间充质干细胞治疗皮瓣缺血再灌注损伤的作用机制及优势

背景：间充质干细胞因具有抗氧化、抑制炎症反应和诱导血管新生等作用而被用于皮瓣缺血再灌注损伤中。目的：综述间充质干细胞在治疗皮瓣缺血再灌注损伤方面的作用机制和最新进展,为其进一步理论研究和临床合理应用提供依据。方法：检索CNKI中国期刊全文数据库、万方数据库和PubMed数据库收录的相关文献。中文检索词为“间充质干细胞;皮瓣缺血再灌注损伤;条件培养基;外泌体;氧化应激;炎症反应;血管新生”。英文检索词为“mesenchymal stem cells;flap ischemia reperfusion injury;Conditioned medium;exosomes;Oxidative stress;Inflammatory reaction;Angiogenesis”。检索2010年以来相关文献,通过阅读文章剔除研究内容与文章主题关系不大、质量较差及内容陈旧文献,最终纳入74篇文献进行归纳总结。结果与结论：(1)间充质干细胞在抗氧化、抑制炎性反应及诱导血管新生等方面作用显著,在治疗皮瓣缺血再灌注损伤中蕴含着巨大潜力。(2)然而,间充质干细胞自身的缺陷及近年来治疗效果的下降使间充质干细...     

骨髓间充质干细胞移植促进大鼠肾缺血再灌注损伤的恢复

目的 观察骨髓间充质干细胞(MSCs) 缺血再灌注肾损伤大鼠体内的分化情况及对肾修复的促进作用.方法 72只雌性6周龄sD大鼠随机分为正常组、假手术组、缺血再灌注(I/R)组、MSCs移植组4组,分别于再灌注后12 h、3、7、14、42 d随机选取I/R组和MSCs组大鼠各6只,收获肾脏标本和血标本.测定血标本尿素氮和肌酐值;肾脏切片行HE染色、免疫组化PCNA染色、TUNEL法检测原位凋亡、激光共聚焦显微镜观察MSCs分化情况.结果 再灌注后7 d内,与I/R组比较,MSCs组BUN值、Scr值明显降低(P<0.05),组织学评分明显减低,PCNA阳性细胞数明显增多(P<0.05).再灌注后12 h,MSCs组凋亡细胞数少于I/R组(P<0.05).再灌注后42 d,肾小管中有BrdU阳性细胞.结论 MSCs移植可减轻急性肾缺血再灌注损伤鼠肾小管上皮细胞的损伤,促进肾功能恢复.少部分MSCs在体内可分化形成肾小管上皮细胞.     

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.     

干细胞移植与肺缺血再灌注损伤

肺移植是治疗终末期肺疾病的唯一有效手段,但目前全球对于肺移植的治疗效果仍不能达到理想的要求,其中主要因素是供体肺组织由于缺血再灌注而导致的器官损伤,从而影响了移植物功能,但目前尚无有效的治疗措施,国际上对于肺缺血再灌注的治疗多以常规手段为主,但治疗效果难以突破完全修复损伤的技术瓶颈.将干细胞治疗引入到缺血再灌注损伤的治疗当中会带来新的希望,明确干细胞中的各个细胞亚群所起到的作用及其间的相互协调及辅助作用,会为临床治疗找到新的方向.     

