利多卡因抑制内皮细胞粘附因子表达进而抑制肝癌细胞粘附脐带静脉内皮细胞

【摘要】 目的 探讨利多卡因对血管内皮细胞粘附因子表达的影响。方法 采用不同浓度利多卡因预处理脐带静脉内皮细胞（HUVECs）30 min后,加入肿瘤坏死因子α（TNFα）进行刺激。通过实时荧光定量聚合酶链反应检测选择素E（CD62E）、血管细胞粘附分子-1（VCAM-1）和细胞间粘附分子-1（ICAM-1）表达量,蛋白免疫印迹分析NF-Kappa B（NF-κB）通路蛋白的改变,并通过细胞粘附实验评估利多卡因预处理对肝癌细胞（HepG2）粘附于HUVECs的影响。结果 利多卡因预处理可以明显抑制p65并抑制HepG2粘附于HUVECs。qRT-PCR结果表明利多卡因预处理可明显抑制TNF-α刺激后的CD62E、VCAM-1和ICAM-1表达水平的增高。结论 利多卡因可能通过抑制NF-κB通路进而抑制细胞粘附因子表达并抑制结肝癌细胞粘附于血管内皮细胞。     

Reduction of natural killer and natural killer T cells is not protective in cisplatin-induced acute renal failure in mice

Aims: A recent report showed that fractalkine (CX3CL1), which functions as both a potent chemoattractant and adhesion molecule for monocytes and natural killer (NK) cells was significantly increased in cisplatin‐induced acute renal failure (CisARF) in mice. Therefore, we developed the hypothesis that increased CX3CL1 expression in CisARF initiates NK cell infiltration in the kidney. The aim of the present study was to determine the role of NK cells in CisARF in mice. Methods: Time course of pan‐NK positive cells in CisARF was investigated by using immunohistochemistry (IHC) for CD49b. Pan‐NK positive cells were reduced by using anti‐NK1.1 mAb. The model of pan‐NK positive cells reduction was confirmed by flow cytometry of the spleen and IHC of the kidney. The expression of granzyme A and caspase‐1 was examined, and the activity of caspase‐1 was also determined. We performed a study on whether there was significant protection of renal function after reduction of pan‐NK positive cells. Results: (i) Infiltration of pan‐NK positive cells was prominent on day 3 after cisplatin administration. (ii) granzyme A expression was significantly increased in CisARF and CisARF+NK1.1 Ab compared to vehicle. (iii) Caspase‐1 expression and activity was significantly increased in CisARF mice compared to vehicle and CisARF+NK1.1 Ab. (iv) Reduction of pan‐NK positive cells was not protective in cisplatin‐induced acute renal failure in mice. Conclusions: Although infiltration of pan‐NK cells was significantly increased in CisARF, reduction of infiltration of pan‐NK cells into the kidney was not protective against CisARF in mice.     

二甲基乙二酰基甘氨酸对缺氧复氧诱导的肾小管上皮细胞损伤的保护作用及其机制

目的 通过脯氨酸羟化酶抑制剂二甲基乙二酰基甘氨酸（DMOG）稳定缺氧诱导因子1α（HIF-1α）表达,探讨其对缺氧复氧诱导的肾小管上皮细胞（HKC）损伤的保护作用及其机制。 方法 制作无糖缺氧复氧细胞损伤模型,用不同浓度的DMOG预处理,锥虫蓝染色和乳酸脱氢酶（LDH）活性方法检测细胞活力及损伤;Annexin V和PI染色流式细胞仪技术检测细胞凋亡;实时荧光定量PCR方法检测红细胞生成素（EPO）、热休克蛋白70（HSP70）和血红素氧合酶1（HO-1） mRNA的表达;Western印迹法检测HIF-1α、活性caspase-3和Bcl-2蛋白表达。 结果 正常情况下HKC细胞内几乎无HIF-1α蛋白表达,DMOG刺激6 h后HIF-1α蛋白及其靶基因EPO、HSP70和HO-1 mRNA表达均显著上调（均P ＜ 0.01）,且呈浓度依赖性。500 μmol/L或1 mmol/L DMOG预处理可明显改善缺氧复氧诱导的细胞损伤,表现为细胞存活率升高（95.6%±1.8%、96.1%±1.0%比 83.3%±3.1%）;培养上清液中LDH 活性下降;细胞凋亡减少（8.6%±2.7%、6.1%±2.3%比19.2%±4.0%）（均P ＜ 0.05）。另外,细胞内活性caspase-3蛋白表达显著下调,而Bcl-2蛋白表达则显著上调（均P ＜ 0.05）。 结论 DMOG预处理可稳定肾小管上皮细胞内HIF-1α表达,对缺氧复氧诱导的肾小管上皮细胞损伤具有一定保护作用。其机制可能与促进EPO、HSP70和HO-1表达,抑制caspase-3活化,上调Bcl-2表达有关。     

