干细胞生长因子和粒细胞集落刺激因子与心肌梗死大鼠体内骨髓间充质干细胞的迁移

背景：外周静脉移植间充质干细胞只有1%~5%的移植细胞能归巢到心肌梗死区域。 目的：观察干细胞生长因子、粒细胞集落刺激因子对骨髓间充质干细胞归巢的影响。 方法：采用贴壁培养法分离培养SD大鼠骨髓间充质干细胞,取传至3~5代细胞。建立SD大鼠急性心肌梗死模型,干细胞生长因子组、粒细胞集落刺激因子组、干细胞生长因子+粒细胞集落刺激因子组在骨髓间充质干细胞移植前3 d和移植后3 d单独或混合皮下注射干细胞生长因子、粒细胞集落刺激因子,骨髓间充质干细胞组不注射细胞因子。 结果与结论：荧光显微镜下观察,骨髓间充质干细胞迁移至心肌梗死组织,骨髓间充质干细胞组、干细胞生长因子组、粒细胞集落刺激因子组迁移至心肌梗死区的骨髓间充质干细胞数量没有明显的区别(P > 0.05),干细胞生长因子+粒细胞集落刺激因子组的骨髓间充质干细胞数量明显高于其他3组(P < 0.05)。免疫荧光组织化学显示,植入的部分骨髓间充质干细胞表达心肌特异蛋白cTnI。结果说明干细胞生长因子和粒细胞集落刺激因子两种细胞因子联合应用可以促进骨髓间充质干细胞归巢至心肌梗死区域,在体内微环境的诱导下,骨髓间充质干细胞能够转化为心肌样细胞。     

低氧增强骨髓间充质干细胞对缺氧诱导心肌细胞凋亡的可能性

背景：骨髓间充质干细胞移植入缺血心肌后存活率低,而低氧有可能增强骨髓间充质干细胞的增殖,促进其存活。 目的：体外模拟心肌细胞缺血微环境,探索低氧预处理后,骨髓间充质干细胞对持续缺氧诱导的心肌细胞凋亡的保护作用。 方法：取第4代SD大鼠骨髓间充质干细胞用于制备条件培养液。取胚胎大鼠心肌细胞株,随机分成4组：对照组：心肌细胞正常培养组;模型组：心肌细胞单纯缺氧;骨髓间充质干细胞组：心肌细胞与骨髓间充质干细胞条件培养液共缺氧;低氧组：心肌细胞与骨髓间充质干细胞低氧条件培养液共缺氧。MTT检测各组细胞活力变化,Annexin V-FITC双染标记心肌细胞凋亡,免疫组化检测各组Bax和Bcl-2蛋白的表达。 结果与结论：免疫组化显示,低氧组的Bcl-2表达较其他各组增强,而Bax的表达比模型组和骨髓间充质干细胞组减弱,Bcl-2/Bax比值最大。与对照组和骨髓间充质干细胞组相比,低氧组的细胞活力高(P < 0.05),凋亡率降低(P < 0.05)。提示低氧可能是通过增强旁分泌机制,从而对Bax和Bcl-2进行调节,对心肌细胞凋亡有保护效应。     

外科手术联合自体骨髓干细胞动员治疗大鼠脑缺血

背景：研究表明干细胞治疗能够促进脑功能恢复,联合外科干预能更有效地改善预后。 目的：分析外科手术联合自体骨髓干细胞动员治疗脑缺血的疗效和机制。 方法：用线栓法制作SD大鼠脑缺血模型后随机分为4组：对照组不干预,其他3组分别行外科手术、重组人粒细胞集落刺激因子和粒/巨噬细胞集落刺激因子动员自体骨髓干细胞治疗或联合治疗。 结果与结论：外科手术联合自体骨髓干细胞动员组大鼠的神经功能缺损评分、缺血坏死面积和细胞凋亡数明显低于对照组、外科手术组和自体骨髓干细胞动员组(P < 0.01);BrdU、BDNF阳性细胞数、微血管数较其他组明显增多(P < 0.05)。结果显示外科手术联合自体骨髓干细胞动员可减少大鼠脑梗死面积,抗细胞凋亡,促进神经再生和修复,改善脑功能。     

