不同方式移植骨髓间充质 干细胞治疗急性重症胰腺炎大鼠的疗效对比

目的探索骨髓间充质干细胞(bMSCs)治疗大鼠急性重症胰腺炎(SAP)模型的高效移植方式。方法将大鼠随机分为对照组(control)、急性重症胰腺炎组(SAP)(胰胆管内逆行注射5%硫磺胆酸钠)。SAP模型制作成功6 h后,分为尾静脉骨髓间充质干细胞移植组(tail vein)、腹腔注射移植组(intraperitoneal)和尾静脉-腹腔注射联合移植组(tail vein+intraperitoneal),每组18只。在移植后第24、48和72 h分别安乐死6只大鼠,收集胰腺组织及血清,使用ELISA试剂盒测定血清中抗炎因子IL-10及炎性因子IL-1β、TNF-α和IL-6的水平;HE染色后光学显微镜下观察胰腺组织病理和第72 h后各组大鼠小肠黏膜及肺泡组织病理改变;冰冻切片在荧光显微镜下观察DAPI标记的bMSCs在胰腺组织的分布。结果 SAP组胰腺组织有大量出血、水肿、炎性反应及坏死;与对照组相比, 3种不同方式bMSCs移植组胰腺组织的出血、水肿、炎性反应及坏死明显减少(P0.05),IL-1β、TNF-α和IL-6水平显著降低,IL-10水平显著升高(P0.05);尾静脉+腹腔联合移植组较单纯尾静脉或腹腔移植组胰腺组织的出血、水肿、炎性反应及坏死进一步减少(P0.05),IL-1β、TNF-α和IL-6水平进一步降低(P0.05),IL-10的水平进一步升高(P0.05);DAPI标记的bMSCs在移植组的胰腺组织内均有分布,且在尾静脉+腹腔联合移植组观察到更多的荧光分布;72 h后可见移植组小肠黏膜及肺泡组织完整性较SAP组明显好转,中性粒细胞浸润及出血情况较SAP组明显减轻,相比于腹腔和单纯尾静脉移植,联合移植组情况进一步改善。结论在大鼠SAP模型中,bMSCs通过尾静脉+腹腔联合移植能显著抑制炎性反应,减少SAP相关性胰腺、小肠黏膜及肺损伤,其效果显著优于单纯尾静脉移植和腹腔注射移植。     

间充质干细胞治疗急性肺损伤的研究进展

间充质干细胞是一种多能造血干细胞。在急性肺损伤中,它可以分化为Ⅰ和Ⅱ型上皮细胞,修复受损的局部组织。此外,间充质干细胞还通过减少促炎症细胞因子IL-1、MIP-2、INF-γ和TNF-α的释放,上调抗炎症细胞因子IL-10、IL-1ra和IL-13的分泌,减轻炎症细胞反应,从而发挥治疗作用。     

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.     

间充质干细胞对固有免疫细胞调节作用的研究进展

近年来研究发现,间充质干细胞除具有支持体外造血、促进体内造血功能重建外,还具有较低的免疫原性,能够通过调节多种固有或适应性免疫细胞的功能,来调节机体的免疫反应。本文阐述了间充质干细胞对巨噬细胞、中性粒细胞、自然杀伤细胞以及树突状细胞的免疫调节作用。     

骨髓间充质干细胞异体移植免疫反应的研究进展

骨髓间充质干细胞（BMSC）具有独特的生物学特性，其免疫抑制作用引起了人们广泛的兴趣。大量资料表明，MSC能在同种异基因，甚至异种基因的环境中长期存活，并且保持它的多向分化潜能。这一特性使MSC异体移植预防和治疗免疫性疾病成为可能。因此有必要从问充质干细胞的低免疫原性、灯免疫细胞及补体系统的调节和诱导抑制性免疫微环境等方面探讨BMSC避免同种异体移植排斥反应的可能机制。     

间充质干细胞与自身免疫病研究进展

间充质干细胞(MSCs)是干细胞家族的重要成员,其源于发育早期的中胚层和外胚层.MSCs最初在骨髓中发现,因其具有多向分化潜能、造血支持和自我复制等特点而日益受到人们的关注.近年来,因其低免疫原性和免疫调节作用的研究发现,进而揭开了MSCs在治疗自身免疫性疾病中的广阔前景.因而研究MSCs、表面标志、诱导分化、免疫调控作用以及在自身免疫性疾病中的应用和机制具有重要意义.     

