不同间歇时间磁刺激对星形胶质细胞迁移能力的影响及机制分析

目的 探讨不同间歇时间磁刺激对星形胶质细胞迁移能力的影响及相关机制。 方法 将传代星形胶质细胞分为对照组、1 s间歇组、5 s间歇组和10 s间歇组,分别给予相应间歇时间磁刺激,观察不同间歇时间磁刺激对星形胶质细胞迁移能力的影响;星形胶质细胞在磁刺激作用下,采用PEA-15磷酸化阻滞剂Bis I、细胞外调节蛋白激酶（ERK1/2）阻滞剂U0126处理细胞,采用Transwell实验检测星形胶质细胞迁移能力,采用Western blot技术检测pPEA-15和pERK1/2表达。 结果 1 s间歇时间磁刺激可明显增强星形胶质细胞迁移能力,促进PEA-15及ERK1/2磷酸化,并提高基质金属蛋白酶-9（MMP-9）表达;加入Bis I可降低ERK1/2磷酸化水平及MMP-9表达,减弱磁刺激对星形胶质细胞的促迁移作用;经U0126试剂处理后,发现磁刺激对星形胶质细胞的促迁移作用显著下降。 结论 1 s间歇时间磁刺激能促进星形胶质细胞PEA-15磷酸化,提高ERK1/2磷酸化水平,进而增强下游蛋白MMP-9表达,从而促进星形胶质细胞迁移。     

HTRA1基因过表达对视网膜色素上皮细胞增殖及迁移影响

目的 研究HTRA1基因过表达对人视网膜色素上皮细胞(RPE)的增殖和迁移能力的影响,探讨HTRA1基因在老年黄斑变性(AMD)发病中的作用.方法 用携带人HTRA1基因的慢病毒感染体外培养的人RPE细胞,荧光显微镜下观察细胞的感染效率,RT-PCR和Westem Blot检测HTRAlmRNA和蛋白的表达,MTT比色法检测感染细胞毒性及细胞增殖能力,TransweH小室检测细胞迁移能力变化.结果 与空载体慢病毒感染RPE细胞相比,HTRA1基因慢病毒感染的RPE细胞HTRA1mRNA和蛋白的表达量明显增加,细胞增殖能力及迁移能力均明显下降,差异有统计学意义(P＜0.01).结论 过表达HTRA1基因能抑制人视网膜色素上皮细胞的增殖及迁移,THRA1基因可能通过影响RPE细胞功能参与AMD的进程.     

The effect of delivering the chemokine SDF-1α in a matrix-bound manner on myogenesis

Several chemokines are important in muscle myogenesis and in the recruitment of muscle precursors during muscle regeneration. Among these, the SDF-1α chemokine (CXCL12) is a potent chemoattractant known to be involved in muscle repair. SDF-1α was loaded in polyelectrolyte multilayer films made of poly(l-lysine) and hyaluronan to be delivered locally to myoblast cells in a matrix-bound manner. The adsorbed amounts of SDF-1α were tuned over a large range from 100 ng/cm² to 5 μg/cm², depending on the initial concentration of SDF-1α in solution, its pH, and on the film crosslinking extent. Matrix-bound SDF-1α induced a striking increase in myoblast spreading, which was revealed when it was delivered from weakly crosslinked films. It also significantly enhanced cell migration in a dose-dependent manner, which again depended on its presentation by the biopolymeric film. The low-crosslinked film was the most efficient in boosting cell migration. Furthermore, matrix-bound SDF-1α also increased the expression of myogenic markers but the fusion index decreased in a dose-dependent manner with the adsorbed amount of SDF-1α. At high adsorbed amounts of SDF-1α, a large number of Troponin T-positive cells had only one nucleus. Overall, this work reveals the importance of the presentation mode of SDF-1α to emphasize its effect on myogenic processes. These films may be further used to provide insight into the role of SDF-1α presented by a biomaterial in physiological or pathological processes.     

Reduced levels of transforming growth factor-beta1, interleukin-12 and increased migration inhibitory factor are associated with severe malaria

In the present study, we investigated plasma levels of interleukin (IL)-12 and transforming growth factor (TGF-beta1) in malaria patients as these two cytokines regulate the balance between pro- and anti-inflammatory cytokines. We compared plasma IL-12 and TGF-beta1 levels in groups of malaria patients categorized as uncomplicated, severe, cerebral and placental malaria. Both TGF-beta1 and IL-12 levels were significantly reduced in peripheral plasma of adults with severe and cerebral malaria as well as in plasma of Tanzanian children with cerebral malaria (P<0.05). Similar results were observed with both placental and peripheral plasma of pregnant women who were infected with Plasmodium falciparum. IL-18, a cytokine known to be critical for the induction of IFN-gamma along with IL-1, was produced more in uncomplicated adult patients than in aparasitimic healthy controls (P<0.05). However, IL-18 response rate declined as the symptoms of the disease became more severe suggesting that the IL-18 response may be impaired with increased malaria severity. Together, the results of the three cytokines support the notion that imbalance between pro- and anti-inflammatory cytokines may contribute to the development of severe malaria infection. With malaria infection during pregnancy, we demonstrated that macrophage migration inhibitory factor (MIF) levels in infected placental plasma were significantly higher than those in the paired peripheral plasma (P<0.05). MIF, therefore, may play an important role in the local immune response to placental P. falciparum infection.     