促红细胞生成素基因修饰骨髓间充质干细胞移植治疗大鼠脑梗死

背景：促红细胞生成素具有神经元的保护及促进神经再生的作用。目的：观察促红细胞生成素修饰的骨髓间充质干细胞尾静脉移植对大鼠脑梗死的治疗效果。方法：用Western blot鉴定外源人促红细胞生成素基因在骨髓间充质干细胞中的表达。采用线栓法建立大鼠大脑中动脉阻塞模型,模型组尾静脉注射PBS、骨髓间充质干细胞组注射骨髓间充质干细胞悬液,促红细胞生成素-骨髓间充质干细胞组注射转染了促红细胞生成素的骨髓间充质干细胞悬液。移植后3 d及移植后1,2,3,4 周行改良神经功能评分,检测神经功能的损伤情况。移植后4 周将大鼠麻醉后断头取脑,RT-PCR检测脑组织中bcl-2/bax基因表达变化,用原位末端标记法测定细胞凋亡情况、苏木精-伊红染色及荧光显微镜观察PKH26标记的骨髓间充质干细胞的存活和分布情况。结果与结论：Western blot结果显示,转染人促红细胞生成素基因的骨髓间充质干细胞体外能表达促红细胞生成素蛋白。移植后1-4周,骨髓间充质干细胞组和促红细胞生成素-骨髓间充质干细胞组神经缺损评分明显低于模型组(P < 0.05,P < 0.01)。与骨髓间充质干细胞组及模型组相比,大鼠脑梗死区组织促红细胞生成素-骨髓间充质干细胞组bcl-2基因的表达明显增高(P < 0.05),bax基因的表达明显降低(P < 0.05),凋亡细胞明显减少,PKH26阳性细胞数明显增多(P < 0.05)。结果证实,促红细胞生成素修饰的骨髓间充质干细胞尾静脉移植对脑梗死大鼠脑梗死疗效较好。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：     

沉默caspase-3基因影响骨髓间充质干细胞的增殖和凋亡

背景：缺血缺氧的心肌微环境导致植入的细胞存活率低。 目的：观察沉默caspase-3基因对大鼠骨髓间充质干细胞增殖和体外缺血缺氧环境下凋亡的影响。 方法：构建靶向caspase-3的shRNA重组慢病毒并转染骨髓间充质干细胞为转基因组,以正常细胞组和空载体组做对照,采用MTS法检测各组细胞增殖情况。建立缺血缺氧模型,real-time PCR和免疫组织化学分别检测缺血缺氧环境各组细胞的caspase-3 mRNA和蛋白表达水平,应用流式细胞术检测不同缺血缺氧时间点(0,6,12,24,48 h)各组细胞的凋亡率。 结果与结论：重组慢病毒成功转染骨髓间充质干细胞,且细胞增殖活性升高(P < 0.05)。缺血缺氧环境下,转基因组细胞caspase-3在mRNA和蛋白表达水平相比对照组下降(P < 0.05)。沉默caspase-3能显著降低骨髓间充质干细胞的凋亡率(P < 0.05),且随着缺血缺氧时间的延长凋亡率缓慢升高。结果提示,沉默caspase-3能加快骨髓间充质干细胞的生长速度和提高在体外缺血缺氧环境下的抗凋亡能力。     

肌肉注射脐带间充质干细胞对扩张型心肌病大鼠相关细胞因子的影响

背景：已有研究表明,在一定的剂量范围内,健康大鼠肌肉注射异种脐带间充质干细胞安全可靠,也证实了这一途径对扩张型心肌病大鼠心力衰竭同样安全有效。目的：探讨肌肉注射脐带间充质干细胞对扩张型心肌病大鼠相关细胞因子表达的影响。方法：雄性SD大鼠160只,随机抽取20只作为正常组,余大鼠通过阿霉素腹腔注射方式建立扩张型心肌病模型。造模成功大鼠随机分为模型组、上清液祖、低剂量组、中剂量组、高剂量组,分别经四肢肌肉注射人脐带间充质干细胞培养液、上清液及不同剂量的人脐带间充质干细胞,注射后观察大鼠一般情况,并于首次注射后4周再次给予相同剂量的培养液、上清液或脐带间充质干细胞。结果与结论：ELISA检测结果显示,肌注前后模型组大鼠血清肝细胞生长因子、白血病抑制因子、粒细胞集落刺激生物因子及血管内皮生长因子水平均较正常组增高(P < 0.05),肌注后低剂量组肝细胞生长因子、白血病抑制因子、血管内皮生长因子和粒细胞集落刺激生物因子较肌注前显著升高(P < 0.05),且较模型组明显升高(P < 0.05),中剂量组白血病抑制因子较肌注前明显升高(P < 0.05),高剂量组肝细胞生长因子、血管内皮生长因子和粒细胞集落刺激生物因子较肌注前无明显差异(P > 0.05)。免疫组化及RT-PCR结果均显示,各肌肉注射组大鼠心肌细胞胰岛素样生长因子1、血管内皮生长因子、肝细胞生长因子均较正常组表达增强,中剂量组呈强阳性表达,较其他各组增强明显。以上结果表明中、低剂量人脐带间充质干细胞肌肉注射可使扩张型心肌病大鼠血清肝细胞生长因子、白血病抑制因子、粒细胞集落刺激生物因子及血管内皮生长因子水平增高,并增加心肌组织胰岛素样生长因子1、肝细胞生长因子、血管内皮生长因子表达。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：     