Expression of cell adhesion molecules in an established and characterized new human renal cell cancer line,CCF-RC7

In order to investigate the importance of cell adhesion molecules (CAMs) in renal cell carcinoma (RCC), a cell line, designated as CCF-RC7, was established from a human RCC of the clear cell type. CCF-RC7 was passaged over 50 times in vitro for 31/2 years. The cell line has an epithelial morphology and a doubling time of 30 h, forming colonies in soft agar with an average efficiency of 10.4% and producing clear cell tumors in athymic nude mice. CCF-RC7 cells have an aneuploid-hypotetraploid karyotype with a modal chromosome number of 82 and rearrangements in chromosomes 9, 12 and 14. Immunohistochemical and flow immunocytometric analyses revealed high expression of ICAM-1 (CD54), and Hermes antigen (CD44), which was significantly upregulated by cytokine and PMA treatment. VLA-4 was expressed on approximately 20% of tumor cells and could not be altered by cytokine or PMA stimulation. High expression of sialyl Lewis X was also demonstrated by immunohistological examination. This newly characterized cell line will serve as a useful model for the study of CAMs during hematogenous metastasis and host defense mechanisms in human RCC.     

胚胎牌细胞不同移植途径对小鼠接种性肉瘤生长和自然杀伤细胞活性的影响

分别于肌肉、腹腔、周围静脉、门静脉对荷瘤小鼠移植胚胎牌细胞，并于移植后30天、60天分别测定瘤体积抑制率和自然杀伤细胞（NK细胞）活性。结果显示：上述4种移植途径在移植后30天均能明显抑制移植性肿瘤生长，NK细胞活性显著升高（P＜0．01）。但移植后60天，仅门静脉组对肿瘤生长保持良好的抑制状态，NK细胞活性保持较高水平，证明经门静脉移植是牌细胞移植最好的途径。     

Antibodies to human adhesion molecules and von Willebrand factor: In vitro cross-species reactivity in the xenotransplantation setting

Abstract: Endothelial cell activation is thought to play an important role in xenograft rejection through cell retraction and expression of pro-coagulant and pro-inflammatory factors. Identification of antibodies recognizing porcine endothelial molecules would be useful to study and manipulate the inflammatory response to a xenograft. The aim of this study was to investigate the cross-reactivity of antibodies directed against human adhesion molecules and von Willebrand factor (vWF). Binding of monoclonal antibodies (mAbs) directed against human CD31, CD44, CD49, CD54, CD62E, CD102, and CD106 was evaluated on resting and activated endothelial cells from human and pig by flow cytometry. Among 30 antibodies tested, 4 were shown to react with pig cells. Two of them, directed against human CD62E (E-selectin) and rabbit CD 106 (VCAM-1) reacted strongly with activated and/or resting pig cells, whereas two others, directed to human CD31 (PECAM) and CD44 (H-CAM), bound weakly to pig cells. In addition, we analyzed the cross-reactivity of five polyclonal or monoclonal antibodies to human or pig vWF with human, baboon, rhesus, pig, and rat vWF. Binding of antibodies was tested by ELISA by using platelet lysates as source of vWF from the different species. Four anti-human or porcine vWF antibodies exhibited a broad reactivity with vWF from all species, whereas one anti-human vWF antibody was specific for primate vWF. In this study, we identified a small number of cross-reacting antibodies that may prove useful to study in vitro and in vivo xenogeneic responses. However, the weak antibody cross-reactivity observed with most porcine molecules points out the necessity of producing species-specific antibodies to study the immune response to xenografts or for use as specific immunosuppressive therapeutic reagents.     