骨髓间充质干细胞移植保护小肠缺血再灌注损伤

BACKGROUND: Bone marrow mesenchymal stem cells have good proliferation and paracrine functions, which have irreplaceable advantages in the treatment of intestinal diseases. OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cell transplantation on intestinal ischemia-reperfusion injury in rats. METHODS: Forty-eight Sprague-Dawley rats were enrolled to make animal models of ischemic reperfusion injury of the intestine, and then model rats were randomized into experimental and control groups. After modeling, 1 mL bone marrow mesenchymal stem cells or the same volume of normal saline were injected into the intestinal mucosa of rats in the two groups, respectively. At hours 0, 2, 6, 24, 72, 120 after injection, serum diamine oxidase, tumor necrosis factor α, and D-lactic acid levels were detected by ELISA method. At 24 hours after injection, rat intestinal tissues were taken and observed pathologically under light microscopy, and their close connections were observed under transmission electron microscope. ZO-1 protein levels were detected by immunohistochemistry method. RESULTS AND CONCLUSION: Compared with the control group, the serum diamine oxidase, tumor necrosis factor α, and D-lactic acid levels were significantly lower in the experimental group at hours 6 and 24 after injection (P < 0.05). Intestinal necrosis, villous edema, intestinal congestion and inflammatory cell infiltration in the experimental group were milder than those in the control group. In addition, the ZO-1 protein expression in the experimental group was higher than that in the control group. Experimental results show that bone marrow mesenchymal stem cell transplantation into the intestinal mucosa can improve the intestinal mucosal permeability in rats with intestinal ischemia-reperfusion injury.      

皮瓣缺血再灌注损伤治疗研究进展

在皮瓣移植术后常常会发生缺血再灌注损伤,缺血再灌注损伤是引起皮瓣部分或全部坏死的重要原因.目前对皮瓣缺血再灌注损伤的发病机制尚不清楚,大多数学者认为可能与细胞凋亡、炎性反应、组织微循环障碍、细胞氧化损伤等有关.学者们提出用缺血预处理,药物干预,减少炎性反应,移植干细胞或干细胞联合基因疗法,体外冲击波,抗氧化反应和抑制细...     

骨髓间充质干细胞移植促肾缺血-再灌注损伤修复的机制初探

目的: 探讨骨髓间充质干细胞(BMSCs)促肾缺血-再灌注损伤修复的能力及机制.方法: 取健康3周龄C57BL/6J小鼠的骨髓细胞悬液,进行BMSCs体外扩增培养.将8周龄C57BL/6J小鼠随机分为假手术组(10只)、平行对照组(10只)和细胞移植组(28只).建立肾缺血-再灌注损伤模型,将BMSCs移植到细胞移植组小鼠中,平行对照组注射生理盐水.通过检测肾功能观察BMSCs对肾损伤的促修复作用.并通过检测氧化应激指标和观察蓝色荧光标记供体BMSCs在受体内的踪迹,初步探讨BMSCs促宿主损伤肾组织修复的机制.结果: 分离培养的BMSCs增殖旺盛,纯度较高,且均质性和稳定性好.平行对照组表现为小鼠整体状态差,肾功能指标和氧化应激指标都没有得到改观.细胞移植组小鼠整体状态良好,肾功能指标和氧化应激指标明显改善(P<0.05).并在宿主恢复的肾组织中发现了较多的带有蓝色荧光的细胞存在.结论: BMSCs在体外易分离培养,具有旺盛的增殖能力;移植BMSCs后,具有向受损组织归巢的作用,并参与和提高肾缺血-再灌注损伤后宿主C57BL/6J小鼠的恢复.     

胎盘间充质干细胞对小鼠小肠缺血再灌注损伤的作用

目的探讨胎盘间充质干细胞(pMSCs)对小鼠小肠缺血再灌注损伤的作用。方法组织块培养法提取pMSCs,流式细胞技术鉴定。24只小鼠随机分为假手术组(Sham组),缺血30min再灌注6h组(I/R组)和再灌注1h尾静脉输入5×10 6个pMSCs组。缺血再灌注6h后观察肠道情况,HE染色观察肠组织形态,ELISA检测血清中二胺氧化酶(DAO)、TNF-α、IL-6及IL-10含量。结果 pM SCs阳性表达CD29,CD44。与Sham组相比,I/R组肠道损伤明显,肠黏膜上皮细胞脱落多,而pM SCs组肠道损伤轻,细胞脱落少。I/R组和pM SCs组小鼠血清中DAO、TNF-α、IL-6及IL-10含量明显多于Sham组(P0.05)。与I/R组相比,pM SCs组小鼠血清中DAO、TNF-α、IL-6含量减少(P0.05),IL-10含量增多(P0.05)。结论 pM SCs可减轻小鼠小肠缺血再灌注损伤,机制与抑制再灌注后炎症反应相关。     