人脐带间充质干细胞对异位子宫内膜细胞增殖及凋亡的影响

背景：间充质干细胞可以分泌多种细胞因子和生长因子,促进周围细胞的存活,发挥旁分泌作用。 目的：探讨人脐带间充质干细胞对异位子宫内膜细胞增殖及凋亡的影响。 方法：分离培养人脐带间充质干细胞和异位子宫内膜细胞,以异位子宫内膜细胞单独培养作为对照组,人脐带间充质干细胞与异位子宫内膜细胞共培养作为观察组。培养24,48,72 h时采用MTT法检测异位子宫内膜细胞增殖情况,流式细胞仪检测异位子宫内膜细胞凋亡情况,RT-PCR检测异位子宫内膜细胞PTEN基因表达情况。 结果与结论：与对照组比较,培养24,48,72 h时观察组异位子宫内膜细胞增殖受到显著抑制,亚二倍体峰占细胞总数的比例显著升高(P < 0.05)。随着时间的推移,观察组细胞抑制率逐渐下降,各时间点比较差异有显著性意义(P均 < 0.05)。与同期对照组比较,观察组细胞PTEN基因表达显著上调(P < 0.05)。培养48,72 h观察组细胞PTEN基因表达显著高于培养24 h (P < 0.05)。以上结果表明人脐带间充质干细胞可以通过上调PTEN基因表达,抑制异位子宫内膜细胞增殖,并促进其凋亡。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     

促红细胞生成素对急性肾损伤中干细胞因子及其受体表达的影响

背景：细胞因子在减轻肾脏缺血再灌注损伤中的作用日益受到重视,干细胞因子具有造血系统以外的器官保护作用。目的：探讨大鼠急性肾损伤模型中干细胞因子及其受体c-Kit表达的变化和促红细胞生成素预处理对其表达的影响。方法：成年雄性Wistar大鼠34只,采用夹闭双侧肾蒂建立缺血再灌注模型,缺血45 min后再灌注24 h,随机分为假手术组(n=10)、缺血再灌注组(n=12)和促红细胞生成素组(n=12),促红细胞生成素组于造模前2 h一次性尾静脉注射重组人促红细胞生成素5 000 U/kg。免疫组化及图像分析技术检测各组肾组织中干细胞因子及c-Kit的表达变化,测定血清肌酐和尿素氮水平,苏木精-伊红染色观察肾组织病理学改变并计算肾小管损伤积分。结果与结论：干细胞因子及c-Kit在肾组织中的表达仅限于肾小管区域。与假手术组比较,干细胞因子和c-Kit在缺血再灌注组和促红细胞生成素组的表达均明显增高(P < 0.05),促红细胞生成素组干细胞因子表达高于缺血再灌注组(P < 0.01),但两组c-Kit表达差异无显著性意义(P > 0.05);促红细胞生成素组血清肌酐与尿素氮水平明显低于缺血再灌注组(P < 0.05),但高于假手术组(P < 0.05)。与缺血再灌注组比较,促红细胞生成素组肾组织病变减轻。说明缺血再灌注导致急性肾损伤发生时干细胞因子及c-Kit表达升高,而促红细胞生成素对急性肾损伤的保护作用可能与上调干细胞因子及c-Kit表达有关。中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

