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1.
背景:骨髓间充质干细胞植入染矽尘大鼠体内能归巢到受损肺部,但何种植入途径在体内的归巢效果更好尚不清楚。
目的:动态比较观察不同途径移植骨髓间充质干细胞在染矽尘大鼠体内的分布情况。
方法:全骨髓贴壁法分离、培养供体大鼠骨髓间充质干细胞,用携带增强型绿色荧光蛋白基因的慢病毒(Lv-eGFP)转染骨髓间充质干细胞。受体大鼠用气管内注入法染尘,再随机分为经静脉组和经气管组,将转染Lv-eGFP的骨髓间充质干细胞分别经静脉、气管途径注入大鼠体内,在移植后的第1,2,3,4周处死大鼠,取心、肝、脾、肺、肾、脑组织进行冰冻切片,荧光显微镜下观察荧光,图文分析软件计算荧光强度。
结果与结论:两组大鼠肺组织均可见强烈、分布广泛且持续的绿色荧光,尤以气管、血管周围明显;两组荧光强度均随时间延长呈轻度减低趋势,但两组每周的荧光强度差异无显著性意义(P > 0.05)。两组大鼠其余各脏器早期也均可见荧光,其中肝、脾、心脏组织荧光强、分布广,肾、脑组织荧光相对较弱、分布较稀疏;随时间推移,各组织荧光均逐渐减弱,面积逐渐减少,到后期仅肝、脾组织可见到较弱、散在荧光分布,脑组织荧光几乎不可见。在同一时间点,两组除第1周脑组织荧光强度差异有显著性意义(P < 0.05)外,其余差异均无显著性意义(P > 0.05)。提示骨髓间充质干细胞经静脉和气管两种途径植入染矽尘大鼠体内归巢至受损肺部的效率相当。
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 相似文献
2.
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: 相似文献
3.
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。 相似文献
4.
背景:金属蛋白酶组织抑制因子1可能通过抑制基质金属蛋白酶活性降低干细胞侵袭能力,提高其归巢修复能力。
目的:观察转染金属蛋白酶组织抑制因子1基因骨髓间充质干细胞移植修复损伤心肌的能力。
方法:开胸结扎SD大鼠冠状动脉前降支建立心肌梗死模型,1周后随机分组:空白组经尾静脉内注射DMEM悬液;干细胞组经尾静脉内注射含1×107骨髓间充质干细胞的DMEM悬液;绿色荧光蛋白-干细胞组经尾静脉内注射转染带有绿色荧光蛋白基因慢病毒空载体的骨髓间充质干细胞(1×107)DMEM悬液;TIMP-1-shRNA-干细胞组经尾静脉内注射转染TIMP-1-shRNA的骨髓间充质干细胞(1×107)DMEM悬液。
结果与结论:造模后,4组大鼠心脏功能受损,收缩能力下降,治疗4周后心脏功能均有不同程度改善:与空白组比较,干细胞组、绿色荧光蛋白-干细胞组、TIMP-1-shRNA-干细胞组心脏收缩功能明显提高(P < 0.05),心肌梗死面积百分比明显缩小(P < 0.05),梗死区毛细血管密度明显增加(P < 0.05),且TIMP-1-shRNA-干细胞组改善程度优于干细胞组、绿色荧光蛋白-干细胞组(P < 0.05)。说明转染金属蛋白酶组织抑制因子1基因的骨髓间充质干细胞移植能够明显改善损伤心肌功能,修复缺血性心肌病。 相似文献
5.
