Prognostic role of the stromal cell derived factor-1 in patients with hepatitis B virus-related acute-on-chronic liver failure

BACKGROUND Stromal cell derived factor-1(SDF-1)plays a pivotal role in the recruitment of stem cells to injured livers.However,the changes of SDF-l in patients with hepatitis B virus(HBV)-related acute-on-chronic liver failure(ACLF)have yet to be elucidated.AIM To study the SDF-1 changes in patients with HBV-related ACLF.METHODS 30 patients with HBV-related ACLF,27 patients with chronic hepatitis B and 20 healthy individuals are involved in our study.The SDF-l mRNA expression in liver tissue was detected by quantitative real-time polymerase chain reaction.Immunohistochemical staining was performed to illustrate the expression of SDFl,CXC receptor 4(CXCR4)and Ki67.The serum SDF-l concentrations were also detected by enzyme-linked immunosorbent assays.RESULTS The expression of SDF-1 mRNA from ACLF patients was remarkably higher than that from other patients(both P<0.05).The expression of SDF-l,CXCR4 and Ki67 from ACLF were the highest among the three groups(all P<0.01).The serum SDF-l levels in ACLF patients were significantly lower than that in other patients(both P<0.01).Moreover,in ACLF patients,the serum SDF-1 Levels were positively correlated with serum total bilirubin and international normalized ratio.In addition,the serum SDF-l levels in survival were significantly lower compared with the non-survivals(P<0.05).The area under the curve for the serum SDF-1 level in predicting 28-d mortality was 0.722(P<0.05).CONCLUSION This study provides the SDF-1 changes in patients with HBV-related ACLF.The SDF-1 Level at admission may serve as a promising prognostic marker for predicting short-term prognosis.

Microvesicles derived from mesenchymal stem cells inhibit acute respiratory distress syndrome-related pulmonary fibrosis in mouse partly through hepatocyte growth factor

BACKGROUND Pulmonary fibrosis is one of the main reasons for the high mortality rate among acute respiratory distress syndrome(ARDS)patients.Mesenchymal stromal cell-derived microvesicles(MSC-MVs)have been shown to exert antifibrotic effects in lung diseases.AIM To investigate the effects and mechanisms of MSC-MVs on pulmonary fibrosis in ARDS mouse models.METHODS MSC-MVs with low hepatocyte growth factor(HGF)expression(siHGF-MSC-MVs)were obtained via lentivirus transfection and used to establish the ARDS pulmonary fibrosis mouse model.Following intubation,respiratory mechanics-related indicators were measured via an experimental small animal lung function tester.Homing of MSC-MVs in lung tissues was investigated by near-infrared live imaging.Immunohistochemical,western blotting,ELISA and other methods were used to detect expression of pulmonary fibrosis-related proteins and to compare effects on pulmonary fibrosis and fibrosis-related indicators.RESULTS The MSC-MVs gradually migrated and homed to damaged lung tissues in the ARDS model mice.Treatment with MSC-MVs significantly reduced lung injury and pulmonary fibrosis scores.However,low expression of HGF(siHGF-MSC-MVs)significantly inhibited the effects of MSC-MVs(P<0.05).Compared with the ARDS pulmonary fibrosis group,the MSC-MVs group exhibited suppressed expression of type I collagen antigen,type III collagen antigen,and the proteins transforming growth factor-βandα-smooth muscle actin,whereas the siHGF-MVs group exhibited significantly increased expression of these proteins.In addition,pulmonary compliance and the pressure of oxygen/oxygen inhalation ratio were significantly lower in the MSC-MVs group,and the effects of the MSC-MVs were significantly inhibited by low HGF expression(all P<0.05).CONCLUSION MSC-MVs improved lung ventilation functions and inhibited pulmonary fibrosis in ARDS mice partly via HGF mRNA transfer.

Stromal cell derived factor-1 enhances bone marrow mononuclear cell migration in mice with acute liver failure