干细胞因子和粒细胞集落刺激因子动员自身骨髓干细胞治疗缺血再灌注肾损伤

背景：骨髓干细胞可分化为肾脏组织固有细胞、修复损伤肾组织。正常情况下,外周血干细胞数目较少,骨髓干细胞动员剂可提高外周血干细胞数目。 目的：观察动员自身骨髓干细胞对缺血再灌注损伤肾脏修复作用及对缺氧诱导因子1α系统的影响,分析骨髓干细胞修复损伤肾脏的机制。 方法：SD大鼠随机分为4组。对照组不作处置;模型组制备肾缺血再灌注模型;治疗组给予缺血再灌注模型大鼠皮下注射骨髓干细胞动员剂干细胞因子200 μg/(kg•d)及粒细胞集落刺激因子50 μg/(kg•d),治疗对照组给予正常大鼠皮下注射与治疗组相同的药物,连续5 d。于术后5,10,17,24,31 d观察大鼠肾脏病理改变、骨髓干细胞表面抗原标记CD34⁺细胞表达以及缺氧诱导因子1α、血管内皮生长因子、血红素加氧酶1的表达变化。 结果与结论：①联合应用骨髓干细胞动员剂能明显增加损伤肾组织骨髓干细胞的数量,减轻肾组织损伤程度。②骨髓干细胞能促进肾组织缺氧诱导因子1α系统的表达,缺氧诱导因子1α系统及其靶基因产物血管内皮生长因子、血红素加氧酶1表达增加是骨髓干细胞促进急性肾损伤修复的可能机制之一。③骨髓干细胞动员剂对缺氧诱导因子1α系统的表达有一定的增强作用。     

重组人促红细胞生成素联合骨髓间充质干细胞移植治疗脓毒症大鼠心肌损伤

背景：相关研究表明促红细胞生成素及骨髓间充质干细胞均对心肌凋亡有一定影响,但两者联合应用治疗脓毒症相关性心肌损伤少见报道。 目的：观察骨髓间充质干细胞移植联合腹腔内注射促红细胞生成素对脓毒症大鼠心肌细胞病理学改变及细胞凋亡的影响。 方法：选用SD大鼠50只,随机数字表法均分为5组(n=10),应用盲肠结扎穿孔术建立脓毒症大鼠模型。骨髓间充质干细胞组建模后即刻尾静脉输注异体间充质干细胞;促红细胞生成素组建模后即刻腹腔内注射促红细胞生成素;促红细胞生成素+细胞移植组两者联合应用;模型组行盲肠结扎穿孔术,对照组开腹后不做任何处理,均尾静脉输注相同容量的生理盐水。24 h后麻醉处死实验动物,取心肌标本,采用苏木精-伊红染色观察心肌组织形态;用Western blot方法检测心肌组织中凋亡蛋白Bax、Caspase-3,抗凋亡蛋白Bcl-2的表达量。 结果与结论：苏木精-伊红染色结果：对照组可见心肌细胞排列整齐,心肌细胞结构完整;模型组可见广泛心肌纤维断裂,排列紊乱,心肌细胞肿胀或皱缩,可见空泡变性;心肌间质血管充血、水肿,炎症细胞浸润;促红细胞生成素组与骨髓间充质干细胞组心肌组织相似,炎性细胞浸润情况较轻,期间散在分布正常心肌细胞;促红细胞生成素+细胞移植组心肌细胞损害较轻,间质充血不明显,少量炎症细胞浸润。Western blot结果：促红细胞生成素+细胞移植组Bcl-2蛋白表达显著高于促红细胞生成素组、模型组及对照组(P < 0.01),Bax及Caspase-3蛋白表达量均减低(P < 0.05)。结果显示,骨髓间充质干细胞移植联合给予促红细胞生成素在脓毒症相关性心肌损伤的治疗中可减轻心肌的病理学改变、抑制心肌细胞的凋亡,其机制可能是通过上调抗凋亡蛋白、下调凋亡蛋白的表达来实现的。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：     