骨髓间充质干细胞免疫学特性的研究进展

随着免疫学和移植学的发展,人类对自身免疫性疾病认识的不断提高和对器官移植的日益接受,免疫抑制剂在临床医学中的应用愈来愈广泛。它们能提高患者的生存质量,延长患者的寿命,但是其昂贵的费用、易引起感染和肿瘤等的副作用明显地限制了它的临床应用。为此,免疫学专家和移植学专家一直在探索理想的诱导特异性耐受的方法和途径,以便减少免疫抑制剂的应用剂量和副作用,乃至部分或甚至完全替代免疫抑制剂。近20多年来,成体干细胞的可塑性现象引起人们的极大兴趣。其中骨髓间充质干细胞(bone marrow mesenchymal stemcell,MMSC)的功能作用和…     

间充质干细胞作为免疫调节剂用于临床肝移植治疗研究进展

间充质干细胞(mesenchymal stem cell,MSC)因其多潜能性和可扩增性对再生医学产生了不可估量的影响.除可塑性外,MSC作为新型免疫调节治疗产物正在被应用于临床前及临床研究中.无论是体外试验还是疾病模型中MSC均显示了良好的免疫抑制功能,为正在遭受自身免疫病和移植排斥反应困扰的人们带来了福音.     

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。     

间充质干细胞免疫调节特性与临床应用的研究进展

间克质于细胞(mesenchymal stem cell,MSC)是可以从骨髓、脂肪、脐带、骨膜等组织分离而来的一种干细胞,具有黏附培养基生长的特性和复杂的免疫表型,可以向3个胚层细胞分化.MSC在组织修复和再生方面有良好的治疗效果.MSC回输体内可以定位到受损的组织,并且通过上调抗炎症细胞因子和下调促炎症细胞因子,调节受损部位的炎症反应.此外,MSC有显著的免疫调节特性,可以抑制T细胞、NK细胞等细胞的功能,调节树突状细胞的活性而诱导免疫耐受,在临床上具有广泛的应用前景.     

Therapeutic effect of human umbilical cord-derived mesenchymal stem cells in rat severe acute pancreatitis

Aim: To investigate the therapeutic effect of umbilical cord-derived mesenchymal stem cells (UC-MSCs) on rat severe acute pancreatitis (SAP). Methods: Rats were randomly divided into three groups (n = 15 per group): control group, SAP group, and SAP+MSCs group. SAP was established by retrograde pancreatic duct injection of 3% sodium taurocholate. In SAP+MSCs group, UC-MSCs at 1×10⁷ cells/kg were injected via the tail vein 12 h after SAP. Rats (n = 5 per group) were sacrificed on days 1, 3 and 5, and the blood and pancreatic tissues were collected. The levels of serum amylase, lipase, inflammatory cytokines, and anti-inflammatory cytokines were determined. Pathological changes of the pancreas (HE staining) and apoptotic acinar cells (TUNEL staining) were observed under light microscope. Results: The levels of serum amylase and lipase in SAP group were significantly higher than those in control group (P<0.05). The pancreas in SAP group showed significantly massive edema, inflammation, hemorrhage and necrosis when compared with control group. There were numerous TUNEL-positive apoptotic acinar cells after SAP. However, in SAP+MSCs group, the levels of serum amylase were significantly reduced on days 1, 3, and 5 after MSC transplantation (P<0.01). The serum lipase level in SAP+MSCs group was significantly lower than that in SAP group on days 3 and 5 (P<0.01). The edema formation, inflammatory cell infiltration, hemorrhage, and necrosis were reduced significantly attenuated in SAP+MSCs group as compared to SAP group (P<0.05). MSCs significantly reduced the levels of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), but increased the levels of anti-inflammatory cytokines (IL-4 and IL-10) in SAP rats. The number of TUNEL-positive acinar cells was significantly reduced on days 3 and 5 after MSCs transplantation (P<0.01). Conclusion: Transplantation of UC-MSCs significantly inhibits inflammation and decreases pancreatic injury secondary to SAP.     