Effect of Shenfu Injection （ ginesenoside and aconite alkaloid） on the apoptosis of intestinal mucosal epithelial cells and its mechanism during ischemia-reperfusion in rats

hegutisnotonlyatargetorganfortraumaandshockbutalsoasatriggerforsystemicinflammationresponsesyndrome .Intestinalmucosalischemiaandreperfusioninjuryisanimportantandessentialetiologicalbasisleadingtomultipleorgandisfunctionsyndrome (MODS) .1Howtopreventandtreatshock ,trauma ,aswellastheischemiaandreperfusioninjuryofthegastrointestinaltracthasbecomethecruxtostudy .Ithasbeenprovedthatcellapoptosisisthemainpatternforthedeathofintestinalepithelialcellsduringischemiaandreperfusion .2 Ithasbeenincrea…     

HLA-Cw4 expression on porcine endothelial cells reduces cytotoxicity and adhesion mediated by CD158a+ human NK cells

Abstract: Background:  Human natural killer (NK) cell-mediated cytotoxicity represents a hurdle in pig-to-human xenotransplantation. It was previously reported that the expression of human major histocompatibility complex class I molecules, including HLA-B27, -Cw3, -E, and -G, partially protects porcine endothelial cells (pEC) from human NK-mediated cytotoxicity and that HLA-G inhibits NK adhesion to pEC. Here, we tested if HLA-Cw4 expression on pEC alone, or concurrently with HLA-Cw3, prevents human NK adhesion and cytotoxicity against pEC via recognition of the killer-cell immunoglobulin-like receptors (KIR) CD158a (KIR2DL1) and CD158b (KIR2DL2/3), respectively.
Methods:  Two pEC lines (2A2 and PEDSV.15) were transfected with HLA-Cw3 and HLA-Cw4. HLA and KIR expression on porcine and human cells were analyzed by flow cytometry. The effect of HLA expression on pEC on human NK-mediated cytotoxicity and adhesion was tested by ⁵¹Cr-release and dynamic adhesion assays, respectively.
Results:  HLA-Cw4 expression on pEC reduced cytotoxicity mediated by CD158a⁺ polyclonal human NK cells by an average of 58%, and by CD158a^bright NK cell clones by 68%, but not by NK cells expressing low levels of CD158. Co-expression of HLA-Cw3 and HLA-Cw4 on pEC did not mediate further protection against NK cytotoxicity. The expression of HLA-Cw4 reduced the adhesion of human NK cells on pEC by a mean of 53%.
Conclusions:  While transgenic expression of HLA-Cw4 on pEC reduces NK cell adhesion and cytotoxicity, co-expression with HLA-Cw3 is not sufficient to completely overcome human NK-mediated cytotoxicity in vitro.     