背景:依达拉奉具有清除自由基和抑制脂质过氧化反应的作用,能够改善中枢神经系统损伤区的微环境。
目的:观察骨髓间充质干细胞移植联合依达拉奉治疗大鼠脑梗死的效果。
方法:采用线栓法建立大鼠大脑中动脉阻塞模型,随机分为3组,对照组经尾静脉注射细胞培养液,骨髓间充质干细胞组经尾静脉注射2.0×109 L-1的骨髓间充质干细胞悬液,依达拉奉+骨髓间充质干细胞组经尾静脉注射2.0×109 L-1骨髓间充质干细胞悬液同时经腹腔注射依达拉奉3 mg/(kg•d),连续5 d。移植后行神经功能缺损评分,应用RT-PCR测定脑梗死组织水通道蛋白9及水通道蛋白4 mRNA的表达,并经全脑冷冻切片苏木精-伊红染色及荧光显微镜观察细胞自然存活及分布情况。
结果与结论:移植后24 h,3 d各组间神经功能缺损评分差异无显著性意义(P > 0.05),移植后2周,依达拉奉+骨髓间充质干细胞组大鼠神经功能缺损评分低于骨髓间充质干细胞组及对照组(P < 0.05-0.01)。骨髓间充质干细胞组大鼠脑梗死周围组织水通道蛋白9 及水通道蛋白4 mRNA的表达高于依达拉奉+骨髓间充质干细胞组,却低于对照组(P < 0.05)。依达拉奉+骨髓间充质干细胞组CM-Dil阳性细胞和神经元数量多于骨髓间充质干细胞组及对照组(P < 0.05)。提示移植的骨髓间充质干细胞可移行至大鼠脑梗死灶周围并存活,分化为神经元样细胞。联合注射用依达拉奉治疗可明显改善脑梗死大鼠的神经学功能。 相似文献
6.
背景:研究miR-378对缺血缺氧条件下骨髓间充质干细胞生长增殖、细胞凋亡等生物学行为的影响,为进一步改善骨髓间充质干细胞在梗死心肌中的生存提供新方法。
目的:观察miR-378转染后的骨髓间充质干细胞耐受缺血缺氧及促进血管形成的能力。
方法:体外培养的骨髓间充质干细胞分为未转染组和转染组,未转染组为未转染miR-378的骨髓间充质干细胞,转染组则采用化学合成的miR-378模拟物转染SD大鼠骨髓间充质干细胞;将两组骨髓间充质干细胞置于缺血缺氧(无血清,体积分数为1%O2、5%CO2,94%N2)环境中培养24 h后,应用锥虫蓝染色计数法、MTS法检测两组细胞在不同时间点的生长及增殖情况,TUNEL法检测细胞凋亡情况;将两组缺血缺氧后的培养基上清液分别刺激人脐静脉内皮细胞,观察血管形成情况。
结果与结论:缺血缺氧干预后48 h和72 h,转染组的骨髓间充质干细胞活细胞数明显高于未转染组(均为P < 0.01);转染组的骨髓间充质干细胞表现出较高的增殖能力,缺血缺氧干预后24 h及48 h细胞增殖升高较未转染组明显,差异有显著性意义(P < 0.01,P < 0.05);缺血缺氧后转染组骨髓间充质干细胞的凋亡比例明显下降(P < 0.05)。两组细胞均可促进血管腔样结构形成,但转染组血管管腔样结构较未转染组显著增多(P < 0.01)。研究结果提示miR-378可促进缺血缺氧后骨髓间充质干细胞的生长增殖并抑制其在缺血缺氧条件下的细胞凋亡,同时可提高其促血管生成的能力。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: 相似文献
7.
背景:干细胞移植治疗肺动脉高压有一定疗效。
目的:观察骨髓间充质干细胞移植治疗肺动脉高压的效果及并探讨其作用机制。
方法:采用密度梯度离心法体外培养、纯化、扩增获得大鼠骨髓间充质干细胞,经荧光染料标记后备用。大鼠皮下注射野百合碱建立肺动脉高压模型,建模后1周将大鼠随机分为3组,干细胞移植组和肺动脉高压组大鼠皮下注射野百合碱建立肺动脉高压模型,1周后干细胞组大鼠经舌下静脉注射骨髓间充质干细胞悬液,肺动脉高压组注射等量不含干细胞的培养液,对照组皮下注射等量生理盐水。
结果与结论:移植后2周,与野百合碱诱导的肺动脉高压大鼠相比干细胞移植组血流动力学参数及右心室与体质量之比明显改善(P < 0.05);肺血管重构程度减轻(P < 0.05)。荧光显微镜下发现干细胞组移植的骨髓间充质干细胞在大鼠体内能存活至少2周,部分干细胞能转化为血管平滑肌细胞。说明静脉移植骨髓间充质干细胞能明显改善野百合碱造成的肺动脉高压大鼠肺血管和右心室结构的损伤。 相似文献
8.
背景:骨髓间充质干细胞分泌的血管内皮生长因子可能对心脏衰老有治疗作用,但这一作用是否与其能否归巢到心脏有关呢?