AIM： To evaluate the number of bone marrow mononuclear cells （BMMC） that are migrated to the liver following transplantation of murine BMMC into mice with acute liver injury.METHODS： BMMC were isolated from the bone marrow of mice in a lymphocyte separation medium and then labeled with PKH26. The labeled cells were subsequently infused into the caudal veins of BALB/c mice with hepatic injury induced by carbon tetrachloride and 2-acetylaminofluorene. Mice in experimental group were treated with stromal cell-derived factor-1 （SDF-1） which was injected intraperitoneally after trans- plantation of BMMC. Mice in control group were injected intraperitoneally with 0.1 mL of saline （0.9% NaCl） after transplantation of BMMC. After 2 wk, migration of the cells in experimental group was studied by fluorescence microscopy. The expression of proliferating cell nuclear antigen and albumin was quantified with manual methods in both groups. The serum transaminase levels at different time points were compared between the two groups.RESULTS： The labeled ＂cells＂ were found in the portal region and central veins of hepatic Iobules. The PKH26labeled cells appeared at an average frequency of 108 ± 8/high power field in the experiment group and 65 ± 8/high power field in the control group （P 〈 0.05）. The total number of positive cells was 29 ± 7/high power field in the experimental group and 13 ± 2/high power field in the control group. The albumin expression level was also higher in the experimental group than in the control group （29 ± 7 vs 13 ± 2, P 〈 0.05）. The total number of crossing points was 156 ± 5/high power field in the experimental group and 53 ± 5/high power field in the control group （P 〈 0.05）. The serum alanine aminotransferase levels in experimental and control groups were measured at different time points （120 ± 40 vs 118.50 ± 1.75, P 〉 0.05; 80.60 ± 6.50 vs 101.08 ± 5.67, P 〈 0.05; 50.74 ± 5.38 vs 80.47 ± 4.62, P 〈 0.05; 30.54 ± 2.70 vs 60.72 ± 4.37, P 〈 0.05; 30.77 ± 5.36 vs 40.47 ± 6.50, P 〈 0.05）. At the same time, the serum aspartate aminotransferase levels were measured in experimental and control groups at different time points （122.55 ± 1.46 vs 120.70 ± 4.22, P 〉 0.05; 54.26 ± 6.50 vs 98.70 ± 8.20, P 〈 0.05; 39.47 ± 5.39 vs 78.34 ± 4.50, P 〈 0.05; 28.94 ±2.70 vs 56.44 ± 4.28, P 〈 0.05; 30.77 ± 5.45 vs 42.50 ± 6.28, P 〈 0.05）.CONCLUSION： SDF-1 can promote the migration of BMMC to the liver of mice with acute liver failure.

Stromal cell-derived factor-1α promotes recruitment and differentiation of nucleus pulposus-derived stem cells

BACKGROUND Intervertebral disc(IVD) degeneration is a condition characterized by a reduction in the water and extracellular matrix content of the nucleus pulposus(NP) and is considered as one of the dominating contributing factors to low back pain. Recent evidence suggests that stromal cell-derived factor 1α(SDF-1α) and its receptor CX-C chemokine receptor type 4(CXCR4) direct the migration of stem cells associated with injury repair in different musculoskeletal tissues.AIM To investigate the effects of SDF-1α on recruitment and chondrogenic differentiation of nucleus pulposus-derived stem cells(NPSCs).METHODS We performed real-time RT-PCR and enzyme-linked immunosorbent assay to examine the expression of SDF-1α in nucleus pulposus cells after treatment with pro-inflammatory cytokines in vitro. An animal model of IVD degeneration was established using annular fibrosus puncture in rat coccygeal discs. Tissue samples were collected from normal control and degeneration groups.Differences in the expression of SDF-1α between the normal and degenerative IVDs were analyzed by immunohistochemistry. The migration capacity of NPSCs induced by SDF-1α was evaluated using wound healing and transwell migration assays. To determine the effect of SDF-1α on chondrogenic differentiation of NPSCs, we conducted cell micromass culture and examined the expression levels of Sox-9, aggrecan, and collagen II. Moreover, the roles of SDF-1/CXCR4 axis in the migration and chondrogenesis differentiation of NPSCs were analyzed by immunofluorescence, immunoblotting, and real-time RT-PCR.RESULTS SDF-1α was significantly upregulated in the native IVD cells cultured in vitro with pro-inflammatory cytokines, such as interleukin-1β and tumor necrosis factor-α, mimicking the degenerative settings. Immunohistochemical staining showed that the level of SDF-1α was also significantly higher in the degenerative group than in the normal group. SDF-1α enhanced the migration capacity of NPSCs in a dose-dependent manner. In addition, SDF-1α induced chondrogenic differentiation of NPSCs, as evidenced by the increased expression of chondrogenic markers using histological and immunoblotting analyses. Realtime RT-PCR, immunoblotting, and immunofluorescence showed that SDF-1αnot only increased CXCR4 expression but also stimulated translocation of CXCR4 from the cytoplasm to membrane, accompanied by cytoskeletal rearrangement.Furthermore, blocking CXCR4 with AMD3100 effectively suppressed the SDF-1α-induced migration and differentiation capacities of NPSCs.CONCLUSION These findings demonstrate that SDF-1α has the potential to enhance recruitment and chondrogenic differentiation of NPSCs via SDF-1/CXCR4 chemotaxis signals that contribute to IVD regeneration.

Neuroprotective effects of exogenous brain-derived neurotrophic factor on amyloid-beta 1-40-induced retinal degeneration

Amyloid-beta(Aβ)-related alterations,similar to those found in the brains of patients with Alzheimer's disease,have been observed in the retina of patients with glaucoma.Decreased levels of brain-derived neurotrophic factor(BDNF)are believed to be associated with the neurotoxic effects of Aβpeptide.To investigate the mechanism underlying the neuroprotective effects of BDNF on Aβ_(1-40)-induced retinal injury in Sprague-Dawley rats,we treated rats by intravitreal administration of phosphate-buffered saline(control),Aβ_(1-40)(5 nM),or Aβ_(1-40)(5 nM)combined with BDNF(1μg/mL).We found that intravitreal administration of Aβ_(1-40)induced retinal ganglion cell apoptosis.Fluoro-Gold staining showed a significantly lower number of retinal ganglion cells in the Aβ_(1-40)group than in the control and BDNF groups.In the Aβ_(1-40)group,low number of RGCs was associated with increased caspase-3 expression and reduced TrkB and ERK1/2 expression.BDNF abolished Aβ_(1-40)-induced increase in the expression of caspase-3 at the gene and protein levels in the retina and upregulated TrkB and ERK1/2 expression.These findings suggest that treatment with BDNF prevents RGC apoptosis induced by Aβ_(1-40)by activating the BDNF-TrkB signaling pathway in rats.