Clearance of complement by human vascular endothelial cells: effects of hypoxia/reoxygenation and IL-1β activation

Antibody-mediated rejection is characterized by deposits of complement (C) C4 and C3 split products on endothelial cells (ECs). C3 split products are critical mechanistically and diagnostically because they are deposited in amplified quantities, bind covalently to ECs and act as ligands for leukocytes. This study was designed to determine whether cultured vascular human ECs could clear covalently bound C3 split products from their surface. An immunoglobulin M (IgM) antibody against beta(2)-microglobulin of major histocompatibility complex class I antigens was used to activate C in human serum. Some cells were exposed to hypoxia/reoxygenation and/or interleukin 1beta (IL-1beta) prior to incubation with antibody. C3b/iC3b and C3d deposition on the cell surface was measured by flow cytometry. Incubation with antibody followed by human serum caused a dose-dependent deposition of C3b/iC3b and C3d. Over half of deposited C3b/iC3b and one-third of C3d were cleared from the cell surface during a 3-7-h incubation period with human serum. Neither hypoxia/reoxygenation nor IL-1beta further increased the deposition of C3b/iC3b and C3d, and only slightly modulated their rates of clearance. In summary, human ECs rapidly clear iC3b and C3d from their surface. This finding may have important diagnostic and mechanistic implications to transplantation because C3d is used as a marker of antibody-mediated rejection.     

术后自控镇痛不同给药途径对食管癌患者T淋巴细胞亚群及NK细胞的影响

目的观察术后自控镇痛不同给药途径对食管癌患者T淋巴细胞亚群及自然杀伤（NK）细胞的影响。方法开胸食管癌根治手术患者60例,年龄50～65岁,随机分为皮下自控镇痛组（PCSA组）、PCIA组和PCEA组,每组20例。比较三组患者术后6、12、18、24及48h的VAS评分,并在麻醉前（基础值,T1）、术毕（T2）、术后第1天（T3）、第2天（T4）、第5天（T5）采集外周静脉血2ml,采用流式细胞仪检测T淋巴细胞亚群（CD3＋、CD4＋、CD8＋、CD45＋）及NK细胞的水平。结果三组患者VAS评分〈3分。与T1时比较,T3～T5时三组患者CD3＋、CD4＋、CD8＋、NK细胞均下降（P〈0.05或P〈0.01）;T4时PCEA组患者CD3＋、CD4＋、CD8＋水平呈上升趋势,而PCIA组和PCSA组仍呈下降趋势;T5时三组患者CD3＋、CD4＋、CD8＋水平上升恢复,而PCIA组和PCSA组上升更为明显。CD4＋/CD8＋比值从T3时就开始上升恢复,T3时PCEA组达到最大值后开始下降,但较T1时明显上升（P〈0.05）,PCIA组和PCSA组CD4＋/CD8＋比值T3时呈上升趋势,T5时达到最大值,T4、T5时PCSA组CD4＋/CD8＋比值较T1时显著上升（P〈0.05）。T4时PCIA组CD4＋和CD8＋水平较PCEA组降低（P〈0.05）。结论 PCSA、PCIA和PCEA都适用于食管癌患者的术后镇痛,PCEA免疫功能恢复较PCIA早,PCSA与PCEA对术后免疫功能的影响无明显不同。     

Soluble saccharides block the inhibition of agonist-induced human platelet aggregation observed after in vitro incubation of human platelet-rich plasma with porcine aortic endothelial cells

Platelet aggregation is a prominent feature in the hyperacute process of vascularized allografts and xenografts. In a study of extracorporeal connection of pig kidneys to the blood circulation of human volunteers, we observed in one case considerable destruction of human platelets in the pig kidney without signs of hyperacute rejection or microthrombi formation. In the present study, we have investigated the agonist-induced aggregation of human platelets in mixtures with porcine aortic endothelial cells (PAEC). In vitro incubation of human platelet-rich plasma (PRP) with PAEC inhibited platelet aggregation induced by ADP, collagen and arachidonic acid in a time-dependent manner and partially inhibited adrenalin-induced aggregation. Aggregation of the human platelets could not be induced by high concentrations of ADP (20 μM) to overcome the inhibition capacity of the PAEC. The PAEC inhibiting effect could be transferred by the supernatants of PAEC/PRP and PAEC/PPP incubation mixtures. Preincubation of the PAEC with aspirin, but not with N^G-methyl-L-Arg, reduced the aggregation inhibitory effect. Control experiments mixing human umbilical vein endothelial cells (HUVEC) and human PRP or mixing porcine PRP and PAEC did not elicit any inhibition of ADP-induced platelet aggregation. The aggregation inhibition effect could partially be blocked by preincubation of PRP with soluble Galα1–3Gal, Galα1–3β1–4GlcNAc, lactose, galactose, and glucose, but not by lactosamine, galactosamine, or glucosamine. The Galα1–3Gal disaccharide was most effective in blocking aggregation inhibition, and to a similar extent as its ability to block the human anti-pig lymphocytotoxicity reaction. In conclusion, the data indicate that PAEC, upon stimulation by human anti-pig xenoantibodies in a nondynamic system, inhibits agonist-induced human platelet aggregation, and that this effect is probably at least partially caused by prostacyclin released from the PAEC. Received: 12 August 1997 Received after revision: 19 March 1998 Accepted: 15 April 1998     