目的:分析骨髓间充质干细胞对衰老心脏的影响。
方法:取大鼠骨髓间充质干细胞。40只大鼠随机数字表法均分为正常组、造模组、治疗组和荧光标记剂组。后3组皮下注射D-半乳糖建立衰老心脏模型,后2组造模后尾静脉注射未标记和经羧基荧光素乙酰乙酸标记后的骨髓间充质干细胞;观察细胞是否定位于心脏、心脏组织各因子的表达。
结果与结论:骨髓间充质干细胞能够归巢到衰老心脏;与模型组比较,骨髓间充质干细胞治疗后能够使血清与心脏中超氧化物歧化酶活性均明显增高、丙二醛含量显著降低(P < 0.05)、心脏组织血管内皮生长因子表达升高(P < 0.05);说明骨髓间充质干细胞分泌血管内皮生长因子可能是其改善衰老心脏的机制之一。 相似文献
9.
背景:骨髓间充质干细胞具有分化成肾小管上皮细胞的功能,故推测,间充质干细胞可能具有治疗慢性肾功能减退的作用。
目的:研究骨髓来源的大鼠间充质干细胞防护肾脏衰老的作用及其可能的机制。
方法:实验建立衰老大鼠模型,尾静脉移植骨髓来源的大鼠间充质干细胞,应用荧光染料羟基荧光素二醋酸盐琥珀酰亚胺脂标记体外培养大鼠骨髓来源的间充质干细胞,将其回输肾脏衰老大鼠体内,应用荧光显微镜下观察间充质干细胞是否能够归巢于肾脏;全自动生化仪测定各组大鼠血清中尿素氮及肌酐水平变化;采用免疫荧光法,利用计算机图像叠加技术,观察蓝色荧光4’,6-二脒基-2-苯基吲哚分别与红色荧光的肾小管上皮细胞特异性蛋白-角蛋白的叠加情况。
结果与结论:骨髓来源的大鼠间充质干细胞能够归巢到衰老大鼠的肾脏。与衰老大鼠模型组相比,经过间充质干细胞生物治疗后的衰老大鼠血清中尿素氮及肌酐水平显著降低(P < 0.05),肾脏组织可见少量4’,6-二脒基-2-苯基吲哚及异硫氰酸荧光素标记的角蛋白双染阳性细胞。结果显示,骨髓来源的大鼠间充质干细胞通过向肾小管上皮分化,这可能是其改善衰老大鼠肾脏的机制之一。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: 相似文献
10.
黄永辉 《中国组织工程研究》2015,19(36):5811-5815
背景:骨髓间充质干细胞可以在体内外诱导下分泌神经营养因子,还可向神经元方向转化,从而促进创伤脑组织再生修复,但骨髓间充质干细胞又存在生存周期短的问题而影响其发挥对损伤脑组织的保护作用。目的:观察骨髓间充质干细胞移植联合神经节苷脂治疗大鼠颅脑创伤的效果。方法:60只Wistar大鼠采用液压颅脑损伤仪制备大鼠重型颅脑创伤模型,建模24 h后按照随机表法随机分为3组:模型组经尾静脉注入1 mL DMEM培养液,移植组经尾静脉注入1 mL(1×1010 L-1)骨髓间充质干细胞悬液,联合组经尾静脉注入1 mL(1×1010 L-1)骨髓间充质干细胞悬液以及经腹腔注射神经节苷脂水溶液(30 mg/kg),1次/d,共3 d。伤后24 h,细胞移植后3 d及1,2,3,4 周依据Longa 5分法进行神经功能行为学评分;移植后3 d应用RT-PCR、Western blot检测创伤脑组织中水通道蛋白4基因和蛋白的表达;移植后1周,行苏木精-伊红染色观察创伤脑组织病理变化。结果与结论:移植后3 d及1,2,3,4 周,联合组神经行为学功能评分低于移植组(P < 0.05),移植组低于模型组(P < 0.05);移植后3 d,模型组创伤脑周围组织水通道蛋白4蛋白及mRNA的表达高于移植组(P < 0.05),移植组高于联合组(P < 0.05);苏木精-伊红染色观察到联合组脑组织恢复明显优于模型组和移植组(P < 0.05)。结果提示骨髓间充质干细胞移植联合神经节苷脂可以明显改善颅脑创伤大鼠的神经行为学功能。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 相似文献
11.