粒细胞集落刺激因子与血管性痴呆大鼠海马凋亡相关蛋白

背景：研究表明粒细胞集落刺激因子在保护神经元免受各种因素所致的神经元变性和死亡中发挥重要作用。 目的：观察粒细胞集落刺激因子对血管性痴呆大鼠海马组织神经细胞凋亡及Bcl-2、Bax蛋白表达的影响。 方法：采用永久性双侧颈总动脉结扎法建立SD大鼠血管性痴呆模型,以未进行血管结扎的大鼠作为假手术组。造模成功后,治疗组大鼠每日皮下注射粒细胞集落刺激因子50 μg/kg,假手术组和模型组注射等量的生理盐水。分别于造模后7,14,28 d取大鼠海马组织用于检测。 结果与结论：Morris水迷宫结果显示,模型组大鼠逃避潜伏期明显延长(P < 0.01),而治疗组各时间点大鼠逃避潜伏期较模型组缩短(P < 0.01);TUNEL及免疫组织化学结果显示,与模型组比较,治疗组各时间点大鼠海马TUNEL及Bax阳性细胞吸光度值明显减小(P < 0.01),Bcl-2阳性细胞吸光度值明显增加(P < 0.01)。说明粒细胞集落刺激因子可提高血管性痴呆大鼠海马Bcl-2蛋白的表达,抑制Bax蛋白的表达,减少神经细胞凋亡,改善大鼠的学习记忆功能。     

骨髓间充质干细胞移植联合依达拉奉抑制脑梗死后的神经元凋亡

背景：国内外多项研究证实骨髓间充质干细胞移植对脑梗死组织具有一定的神经保护作用。依达拉奉是一种新型强效小分子羟自由基清除剂,可通过清除脑梗死产生的自由基,抑制神经细胞损伤,从而起到脑保护作用。 目的：观察骨髓间充质干细胞移植联合依达拉奉对大鼠脑梗死组织水通道蛋白4、Bcl-2、脑源性神经营养因子表达的影响。 方法：选取Wistar大鼠80只,建立右侧大脑中动脉闭塞模型,随机分为对照组、骨髓间充质干细胞组、依达拉奉组和联合治疗组。建模6 h后通过尾静脉分别注入移植液,对照组注射培养液,骨髓间充质干细胞组注射骨髓间充质干细胞,依达拉奉组给予依达拉奉注射液,联合组同时注入骨髓间充质干细胞和依达拉奉注射液。分别在伤后72 h将大鼠麻醉后断头取脑,应用RT-PCR、Western Blot法检测脑组织中水通道蛋白4、Bcl-2、脑源性神经营养因子基因表达和蛋白合成变化。伤后12,24,36 h取大鼠脑组织以TUNEL法测定细胞凋亡情况。 结果与结论：RT-PCR、Western Blot结果显示,在骨髓间充质干细胞与依达拉奉联合治疗组中,Bcl-2、脑源性神经营养因子的表达明显高于骨髓间充质干细胞组、依达拉奉组及对照组(P < 0.05);而水通道蛋白4的表达低于其余各组(P < 0.05)。TUNEL测定结果显示,联合治疗组中免疫组化呈棕色的凋亡细胞明显少于单独治疗组及对照组。提示骨髓间充质干细胞移植与依达拉奉联合应用治疗大鼠脑梗死,可进一步促进损伤局部脑源性神经营养因子及Bcl-2的表达,对神经细胞凋亡具有明显的抑制作用,同时可下调水通道蛋白4水平,减轻脑水肿程度,二者联合运用的效果明显优于单独治疗组。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：     

维持性血液透析患者血清干细胞因子与皮肤瘙痒的关系

背景：目前慢性肾功能衰竭皮肤损害的发病机制尚未完全阐明。干细胞因子在慢性肾脏病及其并发症的发生、发展过程中可能起至关重要的作用。国内外有关维持性血液透析患者外周血干细胞因子水平的变化及其与皮肤瘙痒的关系鲜有报道。 目的：检测尿毒症维持性血液透析患者外周血干细胞因子水平的变化,并分析其与皮肤瘙痒的关系。 方法：采用ELISA法检测86例维持性血液透析患者外周血干细胞因子水平,采用目测类比评分法对尿毒症皮肤瘙痒程度进行评分,根据分值分为4组,0-2分组23例,3-5分组21例,6-8分组24例,>8分组18例。比较各组间干细胞因子水平的差异,并分析其与血红蛋白、全段血甲状旁腺激素的相关性。 结果与结论：各组患者性别、年龄、体质量指数和血压比较差异均无显著性意义(P > 0.05)。随着尿毒症皮肤瘙痒程度的加重,外周血干细胞因子水平逐渐升高(P < 0.05),血红蛋白水平逐渐降低(P < 0.05),全段血甲状旁腺激素逐渐升高(P< 0.05)。外周血干细胞因子水平与血红蛋白呈明显负相关(r =-0.60,P < 0.01);与全段血甲状旁腺激素呈明显正相关(r =0.7,P < 0.01)。提示外周血干细胞因子可能在尿毒症皮肤瘙痒的发生、发展过程中起重要作用。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：     