Effect of stromal cell-derived factor-1/CXCR4 axis in neural stem cell transplantation for Parkinson’s disease

Previous studies have shown that neural stem cell transplantation has the potential to treat Parkinson’s disease,but its specific mechanism of action is still unclear.Stromal cell-derived factor-1 and its receptor,chemokine receptor 4(CXCR4),are important regulators of cell migration.We speculated that the CXCR4/stromal cell-derived factor 1 axis may be involved in the therapeutic effect of neural stem cell transplantation in the treatment of Parkinson’s disease.A Parkinson’s disease rat model was injected with 6-hydroxydopamine via the right ascending nigrostriatal dopaminergic pathway,and then treated with 5μL of neural stem cell suspension(1.5×104/L)in the right substantia nigra.Rats were intraperitoneally injected once daily for 3 days with 1.25 mL/kg of the CXCR4 antagonist AMD3100 to observe changes after neural stem cell transplantation.Parkinson-like behavior in rats was detected using apomorphine-induced rotation.Immunofluorescence staining was used to determine the immunoreactivity of tyrosine hydroxylase,CXCR4,and stromal cell-derived factor-1 in the brain.Using quantitative real-time polymerase chain reaction,the mRNA expression of stromal cell-derived factor-1 and CXCR4 in the right substantia nigra were measured.In addition,western blot assays were performed to analyze the protein expression of stromal cell-derived factor-1 and CXCR4.Our results demonstrated that neural stem cell transplantation noticeably reduced apomorphine-induced rotation,increased the mRNA and protein expression of stromal cell-derived factor-1 and CXCR4 in the right substantia nigra,and enhanced the immunoreactivity of tyrosine hydroxylase,CXCR4,and stromal cell-derived factor-1 in the brain.Injection of AMD3100 inhibited the aforementioned effects.These findings suggest that the stromal cell-derived factor-1/CXCR4 axis may play a significant role in the therapeutic effect of neural stem cell transplantation in a rat model of Parkinson’s disease.This study was approved by the Animal Care and Use Committee of Kunming Medical University,China(approval No.SYXKK2015-0002)on April 1,2014.

Upregulation of stromal cell-derived factor-1 alpha/CXCR4 axis-induced migration of human neural progenitors by tumor necrosis factor-alpha and interleukin-8

BACKGROUND： Studies of several animal models of central nervous system diseases have shown that neural progenitor cells （NPCs） can migrate to injured tissues. Stromal cell-derived factor 1 alpha （SDF-la）, and its primary physiological receptor CXCR4, have been shown to contribute to this process. OBJECTIVE： To investigate migration efficacy of human NPCs toward a SDF-1α gradient, and the regulatory roles of tumor necrosis factor-α （TNF-α） and interleukin-8 （IL-8） in SDF-1α/CXCR4 axis-induced migration of NPCs. DESIGN, TIME AND SETTING： An in vitro, randomized, controlled, cellular and molecular biology study was performed at the Laboratory of Department of Cell Biology, Medical College of Soochow University between October 2005 and November 2007. MATERIALS： SDF-1α and mouse anti-human CXCR4 fusion antibody were purchased from R＆D Systems, USA. TNF-αwas purchased from Biomyx Technology, USA and IL-8 was kindly provided by the Biotechnology Research Institute of Soochow University. METHODS： NPCs isolated from forebrain tissue of 9 to 10-week-old human fetuses were cultured in vitro. The cells were incubated with 0, 20, and 40 ng/mL TNF-α, or 0, 20, and 40 ng/mL IL-8, for 48 hours prior to migration assay. For antibody-blocking experiments, cells were further pretreated with 0, 20, and 40 μg/mL mouse anti-human CXCR4 fusion antibody for 2 hours. Subsequently, the transwell assay and CXCR4 blockade experiments were performed to evaluate migration of human NPCs toward a SDF-1α gradient. Serum-free culture medium without SDF-1α served as the negative control. MAIN OUTCOME MEASURES： The transwell assay was performed to evaluate migration of human NPCs toward a SDF-1α gradient, which was blocked by fusion antibody against CXCR4. In addition, CXCR4 expression in human NPCs stimulated by TNF-α and IL-8 was measured by flow cytometry. RESULTS： Results from the transwell assay demonstrated that SDF-1α was a strong chemoattractant for human NPCs （P 〈 0.01）, and 20 ng/mL produced the highest levels of migration. Anti-human CXCR4 fusion antibody significantly blocked the chemotactic effect （P 〈 0.05）. Flow cytometry results showed that treatment with TNF-α and IL-8 resulted in increased CXCR4 expression and greater chemotaxis efficiency of NPCs towards SDF-1α（P 〈 0.01）. CONCLUSION： These results demonstrated that SDF-la significantly attracted NPCs in vitro, and neutralizing anti-CXCR4 antibody could block part of this chemotactic function. TNF-α and IL-8 increased chemotaxis efficiency of NPCs towards the SDF-1αgradient by upregulating CXCR4 expression in NPCs.