背景:目前正在兴起的干细胞治疗技术及基因修饰技术有可能在肺动脉高压治疗中获得独特疗效。
目的:探讨携带碱性成纤维细胞生长因子基因的骨髓间充质干细胞移植对肺动脉高压大鼠血流动力学水平的影响。
方法:选择3周龄SD雄性大鼠,于体外进行骨髓间充质干细胞培养及纯化,在腺病毒介导下实施基因转染,使碱性成纤维细胞生长因子基因成功导入骨髓间充质干细胞。将60只SD大鼠给予野百合碱腹腔注射(50 mg/kg)进行肺动脉高压模型复制,随机分成3组,其中肺动脉高压组于颈内静脉移植1 mL的L-DMEM培养基,骨髓间充质干细胞组于颈内静脉移植1 mL空腺病毒载体转染的骨髓间充质干细胞悬液,碱性成纤维细胞生长因子组于颈内静脉移植1 mL腺病毒介导下碱性成纤维细胞生长因子转染骨髓间充质干细胞悬液。
结果与结论:经3周治疗后,各组大鼠动脉血压差异无显著性意义(P > 0.05);其中碱性成纤维细胞生长因子组大鼠的肺动脉收缩期压力和平均肺动脉压明显低于肺动脉高压组、骨髓间充质干细胞组,差异有显著性意义(P < 0.05);各组大鼠右心室的肥大指数比较差异无显著性意义(P > 0.05);碱性成纤维细胞生长因子组大鼠血浆内皮素1水平明显低于肺动脉高压组、骨髓间充质干细胞组,差异有显著性意义(P < 0.05)。结果显示携带碱性成纤维细胞生长因子的骨髓间充质干细胞移植,能够改善肺动脉高压大鼠肺血管的血流动力学水平,保护机体血管的内皮细胞,并拮抗血管的收缩。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: 相似文献
12.
赵海莲 《中国组织工程研究》2015,19(6):903-907
背景:人端粒酶反转录酶是调控增殖及定向分化的首选生长因子之一,具有多重生物学效应。
目的:观察人端粒酶反转录酶表达的骨髓间充质干细胞移植治疗大鼠糖尿病的效果。
方法:体外培养SD大鼠骨髓间充质干细胞,经反转录病毒PLXSN 为载体介导人端粒酶反转录酶基因转染骨髓间充质干细胞,在转染前后用RT-PCR检测骨髓间充质干细胞人端粒酶反转录酶基因的表达。60只雌性SD大鼠中随机取15只作为正常对照组,一次性注射生理盐水,余45只按45 mg/kg的剂量注射链脲霉素建立糖尿病模型后,随机分为3组,分别通过大鼠尾静脉注射移植人端粒酶反转录酶基因转染的骨髓间充质干细胞0.2 mL、骨髓间充质干细胞0.2 mL、生理盐水0.2 mL。
结果与结论:转染48 h后发现,骨髓间充质干细胞有人端粒酶反转录酶mRNA的表达,且重点集中于胞核内。移植后14 d,糖尿病组大鼠空腹血糖维持在较高水平,且高于正常对照组(P < 0.05);与糖尿病组相比,各移植组大鼠空腹血糖水平显著下降(P < 0.05);与骨髓间充质干细胞移植组相比,人端粒酶反转录酶基因转染的骨髓间充质干细胞移植组大鼠空腹血糖水平显著下降(P < 0.05),接近正常对照组水平(P > 0.05)。结果提示人端粒酶反转录酶表达的骨髓间充质干细胞移植能有效治疗大鼠糖尿病。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: 相似文献
13.