骨髓间充质干细胞治疗肾缺血再灌注损伤的旁分泌机制

背景：骨髓间充质干细胞已用于肾缺血再灌注损伤修复的实验动物研究。 目的：探讨骨髓间充质干细胞治疗肾缺血再灌注损伤的旁分泌机制。 方法：体外培养、纯化并体外DAPI标记大鼠骨髓间充质干细胞,经下腔静脉移植入肾缺血再灌注损伤模型大鼠体内,观察骨髓间充质干细胞对肾缺血再灌注损伤肾功能的保护作用以及在受体鼠体内的迁移情况,并应用免疫组织化学法检测骨髓间充质干细胞治疗后第2天缺血肾脏组织中血管内皮生长因子、肿瘤坏死因子α细胞因子的表达。 结果与结论：与注射生理盐水的对照组比较,细胞移植组大鼠血清肌酐和尿素氮水平在移植后第1、2天明显降低(P < 0.05),但细胞移植组移植后第1、2天肾组织中均未见DAPI阳性细胞;第3、4天则逐渐可见DAPI阳性细胞。免疫组织化学染色结果显示,与对照组相比,移植后第2天肾组织中可见较多血管内皮生长因子阳性细胞,而肾组织中肿瘤坏死因子α阳性细胞明显减少。结果显示旁分泌机制参与了骨髓间充质干细胞治疗肾缺血再灌注损伤。     

缺氧预处理人华通胶间充质干细胞条件上清对心肌缺血再灌注损伤的作用及机制

背景:间充质干细胞主要通过旁分泌作用改善组织损伤,低氧预处理后其旁分泌作用增强,但关于人华通胶间充质干细胞低氧预处理后的条件上清液对心肌缺血再灌注损伤的作用及其机制仍需深入研究.目的:探讨缺氧预处理人华通胶间充质干细胞条件上清对心肌缺血再灌注损伤H9C2细胞凋亡的影响及机制.方法:常氧、低氧处理人华通胶间充质干细胞,获...     

己酮可可碱缓解大鼠皮瓣缺血再灌注损伤

近年来有陆续报道己酮可可碱用于脑[1]、肺、肝等脏器的缺血再灌注损伤,但对于皮瓣缺血再灌注的作用未见报道;本实验拟明确其对皮瓣缺血再灌注损伤的保护作用,为减轻皮瓣缺血再灌注损伤、提高皮瓣移植成功率提供一定的实验依据.     

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.     

尾静脉注射脂肪间充质干细胞修复大鼠脑缺血再灌注损伤

目的探讨经尾静脉注射脂肪间充质干细胞是否对脑缺血再灌注损伤有一定的修复作用及相关机制。方法 45只SD大鼠随机均分为假手术组(sham),模型组(I/R)和治疗组(ADMSCs)。I/R和ADMSCs组大鼠采用线栓法大脑中动脉脑缺血再灌模型(MCAO)建立局灶性脑缺血再灌动物模型。缺血2h后再灌注,6ADMSCs组再灌注0和12h经尾静脉注射2.0×10个(0.5ml)脂肪间充质干细胞,I/R组注射等量的生理盐水。再灌注24h后处死大鼠并迅速取脑组织。通过TTC染色计算脑梗塞面积;TUNEL检测缺血部分脑组织中的细胞凋亡情况;Western blot检测缺血部分脑组织中i NOS和bcl-2/bax的表达水平。结果 ADMSCs组显著降低了脑梗塞体积,抑制神经细胞凋亡,减少Bax/bcl-2蛋白比,i NOS的表达水平明显下调。结论尾静脉注射ADMSCs修复脑I/R损伤可能与抑制神经细胞凋亡和i NOS表达相关。     