Emodin and baicalein inhibit pancreatic stromal derived factor-1 expression in rats with acute pancreatitis

BACKGROUND: Stromal derived factor-1 (SDF-1) is an efficacious leukocyte chemoattractant, which can attract lymphocytes and mononuclear cells from bloodstream into the site of inflammation. Emodin., an anthraquinone derivative from Radix et Rhizoma Rhei, and baicalein, a flavone from Scutellaria baicalensis Georgi, both have been reported to possess anti-inflammatory activities. The expression pattern of SDF-1 in experimental acute pancreatitis (AP) and the effect of emodin or baicalein on that are not well defined. The present study aimed to investigate the effects of emodin and baicalein on pancreatic myeloperoxidase (MPO) activity (reflecting leukocyte sequestration) and cytokine production, as well as tissue SDF-1 expression in the setting of AP. METHODS: A :rat model of AP was induced by administration (of 5% sodium taurocholate through the biliopancreatic duct. The level of tumor necrosis factor-a (TNF-alpha), interleukin-6 (IL-6) and MPO in the pancreas, and serum amylase were tested by immunohistochemistry, ELISA and chromatometry. The expressions of SDF-1 alpha and SDF-1 beta were detected by real-time PCR, Western blotting, and immunohistochemistry. RESULT: Combination of emodin and baicalein significantly reduced pancreatic TNIP-alpha, IL-6 and MPO, and also inhibited pancreatic SDF-1 expression. CONCLUSIONS: The inhibition of SDF-1 expression by emodin and baicalein might contribute, in part at least, to the amelioration of pancreatic inflammation. The present study also shows benefits of simultaneous treatment of AP.

Neuron-like differentiation of adult rat bone marrow stromal cells induced by transforming growth factor-beta and brain-derived neurotrophic factor

BACKGROUND： It has been demonstrated that transforming growth factor-β （TGF-β） and brain- derived neurotrophic factor （BDNF） can induce stem cell differentiation into neuron-like cells. OBJECTIVE： To investigate the efficacy of TGF-β and BDNF at inducing the differentiation of adult rat bone marrow stromal cells （BMSCs） into neuron-like cells, both in combination or alone. DESIGN, TIME AND SETTING： A comparative observation experiment was performed at the Department of Orthopedics, First Affiliated Hospital of Liaoning Medical University between October 2007 and January 2008. MATERIALS： TGF-~ and BDNF were purchased from Sigma, USA; mouse anti-rat neuron specific enolase, neurofilament and glial fibrillary acidic protein were purchased from Beijing HMHL Biochem Ltd., China. METHODS： BMSCs were isolated from rats aged 4 weeks and incubated with TGF-β（1μ g/L） and/or BDNF （50 μ g/mL）. MAIN OUTCOME MEASURES： Expression of neuron-specific enolase, neurofilament and glial fibrillary acidic protein were determined by immunocytochemistry. RESULTS： BMSCs differentiated into neuron-like cells following induction of TGF-β and BDNF, and expressed both neuron-specific enolase and neurofilament. The percent of positive cells was significantly greater in the combination group than those induced with TGF-β or BDNF alone （P 〈 0.01）. CONCLUSION： Treatment of BMSCs with a combination of TGF-β and BDNF induced differentiation into neuron-like cells, with the induction being significantly greater than with TGF-β or BDNF alone.

KIT and platelet-derived growth factor receptor α wild-type gastrointestinal stromal tumor associated with neurofibromatosis type 1: Two case reports

BACKGROUND Gastrointestinal stromal tumors(GISTs) associated with neurofibromatosis are uncommon compared to their gastrointestinal counterparts. Patients with neurofibromatosis type 1(NF-1) have an increased risk of developing gastrointestinal tumors, including rare types such as GIST.CASE SUMMARY A 60-year-old male Chinese patient was diagnosed with NF-1 10 years ago and presented with upper abdominal discomfort and black stools. Endoscopic ultrasonography and an enhanced abdominal computed tomography scan revealed a mass located 4 cm from the muscular layer of the descending duodenum. A 59-year-old Chinese woman who was diagnosed with NF-1 25 years ago presented with sudden unconsciousness and black stools. Multiple masses in the duodenum were noted by echogastroscopy and an enhanced abdominal computed tomography scan. Both patients presented with cutaneous neurofibromas. The histologic examination of tumors from both patients revealed spindle cells and low mitotic activity. Immunohistochemically, the tumor cells showed strong positivity for KIT(CD117), DOG-1, CD34, and Dehydrogenase Complex Subunit B, and negativity for SMA, desmin, S-100, and β-catenin. None of the six tumors from two patients had KIT exon 9, 11, 13, or 17 or platelet-derived growth factor receptor α exon 12 or 18 mutation, which is a typical finding for sporadic GISTs. None of the six tumors from the two patients had a BRAFV600 E mutation. The patients were alive and well during the follow-up period(range:0.6-5 yr).CONCLUSION There have been only a few previous reports of GISTs associated with NF-1.Although GISTs associated with NF-1 have morphologic and immunohistochemical similarities with GISTs, the pathogenesis, incidence,genetic background, and prognosis are not completely known. A medical history of NF-1 in a patient who has gastrointestinal bleeding or anemia and an intraabdominal mass with nonspecific computed tomography features may help in diagnosing GIST by virtue of the well-known association of these two entities.Molecular genetic studies of cases indicated that GISTs in NF-1 patients have a different pathogenesis than sporadic GISTs.