Angiopoietin-1 gene-modified human mesenchymal stem cells promote angiogenesis and reduce acute pancreatitis in rats

Mesenchymal stem cells (MSCs) can serve as a vehicle for gene therapy. Angiopoietin-1 (ANGPT1) plays an important role in the regulation of endothelial cell survival, vascular stabilization, and angiogenesis. We hypothesized that ANGPT1 gene-modified MSCs might be a potential therapeutic approach for severe acute pancreatitis (SAP) in rats. Human umbilical cord-derived MSCs with or without transfection with lentiviral vectors containing the ANGPT1 gene were delivered through the tail vein of rats 12 h after induction of SAP. Administration of MSCs alone significantly reduced pancreatic injury and inflammation, as reflected by reductions in pancreatitis severity scores and serum amylase and lipase levels as well as reducing the serum levels of proinflammatory cytokines (TNF-α, IFN-γ, IL-1β, and IL-6). Furthermore, administration of ANGPT1-transfected MSCs resulted in not only further reductions in pancreatic injury and serum levels of proinflammatory cytokines, but also promotion of pancreatic angiogenesis. These results suggest that MSCs and ANGPT1 have a synergistic role in the treatment of SAP. ANGPT1 gene-modified MSCs may be developed as a potential novel therapy strategy for the treatment of SAP.     

Role of bone marrow mesenchymal stem cells in L-arg-induced acute pancreatitis: effects and possible mechanisms

Objectives: Mesenchymal stem cells (MSCs) have shown an obvious protective effect on systemic inflammation. The purpose of this study is to assess the effect and possible mechanism of bone marrow MSCs (bmMSCs) on acute pancreatitis (AP). Methods: BmMSCs of SD rats were isolated and cultured in vitro. L-Arginine-induced acute pancreatitis was used as AP model in vivo. Pancreatic injury was assessed by serum amylase, lipase, cytokines and pancreatic histology. RT-PCR was applied to investigate mRNA expression of pancreas tissue. Western-blot and immunohistochemistry (IHC) were applied to test the role of NF-κB p65 signaling pathway. Tracking and Positioning of CM-Dil labeled bmMSCs in vivo was further studied. Results: Treatment with bmMSCs attenuated acute pancreatic injury and AP-associated lung injury obviously, with decreased serum IL-1β, IL-6, TNF-α, down-regulated expressions of IL-1α, IL-6, TNFα in pancreas tissue and reduced nuclear translocation of NF-κB p65 in AP. Localization of bmMSCs in vivo was due to being passively trapped in related organs, but not actively homing to inflammatory sites of pancreas during the early phase of AP. Conclusions: Taken together, the results showed that bmMSCs played a protective role in AP in many aspects, which might protect against experimental pancreatitis partly by regulating release of inflammatory cytokines by an exocrine secretion.     

