司他丁。 结果与结论：移植4周后联合组下肢运动功能优于细胞移植组和乌司他丁组(P < 0.05),细胞移植组和乌司他丁组优于空白对照组(P < 0.05)。移植后4周,PKH26标记的阳性细胞数联合移植组多于细胞移植组,细胞移植组多于乌司他丁组和空白对照组(P < 0.01)。移植后8周,联合组大鼠体感诱发电位及运动诱发电位的潜伏期、波幅明显优于其他3组(P < 0.05或P < 0.01)。提示乌司他丁联合应用脐带间充质干细胞移植可促进脊髓损伤大鼠神经突触的再生,改善其肢体运动功能和电生理功能,其效果优于单独应用乌司他丁或脐带间充质干细胞。     

脐血间充质干细胞移植修复急性肠缺血再灌注损伤

BACKGROUND:Bone marrow mesenchymal stem cells can repair intestinal ischemia-reperfusion injuny by interfering inflammatory reactions after intestinal ischemia-reperfusion to protect intestinal barrier functions. In recent years, umbilical cord blood mesenchymal stem cells are gradually used as a substitute source of bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the effects of umbilical cord blood mesenchymal stem cells on acute intestinal ischemia-reperfusion injury. METHODS:Umbilical cord blood mesenchymal stem cells were induced, isolated in vitro and tracked by CM-DiI fluorescent labeling. Sixty-three Sprague-Dawley rats were equivalently randomized into three groups: control group received normal saline enema, intestinal ischemia-reperfusion injury group with ethanol diluted trinitro-benzene-sulfonic acid and transplantation group administrated with 1×10¹⁰/L umbilical cord blood mesenchymal stem cell suspension via the tail vein at 1 hour after trinitro-benzene-sulfonic acid modeling. At 3 days after transplantation, colon tissues were removed in each group to observe pathological changes of the intestinal tract by hematoxylin-eosin staining. Besides, expression of leptin mRNA in the colon tissues and cyclooxygenase-2 in the mucosa were detected by RT-PCR and immunohistochemistry method, respectively. RESULTS AND CONCLUSION:Transplanted umbilical cord blood mesenchymal stem cells distributed in the intestinal lymphoid tissue and among glandular epithelial cells, suggesting that these stem cells might be involved in the process of intestinal ischemia-reperfusion injury repair. Compared with the control group, intestinal injury in the injury group was significantly aggravated, and most intestinal epithelial cells shed; and the transplantation group appeared to have significantly reduced intestinal damage and significantly less cell shedding. Expression of leptin mRNA was significantly higher in the injury group than the transplantation group followed by the control group, and there were significant differences among the three groups (P < 0.05). Additionally, expression of cyclooxygenase-2 in the injury group was significantly higher than that in the control group (P < 0.05); compared with the injury group, expression of cyclooxygenase-2 was significantly lower in the transplantation group (P < 0.05). To conclude, leptin and cyclooxygenase-2 may be involved in acute intestinal ischemia-reperfusion injury, and umbilical cord blood mesenchymal stem cell transplantation significantly lessens intestinal ischemia-reperfusion injury, which provides an experimental basis for human treating acute intestinal ischemic injury.     

骨髓间充质干细胞移植对胃癌模型大鼠肿瘤细胞的影响

BACKGROUND:Bone marrow mesenchymal stem cells can migrate into tumor tissues, as reported in recent studies. OBJECTIVE:To investigate the tropism and effect of transplantation of bone marrow mesenchymal stem cells on proliferation and differentiation of gastric cancer cell line SGC7901 in rats. METHODS:Gastric cancer models were established in Sprague-Dawley rats by subcutaneous injection of gastric cancer cell line SGC7901, and then, in cell transplantation group, each rat underwent an intraperitoneal injection of 1×107 bone marrow mesenchymal stem cells. After transplantation, the targeting ability of bone marrow mesenchymal stem cells was detected using fluorescent DiI labeling. Cyclin D2 mRNA and protein expressions were measured using real-time PCR and western blot assay, respectively. Apoptosis of gastric cancer cells was observed by in situ terminal labeling method. RESULTS AND METHODS: Bone marrow mesenchymal stem cells successfully migrated into the gastric cancer site in rats. The expression levels of cyclin D2 mRNA and protein in the cell transplantation group were significantly higher than those in the model group (P < 0.05). It was observed that the number of apoptotic cancer cells was significantly reduced in the cell transplantation group compared with the model group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells have the ability to migrate into the tumor sites, thereby promoting the proliferation of gastric cancer cells.     

骨髓间充质干细胞向脊髓损伤区的迁徙

背景：骨髓间充质干细胞移植到脊髓损伤区域后,如何观察其在体内的生存和转归情况,一直是让人困扰的问题。 目的：观察骨髓间充质干细胞在大鼠脊髓损伤区内的迁徙情况。 方法：36只Wistar大鼠随机分为2组,实验组制作脊髓损伤模型1周后,经尾静脉移植用DAPI标记的骨髓间充质干细胞(1×109 L-1) 1 mL,连续注射2 d。对照组未行脊髓损伤,与实验组同一时间同法行骨髓间充质干细胞移植。分别于移植后5,10,15 d,制作损伤脊髓冰冻切片,在激光共聚焦显微镜下观察骨髓间充质干细胞的迁徙情况。 结果与结论：实验组于移植后5 d,在脊髓损伤组织血管内出现少量荧光标记的骨髓间充质干细胞,10 d后有血管外弥散,15 d后有广泛弥散。对照组均未见DAPI标记的骨髓间充质干细胞。结果表明骨髓间充质干细胞经大鼠尾静脉移植后,能透过血脊髓屏障向损伤脊髓组织迁徙。