Effects of interleukin-1α on platelet-derived growth factor release from bovine cerebral microvascular endothelial cells

The present study showed that, interleukin-1α (IL-1α) stimulateal cultured bovine cerebral microvascular endothelial cells (BCMEC) releasing growth factor which promoted bovine cerebral microvascular smooth muscle cells (BCMSMC) proliferation in a dose-dependent manner. The mitogenic activity in conditioned medium of BCMEC stimulated by IL-1α was neutralized significantly by the antibody to platelet-derived growth factor (PDGF). Imperatorin (Imp), iso-Imperatorin (iso Imp) and 6-(α,α-phenylacetylpiperazinyl)phenyl-5-methyl-4,5-dihydro-3 (2H)-pyridazinone (PMDP) did not affect the releasing of PDGF from IL-1α stimulated BCMEC, but inhibited the promotion of PDGF on the proliferation of BCMSMC. We concluded that the promotion of IL-1α on the proliferation of BCMSMC should be mediated by some growth factors, such as PDGF.

NONHSAT248596.1内源性竞争miR-146a-5p调控骨关节炎软骨退变的机制

背景:目前已有针对lncRNA\miRNA\mRNA的共表达网络对骨关节炎发生发展调控机制的研究,课题组前期研究已通过数据库筛选出符合条件的NONHSAT248596.1和miR-146a-5p,尚缺乏体内实验来验证上述调控机制。目的:探究NONHSAT248596.1在基质细胞衍生因子1/4型趋化因子受体轴介导体内骨关节炎软骨退变进程中对miR-146a-5p发挥的竞争性内源性RNA调控作用。方法:取36只新西兰兔,通过向右侧后肢膝关节注射基质细胞衍生因子1溶液建立骨关节炎模型,采用随机数字表法分4组,lncRNA组、miRNA组、ceRNA组、对照组分别向造模膝关节内注射NONHSAT248596.1过表达的慢病毒载体、miR-146a-5p过表达的慢病毒载体、miR-146a-5p+NONHSAT248596.1过表达的慢病毒载体及空慢病毒载体。造模第4,8,12周,取膝关节软骨组织和软骨下骨组织进行相关检测。结果与结论:①苏木精-伊红与番红O固绿染色显示,4组软骨组织都有不同程度的退变表现,造模第4周时,lncRNA组软骨组织中的软骨细胞肿胀、细胞极性消失,细胞外基质破坏,出现表层糜烂、裂缝形成和软骨组织局部或全层缺失,并随时间延长软骨损伤程度逐渐加重,4组中miRNA组关节软骨炎症进展最缓慢;②qRT-PCR检测显示,相同时间点下,lncRNA组软骨组织中NONHSAT248596.1、4型趋化因子受体、基质金属蛋白酶3,9及13的mRNA表达量高于其他3组(P<0.05),miR-146a-5p、聚集蛋白聚糖及Ⅱ型胶原的mRNA表达量低于其他3组(P<0.05);造模后第8,12周,miRNA组软骨组织中的NONHSAT248596.1、4型趋化因子受体、基质金属蛋白酶3,9及13的mRNA表达量低于ceRNA组、对照组(P<0.05),miR-146a-5p、聚集蛋白聚糖及Ⅱ型胶原的mRNA表达量高于ceRNA组、对照组(P<0.05);③Western Blot检测显示,相同时间点下,lncRNA组软骨组织中的聚集蛋白聚糖及Ⅱ型胶原蛋白表达量始终低于其他3组(P<0.05);miRNA组造模后第8,12周软骨组织中的聚集蛋白聚糖及Ⅱ型胶原蛋白表达量高于ceRNA组、对照组(P<0.05);④结果表明,miR-146a-5p作为NONHSAT248596.1的作用靶点会受到其竞争性内源性RNA的作用造成活性被抑制,NONHSAT248596.1作用于miR-146a-5p后调控基质细胞衍生因子1/4型趋化因子受体轴,影响骨关节炎软骨组织中基质金属蛋白、Ⅱ型胶原、聚集蛋白聚糖的表达,造成细胞外基质的降解及蛋白多糖的丢失。

胃癌组织中SDF-1、HER2及Slug表达与患者临床病理特征的相关性

目的分析基质细胞衍生因子1(SDF-1)、人表皮生长因子受体2(HER2)、锌指转录因子(Slug)在胃癌组织内的表达及与患者临床病理特征间的关系。方法选取73例胃癌患者,采集其癌组织与癌旁正常组织,以免疫组织化学法检测对比两者SDF-1、HER2及Slug的表达差异;另收集患者的年龄等资料,统计分析SDF-1、HER2及Slug表达与胃癌患者各项临床病理特征间的联系。结果癌组织的SDF-1、HER2、Slug阳性表达率高于癌旁正常组织,差异有统计学意义(P<0.05)。SDF-1、HER2、Slug阳性表达与胃癌患者的年龄、性别无关(P>0.05),与患者的临床分期、淋巴结转移、分化程度有关(P<0.05)。结论SDF-1、HER2、Slug在胃癌组织内呈异常高表达,且其参与胃癌的侵袭、发展过程。