家兔腹腔巨噬细胞能产生单核细胞趋化蛋白-1

用培养的兔腹有空巨噬细胞条件培养基作为趋化因子的来源，用改良的Ｂｏｙｄｅｎ小室微孔滤膜法进行单核细胞趋化试验，观察了兔腹腔巨噬细胞条件培养基对单核细胞的趋化作用和抗单核细胞趋化蛋白－１抗体对单核细胞迁移的影响，同时用Ｎｏｒｔｈｅｒｎ ｂｌｏｔ分析方法检测了ＭＣＰ－１ ｍＲＮＡ在该巨噬细胞的表达。结果显示，该条件培养基对单核细胞有明显的趋化作用，并被抗ＭＣＰ－１抗体所抑制，同时该巨噬细胞也能表达ＭＣ     

Bmi1对舌鳞癌侧群细胞迁移、侵袭和增殖能力的调控作用及机制探讨

目的: 探讨Bmi1如何调控舌鳞癌侧群细胞的迁移、侵袭和增殖能力。 方法: 首先采用Hoechst33342法,利用流式细胞仪分选舌鳞癌细胞株UM1（高转移株）中的侧群细胞（side population cell, SP）和非侧群细胞（non-side population cell, Non-SP）。Transwell 实验检测沉默Bmi1后SP细胞的迁移、侵袭能力。球囊形成和平板克隆实验检测沉默Bmi1后SP细胞的球囊和克隆形成率;CCK8 实验检测沉默Bmi1后SP细胞的增殖能力。Western免疫印迹检测沉默Bmi1后SP细胞中侵袭、转移相关基因（SOD2、Slug）及干细胞标志物（ABCG2、Nanog）的表达水平。采用SPSS17.0软件包对数据进行统计学处理。 结果: 转染Bmi1 siRNA使SP细胞中Bmi1表达水平下调后,SP细胞的迁移和侵袭能力显著下降;球囊形成率和克隆形成率也显著低于对照组;增殖能力受到抑制;SOD2、Slug和干细胞标志物ABCG2和Nanog的表达水平显著下降。 结论: 沉默Bmi1可调控舌鳞癌侧群细胞的迁移、侵袭和增殖。     

Knocking down the expression of adenylate cyclase-associated protein 1 inhibits the proliferation and migration of breast cancer cells

Adenylate cyclase-associated protein 1 (CAP1) is a conserved protein that was found to be up-regulated in breast cancer and related to the migration of breast cancer. We verified its roles in breast cancer specimens and cell lines. In our results, 71 of 100 specimens of breast cancer showed high levels of CAP1 by immunohistochemistry. Associated with statistical analysis, we saw that CAP1 was related to the grade of breast cancer. In MDA-MB-231, the expression of CAP1 was the highest and by knocking down the expression of CAP1 in MDA-MB-231, its ability for proliferating and migrating apparently decreased and induced changes in morphology, which were related to the arrangement of F-actin. Therefore, CAP1 might be a potential molecular targeted therapy for surgery and immune treatment.     

Regulation of Migration of Human Colonic Myofibroblasts

Background and Aim: The migration of mesenchymal cells to areas of mucosal or submucosal tissue damage is an essential factor for wound healing in the intestine. Thus far, neither migration inducing factors nor signal transduction cascades involved in the migration of colonic myofibroblasts (CMF) have been studied in detail. Methods: Primary CMF were isolated from the mucosa of surgical specimens or endoscopic biopsies. Migration assays of CMF were performed in the modified 48-well Boyden chamber. Secreted growth factors were quantified by ELISA. Results: CMF secrete autocrine or paracrine migration stimulating factors. Culture supernatant of CMF collected after 24, 48, and 72 h (=conditioned media) stimulated the migration of CMF ( 48.9 &#45 4.5; 60.3 &#45 5.3 and 67.8 &#45 6.4 cells/hpf, respectively). Heating of conditioned media to 95°C or addition of cycloheximide during the conditioning period abolished migration. Addition of PDGF-AB (2.5-50 ng/ml) or IGF-I (10-300 ng/ml) to CMF conditioned media further increased the migration of CMF to a maximum of 177 and 160%, respectively, when compared to the migration induced by conditioned medium alone. Addition of EGF (2.5-50 ng/ml) or TGF- &#103 1 (1-50 pg/ml) caused an increased CMF migration up to 139 and 128%, respectively. MCP-1 (5-50 ng/ml) and bFGF (10-200 ng/ml) had no effect on CMF migration. Conclusion: The growth factors PDGF-AB, IGF-I, EGF and TGF- &#103 1 stimulate the migration of CMF. However, factors secreted by CMF are essential for their ability to migrate in response to these growth factors. The identification of physiologically relevant migration inducing factors may help to elucidate the network of interactions and the complex mechanisms involved in intestinal wound healing or fibrosis.