背景:血管内皮生长因子是一种有效的血管形成和通透性诱导因子,其中在体内以血管内皮生长因子165和血管内皮生长因子121表达为主,具有强烈的促血管新生作用。
目的:观察以血管内皮生长因子165基因转染骨髓间充质干细胞,继而分化为血管内皮细胞的可行性。
方法:分离提取50 g SD大鼠骨髓间充质干细胞,采用流式细胞仪鉴定,将携有血管内皮生长因子165基因的质粒pGLV-EF1a,采用慢病毒转染骨髓间充质干细胞,转染后于荧光显微镜下观察绿色荧光蛋白表达情况。
结果与结论:转染后12 h可见细胞内有绿色荧光蛋白表达,48 h 后表达增多,72 h后达到高峰,其后部分细胞荧光开始减退。结果证明血管内皮生长因子165基因转染骨髓间充质干细胞后,骨髓间充质干细胞内有绿色荧光蛋白表达,提示细胞转染成功,骨髓间充质干细胞定向分化为血管内皮细胞具有可行性。 相似文献
14.
BACKGROUND:Existing studies have shown that bone marrow mesenchymal stem cells can significantly improve islet function in diabetic rats to decrease excessively high blood glucose level, which may be related to the enhancement of differentiation ability of autologou pancreatic stem cells.
OBJECTIVE:To observe the therapeutic efficacy of basic fibroblast growth factor gene eukaryotic expression vector (PEGFP-C3-BFGF) transfection of bone marrow mesenchymal stem cells in diabetic rats.
METHODS:Recombinant adenovirus (Ad.aFGF) mediated PEGFP-C3-BFGF was transfected into bone marrow mesenchymal stem cells, and PEGFP-C3-BFGF expression was observed using fluorescence microscopy. Eighty Sprague-Dawley rats were randomly divided into normal control group, diabetes group, transplantation group, gene transfection group, with 20 rats in each group. After modeling, rats in different groups were given portal vein injection of normal saline, PBS, 1 mL of bone marrow mesenchymal stem cell suspension, and 1 mL of PEGFP-C3-BFGF-transfected bone marrow mesenchymal stem cell suspension. RT-PCR method was used to detect mRNA expression of matrix metalloproteinases in pancreatic tissue of rats in each group. Blood glucose levels of rats were detected at 24 hours, 3, 7, 14, 21 days after transplantation. ELISA method was used to detect plasma insulin levels in rats. Pathological changes of the pancreas were observed using hematoxylin-eosin staining.
RESULTS AND CONCLUSION:Under the fluorescence microscope, PEGFP-C3-BFGF transfected into cells after 48 hours showed significant specific red fluorescence. Two weeks after transplantation, matrix metalloproteinases mRNA expression was significantly increased in the diabetes group compared with the control group (P < 0.05), while it was decreased in the transplantation and gene transfection groups compared with the diabetes group (P < 0.05). After transplantation, the blood glucose levels in rats were ranked as follows: control group < gene transfection group < transplantation group < diabetes group (P < 0.05), and the plasma insulin levels in rats ranked as follows: control group > gene transfection group > transplantation group > diabetes group (P < 0.05). Pathological findings of the pancreas showed that the transplantation group was superior to the diabetes group, but inferior to the gene transfection group that was similar to the control group. All these findings indicate that PEGFP-C3-BFGF-transfected bone marrow mesenchymal stem cell transplantation can improve blood glucose levels and stimulate insulin secretion in diabetic rats, which may improve the severity of diabetes mellitus by decreasing the mRNA expression of matrix metalloproteinases. 相似文献
15.
BACKGROUND:Bone marrow mesenchymal stem cell transplantation for myocardial infarction becomes popularized in recent years, but transplanted cells cannot survive and proliferate under early inflammatory reaction or local ischemia/hypoxia microenvironment, eventually hampering the therapeutic outcomes.
OBJECTIVE:To investigate the therapeutic effect of PTEN-silenced bone marrow mesenchymal stem cells on acute myocardial infarction.
METHODS:(1) Bone marrow mesenchymal stem cells from Sprague-Dawley rats were randomly assigned to receive no treatment, NCsiRNA transfection using Lipofectamin2000 or PTEN siRNA transfection using Lipofectamin2000. Cell growth curves were described using MTT method to detect cell cycle using flow cytometry. (2) Thirty Sprague-Dawley rats were selected to prepare myocardial infarction models that were randomized into three groups (n=10 per group): blank control, negative control and RNAi group. Six hours after modeling, bone marrow mesenchymal stem cells transfected with nothing, NCsiRNA and PTEN siRNA were respectively injected into the infarcted center of the left ventricular anterior wall in these three rat groups. After 4 weeks, all rats were subjected to cardiac function detection using echocardiography, and the survival and proliferation of bone marrow mesenchymal stem cells in the rats were observed by fluorescence microscopy.