移植骨髓间充质干细胞肝癌大鼠细胞凋亡及炎症因子的动态变化

BACKGROUND:Bone marrow mesenchymal stem cell transplantation has not been thoroughly reported on its effects on apoptosis in hepatoma carcinoma cells and inflammatory factor level. OBJECTIVE:To investigate the effect of rat bone marrow mesenchymal stem cells on dynamic change of inflammatory factors and cell apoptosis during hepatocarcinogenesis. METHODS:Sixty healthy Sprague-Dawley rats were divided randomly into healthy group (n=30), control group (n=30) and transplantation group (n=30). Healthy group was given ordinary feed and normal water, while other groups were given diethylnitrosamine solution in drinking water to induce liver cancer models. Then, rats in the transplantation group were subjected to bone marrow mesenchymal stem cell transplantation via the tail vein. Two weeks after cell transplantation, CXCL5, interleukin-8 and interleukin-6 levels were tested by ELISA, mRNA level of hepatocyte nuclear factor 1α detected by RT-PCR, expression of Bcl-2 and Bax in liver tissue measured by immunohistochemical method, and liver cancer cell apoptosis index detected by TUNEL technique. RESULTS AND CONCLUSION:After modeling, the expressions of CXCL5, interleukin-8 and interleukin-6 in the control group were significantly higher than those in the healthy group (P < 0.05), while these indexes were reduced significantly after bone marrow mesenchymal stem cell transplantation (P < 0.05) and close to the normal levels (P > 0.05). Bone marrow mesenchymal stem cell transplantation significantly up-regulated the mRNA level of hepatocyte nuclear factor 1α in the liver tissue that was decreased obviously after modeling (P < 0.05). In addition, the expression of Bcl-2 was reduced, while the expression of Bax and the apoptosis index increased significantly in the transplantation group compared with the control group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cell transplantation contributes to hepatocyte differentiation and regeneration in liver cancer rats by reducing serum inflammatory factor levels and promoting apoptosis in hepatoma carcinoma cells.     

人羊膜间充质干细胞移植修复肝脏缺血-再灌注损伤

BACKGROUND:Studies have shown that human amniotic mesenchymal stem cells can differentiate into hepatocyte-like cells, suggesting that human amniotic mesenchymal stem cell transplantation provides a new potential for the clinical treatment of liver diseases. OBJECTIVE:To observe the effect of human amniotic mesenchymal stem cell transplantation on the repair of liver ischemia-reperfusion injury repair. METHODS:Sixty Sprague-Dawley rats were randomized into stem cell transplantation, model and control groups. Animal models of liver ischemia-reperfusion injury were made in the rats in the stem cell transplantation and model groups. One hour after modeling, rats in the stem cell transplantation were given injection of human amniotic mesenchymal stem cells (0.5 mL, 106 cells) via the tail vein, while rats in the model and control group were given L-DMEM (0.5 mL) or normal saline (0.5 mL), respectively. Liver function and liver morphology were detected at 1, 2, 3 weeks after transplantation. Meanwhile, RT-PCR detection and western blot assay were also conducted. RESULTS AND CONCLUSION:(1) Liver function: Compared with the control group, levels of aspartate aminotransferase, alanine aminotransferase and malondialdehyde were significantly increased in the model group at different time points after transplantation (P < 0.05), while a significant reduction in the levels of these three indicators was found after cell transplantation as compared with the model group (P < 0.05). (2) Liver morphology: 2 weeks after transplantation, rats in the model group exhibited hepatocyte degeneration and necrosis, and severe fibrosis, but these changes were remarkably alleviated in the stem cell transplantation group. (3) PT-PCR and western blot detection: 2 weeks after transplantation, a significantly higher level of hepatocyte growth factor in the liver tissue and a lower level of α-smooth muscle protein were found in the stem cell transplantation group compared with the model group (P < 0.05). All these experimental findings indicate that human amniotic mesenchymal stem cell transplantation can improve impaired liver function in rats, possibly through regulating hepatocyte growth factor and α-smooth muscle protein expression levels in the liver, and thereby promotes the repair of liver ischemia-reperfusion injury.     