Intrastriatal glial cell line-derived neurotrophic factors for protecting dopaminergic neurons in the substantia nigra of mice with Parkinson disease

BACKGROUND： Substantia nigra is deep in position and limited in range, the glial cell line-derived neurotrophic factor （GDNF） injection directly into substantia nigra has relatively greater damages with higher difficulty. GDNF injection into striatum, the target area of dopaminergic neuron, may protect the dopaminergic neurons in the compact part of substantia nigra through retrograde transport. OBJECTIVE： To investigate the protective effect of intrastriatal GDNF on dopaminergic neurons in the substantia nigra of mice with Parkinson disease （PD）, and analyze the action pathway. DESIGN： A controlled observation. SETTING： Neurobiological Laboratory of Xuzhou Medical College. MATERIALS： Twenty-four male Kunming mice of 7 - 8 weeks old were used. GDNF, 1-methy1-4-pheny1-1,2,3,6-tetrahydropyridine （MPTP） were purchased from Sigma Company （USA）; LEICAQWin image processing and analytical system. METHODS： The experiments were carded out in the Neurobiological Laboratory of Xuzhou Medical College from September 2005 to October 2006. The PD models were established in adult KunMing mice by intraperitoneal injection of MPTP. The model mice were were randomly divided into four groups with 6 mice in each group： GDNF 4-day group, phosphate buffer solution （PSB） 4-day group, GDNF 6-day group and PSB 6-day group. Mice in the GDNF 4 and 6-day groups were administrated with 1 μ L GDNF solution （20 μ g/L, dispensed with 0.01 mol/L PBS） injected into right striatum at 4 and 6 days after model establishment. Mice in the PSB 4 and 6-day groups were administrated with 0.01 mol/L PBS of the same volume to the same injection at corresponding time points. ② On the 12^th day after model establishment, the midbrain tissue section of each mice was divided into 3 areas from rostral to caudal sides. The positive neurons of tyroxine hydroxylase （TH） and calcium binding protein （CB） with obvious nucleolus and clear outline were randomly selected for the measurement, and the number of positive neurons in unit area was counted. MAIN OUTCOME MEASURES： Number of positive neurons of TH and CB in midbrain substantia nigra of mice in each group. RESULTS： All the 24 mice were involved in the analysis of results. The numbers of TH^＋ and CB^＋ neurons in the GDNF 4-day group （54.33±6.92, 46.33±5.54） were obviously more than those in the PBS 4-day group （27.67±5.01, 21.50±5.96, P 〈 0.01）. The numbers of TH^＋ and CB^＋ neurons in the GDNF 6-day group （75.67±5.39, 69.67±8.69） were obviously more than those in the PBS 6-day group （27.17±4.50, 21.33 ±5.72, P 〈 0.01） and those in the GDNF 4-day group （P 〈 0.01 ）. CONCLUSION： Intrastriatal GDNF can protect dopaminergic neurons in substantia nigra of PD mice, and it may be related to the increase of CB expression.

槲皮素调节SDF-1/CXCR4信号轴对早发性卵巢功能不全大鼠卵巢结构及功能的影响

目的研究槲皮素通过调节基质细胞衍生因子(SDF-1)/CXC趋化因子受体4(CXCR4)信号通路对早发性卵巢功能不全(POI)大鼠卵巢结构及功能的影响。方法通过注射环磷酰胺构建POI大鼠模型,将大鼠随机分为对照组、模型组、槲皮素低剂量组(25.0 mg/kg)、槲皮素高剂量组(100 mg/kg)、槲皮素高剂量+CTCE-0214组(100 mg/kg槲皮素+10.0 mg/kg CTCE-0214),对照组大鼠给予等量0.9%氯化钠溶液。ELISA试剂盒检测大鼠血清中促卵泡激素(FSH)、雌二醇(E_(2))、抗苗勒管激素(AMH)、超氧化物歧化酶(SOD)、丙二醛(MDA)的表达;计算5组大鼠的卵巢指数;HE染色检测大鼠卵巢组织病理损伤并进行卵泡计数;免疫组化法检测卵巢雌激素受体(ER)蛋白的表达;Western blot检测大鼠卵巢组织中SDF-1、CXCR4蛋白表达。结果与对照组比较,模型组大鼠血清中FSH、MDA水平增加,E_(2)、AMH、SOD水平减少,卵巢指数降低,卵巢组织中成熟卵泡数目降低,ER蛋白表达减少,卵巢组织中闭锁卵泡数量、SDF-1、CXCR4蛋白表达增加(P<0.05)。与模型组比较,槲皮素低剂量组和槲皮素高剂量组大鼠血清中FSH、MDA水平降低,E_(2)、AMH、SOD水平升高,卵巢指数增加,卵巢组织中成熟卵泡数目、ER蛋白表达增加,卵巢组织中闭锁卵泡数量、SDF-1、CXCR4蛋白表达减少(P<0.05)。槲皮素高剂量+CTCE-0214组与槲皮素高剂量组比较,POI大鼠卵巢结构受损加重,卵巢功能受到影响,差异有统计学意义(P<0.05)。结论槲皮素可能通过抑制SDF-1/CXCR4信号通路对早发性卵巢功能不全大鼠的卵巢结构及功能发挥保护作用。