两种示踪方式的骨髓间充质干细胞在促进肝缺血再灌注损伤修复中的对比研究

目的　比较慢病毒转染绿色荧光蛋白(GFP)的骨髓间充质干细胞(BMSCs)和GFP转基因的BMSCs在大鼠肝缺血再灌注损伤修复中修复时效性及荧光稳定性的差异。 方法　常规体外培养2种BMSCs,MTT法检测二种细胞生长曲线间的异同;将40只SD大鼠随机分为假手术组、模型组、慢病毒转染GFP组(LV-GFP组)和GFP转基因组(GFP-BMSCs组),造模后LV-GFP组及GFP-BMSCs组于门静脉立即注入相应细胞悬液200 μl (数量约1×106个),模型组注入等体积的PBS溶液。于术后1、2、3、4周检测4组大鼠天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)及血清白蛋白(ALB)水平;于术后1~5 d切取实验组肝脏组织检测BMSCs入肝情况;于术后4周切取实验组肝脏组织检测BMSCs的荧光稳定性及其肝角蛋白18(CK18)的表达情况。 结果　GFP转基因大鼠的BMSCs在对数期的增殖能力明显强于慢病毒转染GFP的BMSCs(P<0.05);术后1、2周 GFP-BMSCs组AST及ALT水平明显低于 LV-GFP组 (P<0.05),术后2、3周 GFP-BMSCs组ALB水平明显高于 LV-GFP组(P<0.05);GFP-BMSCs组与LV-GFP组分别于术后3、5 d在肝区内见到GFP标记的BMSCs细胞;GFP-BMSCs及LV-GFP组都可于术后4周在肝区内见到已分化为肝细胞的BMSCs细胞,但GFP-BMSCs组BMSCs的荧光强度明显优于LV-GFP组。 结论　GFP转基因大鼠的BMSCs在大鼠肝缺血再灌注损伤修复中较慢病毒转染GFP的BMSCs展现出较好的修复时效性及荧光稳定性。     

异丙酚预处理联合脐血间充质干细胞移植治疗脑缺血再灌注损伤

BACKGROUND:As propofol has a neuroprotective effect, and umbilical cord blood mesenchymal stem cells have a high differentiation potential, their combination will have a better therapeutic effect on cerebral ischemia-reperfusion injury. OBJECTIVE:To study the effect of propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation on cerebral ischemia-reperfusion injury in rats. METHODS:Sixty-three Sprague-Dawley rats were randomized into model, propofol, and combined group (n=21 per group). Rat models of cerebral ischemia-reperfusion injury were made using ligation of the middle cerebral artery occlusion in the three groups. Rats in the combined group were given 100 mg/kg propofol injection at 1 day before injury and injection of umbilical blood mesenchymal stem cells via the tail vein (0.5 mL, 2×10⁹/L). RESULTS AND CONCLUSION:Compared with the model group, the neurological function was improved significantly in the propofol and combined group, especially in the latter one, presenting with a remarkable mitigation in brain injury and an increased level of survivn mRNA in the rat hippocampus. The content of serum malondialdehyde was lower but the activity of superoxide dismutase was higher in the combined group compared with the propofol group. These findings indicate that propofol pretreatment combined with umbilical blood mesenchymal stem cell transplantation has better therapeutic effects than propofol pretreamtnet alone for improving cerebral ischemia-reperfusion injury in rats.     