RESULTS AND CONCLUSION:Compared with the other two groups, a significant increase in the absorbance values at different culture time, the proportion of cells in S+G2 phase, and the number of bone marrow mesenchymal stem cells in the myocardial tissue was found in the RNAi group (all P < 0.05). Additionally, the left ventricular ejection fraction and left ventricular shortening fraction were significantly reduced in the RNAi group than the blank control and negative control groups at 4 weeks after cell transplantation (P < 0.05). Both in vivo and in vitro experimental findings showed that PTEN silencing could effectively improve cell survival and proliferation in the infarcted myocardium. Moreover, in the in vivo experiment, an overt improvement in rat’s cardiac function was achieved. 相似文献
16.
BACKGROUND:Bone marrow mesenchymal stem cell transplantation can effectively improve decreased cardiac function caused by heart failure, but there is a lack of research about the effect in bone marrow mesenchymal stem cell transplantation on cardiac function in heart failure induced by cardiomyopathies.
OBJECTIVE:To explore the effect of bone marrow mesenchymal stem cell transplantation on cardiac function in patients with cardiomyopathies accompanied by heart failure.
METHODS:Totally 40 Sprague-Dawley rats were enrolled, and bone marrow mesenchymal stem cells were isolated from 10 rats, and the remaining rats were equivalently randomized into normal, model and stem cell transplantation groups. Then rats in the model and stem cell transplantation groups were given intraperitoneal injection of hydrochloric acid doxorubicin to prepare cardiomyopathy-induced heart failure models. At 7 days after modeling, the stem cell transplantation group was treated with bone marrow mesenchymal stem cells through intravenous transplantation, and the model group was treated with equal amount of DMEM medium. Four weeks later, cardiac function of each rat was detected, and the cell survival and differentiation were observed by immunofluorescence method.
RESULTS AND CONCLUSION:At 4 weeks after transplantation, compared with normal and stem cell transplantation groups, the left ventricular systolic pressure and maximum rise/fall rate of left ventricular pressure were significantly decreased, but the left ventricular end diastolic pressure significantly increased in the model group (P < 0.05). And there was no significant difference between the normal group and the stem cell transplantation group (P > 0.05). High and dense fluorescence intensity was observed in the host myocardium immediately after transplantation. Subsequently, the fluorescence intensity and density decreased at 4 weeks, but the cell migration could be found, and some cells expressed cardiac troponin T. These results show that intravenous transplantation of bone marrow mesenchymal stem cells can improve cardiac function in rats with heart failure due to cardiomyopathies. Besides, the transplanted cells can survive in the host, and differentiate into cardiomyocyte-like cells. 相似文献
17.
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。 相似文献
18.
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. 相似文献
19.
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. 相似文献
20.
背景:骨形态发生蛋白2及转化生长因子β是骨再生中重要的因子,提高其表达可促进骨髓间充质干细胞的成骨分化。
目的:构建携带转化生长因子β3和骨形态发生蛋白2基因的慢病毒载体,观察其在骨髓间充质干细胞中的表达情况。
方法:应用重组慢病毒技术构建同时携带转化生长因子β3、骨形态发生蛋白2和绿色荧光蛋白基因的重组慢病毒表达载体,并用其转染体外培养的第3代兔骨髓间充质干细胞,以转染携带转化生长因子β3或骨形态发生蛋白2单一基因的慢病毒或单独慢病毒的骨髓间充质干细胞作为对照。转染后1周分别提取各组细胞的总RNA和蛋白进行检测。
结果与结论:荧光显微镜下见转染转化生长因子β3和(或)骨形态发生蛋白2基因3 d的骨髓间充质干细胞发绿色荧光,转染效率达90%以上。RT-PCR和Western blot结果显示,转染转化生长因子β3和骨形态发生蛋白2基因的骨髓间充质干细胞转化生长因子β3和骨形态发生蛋白2 mRNA和蛋白的表达均高于单一基因转染组及空白对照组。可见应用慢病毒可成功将转化生长因子β3和骨形态发生蛋白2基因转染至骨髓间充质干细胞并实现其高效表达,且两种基因具有协同促表达作用。 相似文献