热打击及中暑小鼠血清对小鼠肺微血管内皮细胞黏附单核细胞能力的影响

目的：研究热打击及10%中暑小鼠血清刺激肺微血管内皮细胞(PMVECs)对 PMVECs 表达血管内皮黏附分子(VCAM-1)和细胞间黏附分子(ICAM-1)水平及其黏附单核细胞能力的影响。方法：根据二次磁珠分选法分离乳鼠PMVECs,光镜下观察细胞形态并对其血管内皮细胞钙黏蛋白(VE-Cadherin)及Ⅷ因子进行染色鉴定。将PMVECs随机分为3组,分别进行37℃正常培养(对照组)、给予42℃2 h热打击或10%中暑小鼠血清刺激,24 h后分别提取总 RNA,荧光定量PCR及 Weston blot检测 VCAM-1及 ICAM-1mRNA和蛋白的表达水平,荧光显微镜观察其对人单核细胞(THP-1细胞)的黏附能力。结果：所分离 PMVECs 纯度较高,光镜下细胞呈特征性鹅卵石形状排列,VE-Cadherin及Ⅷ因子均可染色。与对照组相比,给予42℃2 h的热打击24 h后,VCAM-1及 ICAM-1的 mRNA 及蛋白水平表达降低(P<0.05),而给予10%中暑小鼠血清刺激24 h 后VCAM-1及ICAM-1的mRNA及蛋白水平表达增加(P<0.01)。与对照组相比,热打击组黏附THP-1细胞的细胞数无明显变化,中暑小鼠血清刺激组THP-1细胞数明显增多,显示PMVECs对单核细胞的黏附能力增强。结论：单独热打击并未使小鼠PMVECs黏附分子表达增加,而给予中暑小鼠血清刺激后其黏附分子表达显著上调,从而使单核细胞易于附壁,导致或加重了中暑相关的急性肺损伤。     

丙泊酚静脉麻醉对行取卵术患者自然杀伤细胞数量及妊娠结局的影响

目的:评价丙泊酚静脉麻醉对行取卵术患者自然杀伤（natural killer, NK）细胞的数量及妊娠结局的影响。方法:选择择期接受体外受精-胚胎移植（in vitro fertilization and embryo transfer, IVF-ET）的患者110例,年龄20~40岁,ASA分级Ⅰ、Ⅱ级,采用随机数字...     

The effects of molsidomine on hypoxia inducible factor alpha and Sonic hedgehog in testicular ischemia/reperfusion injury in rats