低氧预处理骨髓间充质干细胞减轻脊髓缺血再灌注损伤的作用机制

脊髓缺血再灌注损伤是脊柱减压手术后常见的并发症,骨髓间充质干细胞椎管鞘内移植为治疗此损伤的有效方法之一。目前许多研究表明低氧预处理可促进骨髓间充质干细胞功能并减轻脊髓缺血再灌注损伤,因此本文拟就此作用的相关机制进行综述。     

缺血预处理抗缺血再灌注心肌间质损伤的实验研究

目的：观察缺血预处理（ＩＰＣ） 对大鼠缺血再灌注心肌间质胶原及心肌功能结构关系的影响,以进一步探讨ＩＰＣ 对心肌的保护作用。方法：实验采用体重（２５０ ±３０）ｇ 雄性ＳＤ 大鼠２４ 只,分３ 组,每组８ 只,即假手术对照（ＳＣ）组;缺血再灌注（Ｉ／Ｒ） 组和缺血预处理（ＩＰＣ） 组。用超微结构立体计量及羟脯氨酸浓度测定观察心肌间质胶原变化,多道记录仪测量心功能指标,透射电镜观察心肌超微结构。结果：Ｉ／Ｒ 时心肌间质胶原浓度显著低于ＳＣ 组（ Ｐ＜ ０－０１） ,心肌超微结构损伤严重,左室功能明显低于ＳＣ 组（ Ｐ＜ ０－０１） 。ＩＰＣ 组心肌超微结构破坏明显低于Ｉ／ Ｒ 组。同时,心肌间质胶原浓度、胶原纤维线密度及左室功能指标明显高于Ｉ／Ｒ 组（ Ｐ＜ ０－０５ ,Ｐ＜ ０－０１） 。结论：ＩＰＣ 的心肌保护作用不仅表现在对心肌细胞上,而且对心肌间质也有重要的保护作用。     

间充质干细胞衍生的细胞外囊泡在调控心肌缺血再灌注损伤中的研究进展

<正>缺血性心肌病是我国致死率最高的疾病,且随着人民生活水平的改善其的发病率仍在不断攀升^[1]。以经皮冠状动脉介入治疗(percutaneous coro‐nary intervention,PCI)和冠状动脉旁路移植术为代表的血运重建策略是挽救缺血性心肌病患者生命、改善患者远期生存质量的有效手段,特别是随着技术的进步和设备的革新,PCI手术的适用范围不断扩大,已经成为最主要的血运重建手段,     

皮瓣缺血再灌注损伤过程髓过氧化酶及白细胞膜CD18的变化

目的 观察大鼠岛状皮瓣在缺血再灌注损伤过程中髓过氧化酶（MPO）含量及外周血白细胞膜CD18的变化。方法 应用大鼠腹部岛状皮瓣缺血再灌注损伤模型,检测皮瓣髓过氧化酶（MPO）含量,测定外周血白细胞膜CD18水平的变化并作白细胞计数。结果 缺血8h及缺血再灌注1h白细胞膜CD18水平与皮瓣MPO水平较对照组明显增高（P＜0．01）,而缺血再灌注1h较缺血8h组又明显增高（P＜0．01）。缺血再灌注1h组外周血白细胞计数较对照组明显减少（P＜0．01）。结论 大鼠岛状皮瓣缺血再灌注损伤过程中外周血白细胞膜CD18水平增高,白细胞计数减少,皮瓣MPO水平增高。CD18介导的白细胞粘附在皮瓣缺血再灌注损伤中起重要作用。     

缺血预处理对大鼠肥大心脏缺血再灌注损伤的影响

本实验用一氧化氮合成抑制剂－左旋硝基精氨酸复制大鼠高血压心脏肥大模型并作缺血再灌注处理，以观察缺血预处理对高血压肥大心脏Ｉ／Ｒ损伤的影响及ＮＯ在其中的作用。结果显示：ＩＰＣ能减轻大鼠高血压有肥大心脏Ｉ／Ｒ损伤，其程度与ＩＰＣ对正常心脏Ｉ／Ｒ损伤的保护相近，表明高血压肥大心脏同样具有ＩＰＣ保护现象，但ＮＯ在本模型中未起作用。