吴茱萸碱调节SDF-1α/CXCR4信号通路对颅内动脉瘤血管平滑肌细胞增殖和凋亡的影响

目的探究吴茱萸碱通过调节基质细胞衍生因子1α(SDF-1α)/CXC趋化因子受体4(CXCR4)信号通路对颅内动脉瘤(IA)血管平滑肌细胞(VSMCs)的增殖和凋亡的作用。方法24只小鼠随机分为对照组和IA组,每组12只。IA组通过定位手术注射弹性蛋白酶制造IA模型小鼠,然后通过HE染色观察动脉组织变化。随后将小鼠主动脉血管平滑肌细胞(MOVAS)先用MTT法检测吴茱萸碱浓度对细胞的活性影响,然后将MOVAS分为ctrl组、Model组(H_(2)O_(2)诱导损伤组)、低浓度吴茱萸碱组(0.50μmol/L)、高浓度吴茱萸碱组(1.00μmol/L)、高浓度吴茱萸碱+CTCE-0214组(1.00μmol/L吴茱萸碱+10 mg/kg SDF-1α/CXCR4激活剂)。CCK-8试剂盒检测细胞活性;流式细胞术检测细胞凋亡;Western blot检测SDF-1α、CXCR4、BAX、增殖细胞核抗原(PCNA)、平滑肌22α(SM22α)和平滑肌α肌动蛋白(α-SMA)的蛋白表达。结果与对照组正常动脉组织比较,IA组的IA组织出现明显的病理变化,损伤严重。在MOVAS细胞实验中,与ctrl组比较,Model组的细胞凋亡率、SDF-1α、CXCR4、BAX蛋白表达增加,而细胞存活率、PCNA、SM22α、α-SMA含量降低(P<0.05)。与Model组比较,低浓度吴茱萸碱组、高浓度吴茱萸碱组的细胞凋亡率、SDF-1α、CXCR4、BAX蛋白表达降低,而细胞存活率、PCNA、SM22α、α-SMA含量升高(P<0.05);与高浓度吴茱萸碱组比较,高浓度吴茱萸碱+CTCE-0214组细胞凋亡率、SDF-1α、CXCR4、BAX蛋白表达升高,而细胞存活率、PCNA、SM22α、α-SMA含量降低(P<0.05)。结论吴茱萸碱可能通过抑制SDF-1α/CXCR4信号通路进而抑制颅内动脉瘤血管平滑肌细胞的凋亡,促进其增殖。

骨髓增生异常综合征患者血清TGF-β1、SDF-1水平变化与预后的关系

目的:探讨骨髓增生异常综合征(MDS)患者血清转化生长因子β1(TGF-β1)和基质细胞衍生因子1(SDF-1)表达水平变化与预后的关系。方法:研究组选取92例在本院确诊治疗的MDS患者,对照组选取92例进行健康体检的志愿者(2021年1月至2023年6月)。采用酶联免疫吸附(ELISA)法测定研究组与对照组血清TGF-β1、SDF-1的水平变化;多因素Logistic回归探讨预后不良的影响因素;采用受试者工作特征(ROC)曲线评估TGF-β1、SDF-1对患者预后的预测价值。结果:与对照组相比,研究组血清TGF-β1水平显著下降,SDF-1水平显著上升(均P<0.05)。且随着IPSS评分等级的增加,TGF-β1的水平呈显著下降趋势,SDF-1的水平呈显著上升趋势(P<0.05)。预后不良组TGF-β1水平显著低于预后良好组,SDF-1水平显著高于预后良好组(均P<0.05)。多因素Logistic回归分析结果显示,TGF-β1低表达水平为MDS患者预后不良的危险因素,SDF-1高表达水平为MDS患者预后不良的危险因素(P<0.05)。根据ROC曲线得知,血清TGF-β1、SDF-1预测MDS患者预后的AUC分别为0.754、0.798,两者联合预测的AUC显著大于TGF-β1、SDF-1单独预测的AUC(Z_(TGF-β1)=2.733、Z_(SDF-1)=2.162,均P<0.05)。结论:MDS患者血清TGF-β1表达水平降低,而SDF-1表达水平升高,二者是影响MDS患者预后的因素,联合TGF-β1、SDF-1表达水平预测MDS患者预后的价值较高。