This study was designed to determine the effect of molsidomine (MO), a precursor of nitric oxide (NO) donor, on hypoxia inducible factor alpha (HIF-1α) and Sonic hedgehog (Shh) levels considered to be involved in the development of testes ischemia/reperfusion (I-R) injury. Torsions were created by rotating ipsilateral testes 720° in a clockwise direction for 6 h and 1-h detorsion of the testis was performed. A sham operation was performed in group 1 (control, n = 7). In group 2 (I-R/Untreated, n = 7), following 6 h of unilateral testicular torsion, 1-h detorsion of the testis was performed. No drug was given. In group 3 (I-R/MO), after performing the same surgical procedure as in group 2, a NO donor MO was given at the starting time of reperfusion. In group 4 (I-R/L-NAME), after performing the same surgical procedure as in group 2, L-NAME was given at the starting time of reperfusion. Testes malondialdehyde (MDA) levels were determined as well as examining the testes histologically. Treatment of rats with MO produced a significant reduction in the levels of MDA and histopathological score compared to testes I-R groups. The Sonic hedgehog (Shh) expression in the basement membrane of the tubuli seminiferi, and sertoli and germinal cells in testicular tissue, were greatly increased in the I-R/MO group compared to groups 1, 2 and 4. Additionally, the HIF-1α expression in the interstitial spaces in testicular tissue were greatly increased in the I-R/MO group. The results suggest that MO has a protective effect against ischemia/reperfusion injury in rat testes and may affect Shh and HIF-1α signaling pathway.     

肿瘤坏死因子α对人腹膜间皮细胞基质金属蛋白酶及其组织抑制物表达的影响

目的 观察肿瘤坏死因子α（TNF-α）对人腹膜间皮细胞(HPMC) 基质金属蛋白酶（MMP）2、MMP-9及其组织抑制物（TIMP）2、TIMP-1 mRNA和Ⅰ型胶原表达的影响，同时观察TNF-α、TGF-β、IL-1单独或协同作用对HPMC的MMP-9活性的影响。 方法 采用半定量RT-PCR法测定细胞MMP-2、MMP-9、TIMP-2及TIMP-1 mRNA 的表达。采用Biotrak MMP-9 活性检测系统来精确定量测定MMP-9活性及MMP-9原的含量。ELISA法检测Ⅰ型胶原蛋白的表达。 结果 TNF-α(1~10 μg/L)分别刺激HPMC 4、16、24及48 h后， HPMC的MMP-9 mRNA表达显著上调，为基础的2.3～4.9倍（P ＜ 0.05），呈时间依赖性。TNF-α（1 μg/L）作用48 h后显著下调TIMP-1、TIMP-2 mRNA 表达，为基础的77.2％、61.3％（P ＜ 0.05），而MMP-2 mRNA表达没有显著变化。TNF-α+TGF-β１ （1~10 μg/L）、 TNF-α+TGF-β1+IL-1(1~10 μg/L)和TNF-α(5~10 μg/L)刺激HPMC 24 h后，促其分泌MMP-9 的作用最为明显。同时，TNF-α明显上调Ⅰ型胶原蛋白表达（P ＜ 0.05）。 结论 TNF-α 明显上调HPMC的MMP-9 mRNA的表达和MMP-9 的活性；联合其他细胞因子后促MMP-9分泌的作用更明显。TNF-α单独或协同其他因子在腹膜纤维化的过程中可能发挥了重要作用。     

血小板反应蛋白-1在PCOS患者中对卵巢血流调控作用及其对人卵巢微血管内皮细胞增殖能力的影响

目的 探讨PCOS患者血清中血小板反应蛋白-1(TSP-1)表达水平与卵巢间质血供之间的关系,以及TSP-1对体外培养的人卵巢微血管内皮细胞(HOMEC)增殖能力的影响.方法 选取2018年10月至2019年10月在我院生殖中心就诊的女性,50例诊断为PCOS的患者纳入PCOS组,50例因男方因素不孕的健康女性纳入对照...     