同种异体骨髓间充质干细胞移植治疗急性肝功能衰竭

背景：对于急性肝功能衰竭的临床治疗,目前尚缺乏特效手段。骨髓间充质干细胞在特定环境下可分化为具有部分肝功能的类肝样细胞,从而参与肝功能的修复和重构,为急性肝功能衰竭的治疗提供了新的手段。 目的：评价同种异体骨髓间充质干细胞移植治疗大鼠急性肝功能衰竭的疗效,为其临床应用提供理论依据。 方法：采用全骨髓培养法培养并纯化其骨髓间充质干细胞。30只健康SD大鼠随机均分为3组：正常对照组：不予任何处理;肝衰竭组和移植组：用腹腔注射四氯化碳石蜡油溶液的方法复制鼠急性肝功能衰竭模型后24 h分别经其尾静脉注射生理盐水和等量骨髓间充质干细胞。在移植后第1,2,3,7天抽血检测其肝功能,并取肝脏行病理学检查。 结果与结论：急性肝功能衰竭鼠经骨髓间充质干细胞移植治疗后生存率为70%,与肝衰竭组大鼠存活率20%相比差异有显著性意义(P < 0.05);肝功能指标中谷丙转氨酶和谷草转氨酶相比,移植组明显低于肝衰竭组,差异有显著性意义(P < 0.05)。肝脏组织病理学结果显示移植组肝细胞变性及坏死程度以及炎症浸润程度轻于肝衰竭组。因此,经尾静脉移植骨髓间充质干细胞能提高急性肝功能衰竭大鼠的生存率、改善肝功能及减轻肝脏坏死程度,对大鼠急性肝功能衰竭有一定的治疗作用。     

脐带血间充质干细胞的研究进展

脐带血中存在着丰富的造血干细胞,在移植方面发挥重要作用,在脐血中是否还存在间充质干细胞却有争议,有的学者认为其中有间充质干细胞,且与骨髓间充质干细胞(mesenchymalstemcells,MSC)的形态、表面标志及分化潜能非常相似,有的学者认为含量较低,难以传代培养扩增,总结了近5年来国内外关于脐血间充质干细胞的研究情况,为脐血的充分利用提供更多的资料。     

骨髓间充质干细胞对肝病的治疗作用

骨髓间充质干细胞（MSCs）是一群具有多向分化潜能的成体干细胞,具有抑制T细胞活化、调节异体移植的免疫耐受等免疫特性。肝移植是各种晚期肝病的主要治疗方法,但由于器官来源的缺乏和伴随的免疫排斥反应,其应用受到限制。研究证明骨髓是肝祖细胞的来源之一,在一定的条件下,骨髓间充质干细胞能够分化为功能性肝细胞并分泌相关细胞因子,促进肝细胞的再生,为临床治疗各种肝损害提供了新的途径。     

骨髓间充质干细胞治疗大鼠急性心肌梗死

目的:比较经不同途径移植骨髓间充质干细胞(BMSCs)治疗大鼠急性心肌梗死(AMI)的疗效,探求更为适合的移植途径。方法:采集大鼠BMSCs,并进行培养及鉴定。以5-氮胞苷(5-aza)10μmol/L,诱导BMSCs为心肌样细胞并鉴定。建立大鼠急性心肌梗死模型并鉴定。对照组不予处理,静脉移植组和心外膜移植组分别经尾静脉和心外膜移植心肌样细胞悬液200μl(心肌样细胞5×10~6),联合移植组同时经尾静脉和心外膜分别移植心肌样细胞悬液各200μl。4周后观察SD大鼠心肌组织形态及蛋白表达情况。结果:静脉移植组、心外膜移植组、联合移植组的心肌梗死区域均可见有DAPI标记阳性的移植细胞,其中联合移植组较静脉移植组、心外膜移植组数量明显增多;H-E染色可见后三组较对照组梗死区域心肌细胞排列整齐,细胞核较完整,联合移植组梗死改善程度明显;后三组较对照组血管紧张素转换酶2(ACE2)表达水平升高,联合移植组较静脉移植组、心外膜移植组升高水平更显著。结论:经静脉、心外膜和两者联合移植诱导后的BMSCs均能促进梗死部位的组织修复,抑制心室重构,其中联合移植途径治疗效果更明显。     

间充质干细胞治疗骨关节炎的临床研究进展

骨关节炎(OA)是一种进展性疾病,表现为软骨缺损、滑膜炎症、软骨下骨硬化及骨赘形成,关节表面软骨再生能力弱,无法实现自我修复.骨关节炎患者的主要临床表现为疼痛和关节活动受限,肢体功能和生活质量受影响.理想的治疗方案应缓解疼痛、促进软骨修复,以延缓或阻止病程进展.间充质干细胞(MSCs)在治疗骨关节炎方面具备以上优势且无...