视网膜干细胞移植对青光眼视网膜神经节细胞的保护

BACKGROUND:Stem cell transplantation is a new method for blinding eye disease. But there is a lack of research about the protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma. OBJECTIVE:To explore the protective effect of retinal stem cell transplantation on retinal ganglion cells of rats with glaucoma. METHODS:Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group) including control, model and retinal stem cell transplantation groups. Rat models of glaucoma were prepared in the latter two groups, and at 7 days after modeling, rats in the three groups were given intravitreal injection of 1 mL retinal stem cells (5x10⁶ cells), the same amount of PBS, and no treatment, respectively. Subsequently, relative indicators were detected at 2 weeks after transplantation. RESULTS AND CONCLUSION:The expressions of brain-derived neurotrophic factor and insulin-like growth factor I protein as well as the number of retinal ganglion cells were the highest in the control group, followed by the retinal stem cell transplantation group model group, and the lowest in the model group (P < 0.05). The number of apoptotic retinal ganglion cells in model group was significantly higher than that of control group (P < 0.05), and which in the retinal stem cell transplantation group was significantly lower than that in the model group (P < 0.05), but higher than that in the control group (P < 0.05). These results suggest that retinal stem cell transplantation for rat glaucoma can exert a protective effect on retinal ganglion cells.     

骨髓间充质干细胞移植脑梗死大鼠：神经功能恢复与突触素的表达

BACKGROUND:Synaptophysin plays an important role in the recovery of neural function after cerebral ischemia. OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem cell transplantation on nervous function and expression of synaptophysin after cerebral infarction. METHODS:Totally 60 rats were equivalently randomized into four groups, including sham operation, control, model and stem cell treatment groups. Rats in the control, model and stem cell treatment groups were used for preparing cerebral infarction models, and the remaining underwent the sham operation. After 1 day of modeling, bone marrow mesenchymal stem cells were transplanted into the rat lateral ventricle in the stem cell treatment group, and rats in the control group was given the injection of the same amount of PBS. After 1, 7 and 14 days of treatment, rat’s neurological function was scored on beam-walking test, rotarod test and screen test, and expression of synaptophysin was detected by RT-PCR and immunohistochemical assay. RESULTS AND CONCLUSION:At 7 and 14 days after treatment, the beam-walking test, rotarod test and screen test scores in the stem cell treatment group were significantly lower than those in the control and model groups (P < 0.05), and the above scores showed no significant differences between the control group and model group (P > 0.05). At 1 day after treatment, the mRNA expression of synaptophysin and the number of synaptophysin-positive cells in the sham operation group were significantly higher than those in the other three groups (P < 0.05); at 7 and 14 days after treatment, the mRNA expression of synaptophysin and the number of synaptophysin-positive cells in the stem cell treatment group were significantly increased compared with the other three groups (P < 0.05), and additionally, the mRNA expression of synaptophysin and the number of synaptophysin-positive cells in the sham operation group were significantly lower than those in the model and control groups (P < 0.05). These findings suggest that bone marrow mesenchymal stem cell transplantation can effectively promote the recovery of neurological function in cerebral infarction rats, and partially promote the formation of synaptophysin.     

红景天苷与淤胆血清诱导骨髓间充质干细胞向肝样细胞分化

背景：大量实验证实骨髓间充质干细胞在诱导因子及特定微环境下可诱导分化为肝细胞,并已广泛用于终末肝病的临床替代治疗,而其最佳诱导条件目前尚不清楚。 目的：初步探讨中草药红景天苷联合淤胆大鼠血清体外诱导大鼠骨髓间充质干细胞向肝细胞分化的可行性和有效性。　 方法：采用全骨髓贴壁培养法从大鼠骨髓中获取骨髓间充质干细胞,流式法检测干细胞表型;胆总管结扎切断法制备大鼠淤胆血清。取第3代骨髓间充质干细胞分3组体外诱导培养：空白对照组,基础培养基+5%淤胆血清;红景天苷组：基础培养基+5%淤胆血清+30 µmol/L红景天苷;阳性对照组：基础培养基+5%淤胆血清+20 µg/L肝细胞生长因子;观察各组诱导培养过程中细胞形态变化,RT-PCR法、Western-Blot法检测各诱导组肝细胞特异性蛋白表达水平。 结果与结论：骨髓间充质干细胞高表达CD90、CD105,不表达CD45、CD14、CD34、CD79a;空白对照组、红景天苷组、阳性对照组细胞在诱导培养中均出现多角及双核细胞;空白对照组、红景天苷组、阳性对照组在诱导培养7 d开始出现甲胎蛋白、白蛋白的mRNA及蛋白表达;在同一时间点空白对照组表达率最低(P < 0.05),红景天苷组、阳性对照组间比较,差异无显著性意义(P > 0.05)。与传统淤胆血清体外诱导相比,红景天苷联合淤胆血清能更有效诱导骨髓间充质干细胞向肝样细胞分化。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：     