人胎盘MSCs移植对急性肺损伤小鼠肺血管内皮通透性及肺组织损伤修复的影响

目的探讨人胎盘MSCs(human placental MSCs,hPMSCs)移植对脂多糖诱导的急性肺损伤(acute lung injury,ALI)小鼠肺血管内皮通透性及肺组织损伤修复的影响。方法取人胎盘组织采用酶消化法分离培养hPMSCs并传至第3代,流式细胞仪检测细胞表型。将24只6周龄雄性C57BL/6小鼠随机分成3组(n=8)。其中,ALI模型组及h PMSCs治疗组采用气道滴注脂多糖方法制备ALI模型,对照组滴注生理盐水;滴注脂多糖12 h后hPMSCs治疗组尾静脉注射第3代hPMSCs,ALI模型组与对照组注射生理盐水。注射24 h后取各组小鼠肺组织,HE染色观察肺组织病理改变,测量肺组织湿/干质量比(wet/dry mass ratio,W/D),伊文思蓝渗漏实验检测小鼠肺血管内皮通透性,Western blot检测肺组织通透性相关蛋白血管内皮钙黏蛋白(VE-cadherin)的表达。结果流式细胞仪检测显示,分离培养的细胞具有典型MSCs表型特征。各组小鼠均存活。ALI模型组肺泡结构明显塌陷、大量炎性细胞浸润、局部肺泡出血;hPMSCs治疗组肺泡塌陷明显改善,炎性细胞浸润明显减少,肺间质有少许红细胞。ALI模型组肺组织W/D、伊文思蓝染液含量明显高于对照组及hPMSCs治疗组,hPMSCs治疗组高于对照组,差异均有统计学意义(P<0.05)。ALI模型组VE-cadherin蛋白相对表达量明显低于对照组及hPMSCs治疗组,hPMSCs治疗组低于对照组,差异均有统计学意义(P<0.05)。结论静脉注射hPMSCs可有效降低脂多糖介导的肺血管内皮通透性增加,减轻肺组织损伤程度,对小鼠ALI具有治疗作用。     

BMSCs移植对大鼠脊髓损伤后VEGF基因和血管生成的影响

目的研究BMSCs移植对大鼠脊髓损伤(spinal cord injury,SCI)后运动功能恢复、VEGF基因表达和血管生成的影响,探讨BMSCs治疗SCI的机制。方法取5只4周龄Wistar雄性大鼠骨髓,分离培养BMSCs,取第3～4代细胞进行实验。取Wistar雌性成年大鼠87只,体重220～250 g,随机分为假手术组(A组,21只)、DMEM注射组(B组,33只)和BMSCs移植组(C组,33只)。A组仅咬除T8～10棘突和椎板;B、C组参照Nystrom方法制备T8～10 SCI模型,伤后30 min C组注射BMSCs(共3.0×105个),B组注射等量DMEM。于术后3、7、14、28 d采用Basso-Beattie-Bresnahan(BBB)评分法评价大鼠后肢运动功能;术后3、7、14、28 d应用Ⅷ因子免疫组织化学染色行微血管计数观测血管生成情况;术后1、3、5 d应用RT-PCR法检测损伤脊髓组织内VEGF mRNA的表达。结果术后各时间点B、C组大鼠后肢功能随时间延长均有不同程度恢复,各时间点BBB评分与A组比较差异均有统计学意义(P<0.05)。术后28 d,C组BBB评分显著高于B组(P<0.05),其余时间点B、C组比较差异无统计学意义(P>0.05)。术后3 d,B、C组损伤周围区脊髓灰质腹侧角内微血管数目明显少于A组(P<0.05);术后7、14、28 d与A组比较差异无统计学意义(P>0.05)。C组术后3、7 d脊髓灰质腹侧角内微血管数目与B组比较差异无统计学意义(P>0.05);但术后14、28 d显著高于B组(P<0.05)。各组术后各时间点损伤周围区脊髓白质内的微血管数目比较差异均无统计学意义(P>0.05)。RT-PCR检测示,A组大鼠脊髓组织内VEGF mRNA表达水平较低。B、C组与A组相比,于术后1 d开始VEGF mRNA表达增加,3 d时表达达峰值,5 d时表达下降但仍高于A组,各时间点与A组比较差异均有统计学意义(P<0.05);C组各时间点均高于B组(P<0.05)。结论 BMSCs可能通过上调VEGF基因表达和增加脊髓内新生血管而促进大鼠SCI后运动功能恢复。