黄芪甲苷孵育脂肪源性干细胞对顺铂诱导肾小管上皮细胞凋亡的影响

背景：干细胞移植用于治疗急性肾损伤的有效性已经被多个研究证实,但其对肾小管上皮细胞损伤的修复机制尚不明确。 目的：观察黄芪甲苷孵育后的脂肪源性干细胞对顺铂诱导的肾小管上皮细胞凋亡的保护作用及机制。 方法：实验分为4组。2.5 μmol/L顺铂诱导肾小管上皮细胞 24 h,建立肾小管细胞损伤模型(顺铂损伤组);将脂肪源性干细胞与损伤肾小管上皮细胞共培养(脂肪源性干细胞+损伤肾小管上皮细胞组);利用Transwell小室将20 mg/L黄芪甲苷孵育脂肪源性干细胞48 h后与损伤肾小管上皮细胞共培养(黄芪甲苷孵育脂肪源性干细胞+损伤肾小管上皮细胞组);以正常肾小管上皮细胞做对照(正常对照组)。 结果与结论：与肾小管上皮细胞损伤组相比,AV/PI和TUNEL结果均显示脂肪源性干细胞+肾小管上皮细胞组和20 mg/L 黄芪甲苷脂肪源性干细胞+肾小管上皮细胞组肾小管上皮细胞发生凋亡的比例和数量明显减少;ELISA结果表明20 mg/L黄芪甲苷脂肪源性干细胞+肾小管上皮细胞组胰岛素样生长因子1分泌显著提高(P < 0.05);Western blot进一步显示20 mg/L 黄芪甲苷脂肪源性干细胞+肾小管上皮细胞组caspase-3蛋白水平明显下降,而Bcl-2的表达量明显增加(P < 0.05)。表明黄芪甲苷孵育的人脂肪源性干细胞对顺铂诱导的肾小管上皮细胞凋亡具有抑制作用,从而有利于肾小管损伤的早期恢复,其保护机制可能与增加胰岛素样生长因子1分泌,抑制caspase-3表达、上调Bcl-2水平有关。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：     

碱性成纤维细胞生长因子基因修饰骨髓间充质干细胞移植修复急性肾损伤

BACKGROUND:It has been confirmed that basic fibroblast growth factor (bFGF) can promote the growth, proliferation, differentiation and functional expression of most cells derived from neuroderm and mesoderm. OBJECTIVE:To investigate the effect of bFGF-transfected bone marrow mesenchymal stem cell transplantation in rats with acute kidney injury. METHODS:bFGF genes were transfected into bone marrow mesenchymal stem cells via an adenovirus vector, and then expression of bFGF in transfected cells was identified using RT-PCR technology. Rat models of acute kidney injury were prepared by clipping bilateral renal pedicles, and then randomized into three groups (n=20): rats were given injection of bone marrow mesenchymal stem cell suspensions via tail vein as negative transfected group, those given injection of bFGF-transfected bone marrow mesenchymal stem cell suspensions via tail vein as bFGF-transfected group, and the others given injection of DMEM via tail vein as model group. Four weeks later, levels of serum creatinine and urea nitrogen were detected, expressions of connective tissue growth factor and growth factor in renal tissues were detected by Western blot assay, and morphology of renal tissues was observed using hematoxylin-eosin staining. RESULTS AND CONCLUSION:bFGF genes were successfully transfected into bone marrow mesenchymal stem cells. Compared with the model group, the levels of serum creatinine and urea nitrogen were significantly reduced in bFGF-transfected and negative transfected groups, especially in the bFGF-transfected group (P < 0.05), while expressions of connective tissue growth factor and transforming growth factor in renal tissues in bFGF-transfected and negative transfected groups were significantly weakened in these two groups (P < 0.05), but there were no significant differences between the bFGF-transfected group and negative transfected group (P > 0.05). Besides, renal tissues necrosis and inflammatory reactions were mitigated in the negative transfected group; renal tubules with normal outlines and no overt necrotic cells could be found in the bFGF-transfected group. These findings show that bFGF-transfected bone marrow mesenchymal stem cell transplantation plays a better role in acute kidney injury repair in rats.