西格列汀激活基质细胞衍生因子-1/CXC趋化因子受体4信号通路对脂多糖诱导的人牙周膜干细胞增殖、凋亡、炎症和成骨分化的影响

目的 探讨西格列汀对脂多糖(LPS)诱导的炎症微环境下人牙周膜干细胞(hPDLSCs)增殖、凋亡、炎症和成骨分化的影响及分子机制。方法 体外培养hPDLSCs,用不同浓度的西格列汀处理后检测细胞活力,以确定后续西格列汀实验浓度。采用1μg/mL LPS刺激诱导24 h建立hPDLSCs炎症模型并分为空白组、对照组、西格列汀低浓度组(0.5μmol/L)、西格列汀中浓度组(1μmol/L)、西格列汀高浓度组(2μmol/L)、西格列汀高浓度+基质细胞衍生因子-1 (SDF-1)/CXC趋化因子受体4 (CXCR4)通路抑制剂(AMD3100)组(2μmol/L+10μg/mL)。细胞计数试剂盒-8检测培养24、48、72 h后的hPDLSCs增殖活性;流式细胞术检测培养72 h后hPDLSCs凋亡情况;诱导成骨分化21 d后茜素红染色检测hPDLSCs成骨分化能力,试剂盒测定hPDLSCs中碱性磷酸酶(ALP)活性;酶联免疫吸附检测hPDLSCs培养上清液中炎症因子肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6水平;实时荧光定量聚合酶链反应(RT-qPCR)检测hPDLSCs中成骨分化相关基因Runt相关转录因子2 (RUNX2)、骨钙素(OCN)、骨桥蛋白(OPN)及SDF-1和CXCR4 mRNA表达;Western blot检测hPDLSCs中SDF-1、CXCR4蛋白表达。结果 与空白组比较,对照组hPDLSCs增殖活性、矿化结节数量、染色强度、ALP活性和RUNX2、OCN、OPN mRNA及SDF-1、CXCR4 mRNA和蛋白表达水平显著降低,凋亡率、TNF-α、IL-1β、IL-6水平显著升高(P<0.05);与对照组比较,西格列汀低、中、高浓度组hPDLSCs增殖活性、矿化结节数量、染色强度、ALP活性和RUNX2、OCN、OPN mRNA及SDF-1、CXCR4 mRNA和蛋白表达水平依次升高,凋亡率、TNF-α、IL-1β、IL-6水平依次降低(P<0.05);AMD3100可部分逆转高浓度西格列汀对LPS诱导的hPDLSCs的作用效果(P<0.05)。结论 西格列汀可能通过激活SDF-1/CXCR4信号通路促进LPS诱导的炎症微环境下hPDLSCs的增殖和成骨分化,抑制hPDLSCs凋亡和炎症反应。

急性脑梗死患者静脉溶栓前后血清内皮素-1、基质细胞衍生因子-1变化与功能结局的关系

目的探讨急性脑梗死(Acute cerebral infarction,ACI)患者静脉溶栓前后血清内皮素-1(Endothelin-1,ET-1)、基质细胞衍生因子-1(Stromal cell derived factor 1,SDF-1)水平变化,分析ET-1、SDF-1与功能结局的关系。方法选取2021年2月-2023年2月大兴区人民医院收治的103例ACI患者。根据日常生活能力(Activity of daily living,ADL)量表将其分为功能结局良好组及功能结局不良组。检测患者溶栓前后血清ET-1和SDF-1水平。Logistic回归模型分析ACI功能结局的影响因素,Pearson相关性分析血清ET-1、SDF-1水平与ADL评分的相关性,根据受试者工作特征(Receiver operating characteristics,ROC)曲线分析血清ET-1、SDF-1水平对ACI功能结局的预测价值。结果ACI患者溶栓1周后血清ET-1水平降低,SDF-1水平升高(P<0.05);ACI患者功能结局不良发生率为39.81%;与功能结局良好组比较,功能结局不良组发病至溶栓时间、NIHSS评分、合并心房室颤占比、溶栓前、溶栓1周后血清ET-1水平升高,SDF-1水平降低(P<0.05);Logistic回归分析结果显示,发病至溶栓时间长、NIHSS评分高、溶栓1周后的血清ET-1高水平及SDF-1低水平是ACI功能结局不良的独立危险因素(P<0.05);Pearson相关性分析显示,ADL评分与溶栓1周后的血清ET-1水平呈正相关(r=0.563,P<0.05),与SDF-1呈负相关(r=-0.483,P<0.05);ROC分析结果显示,溶栓1周后的血清ET-1、SDF-1二者联合预测ACI功能结局的AUC高于各项单独检测(P<0.05)。结论ACI患者溶栓后血清ET-1水平降低,SDF-1水平升高,溶栓1周后血清ET-1、SDF-1水平与ADL评分均呈正相关,且对于预测ACI功能结局有较高的参考价值。

淫羊藿苷调节SDF-1/CXCR4信号通路对多囊卵巢综合征大鼠卵巢颗粒细胞凋亡的影响

目的探讨淫羊藿苷调节基质细胞衍生因子-1(SDF-1)/CXC型趋化因子受体4(CXCR4)信号轴对多囊卵巢综合征(PCOS)大鼠卵巢颗粒细胞凋亡的影响。方法将从PCOS模型组大鼠卵巢组织中成功分离的颗粒细胞分为PCOS组、淫羊藿苷低剂量组、淫羊藿苷中剂量组、淫羊藿苷高剂量组、重组大鼠SDF-1蛋白(Rr-SDF-1)组、淫羊藿苷高剂量+Rr-SDF-1组,另取从正常对照组大鼠卵巢组织中成功分离的颗粒细胞作为对照组。酶联免疫吸附试验(ELISA)检测卵巢颗粒细胞上清液中卵泡刺激素(FSH)、睾酮(T)、黄体生成素(LH)水平;CCK-8法、EdU染色检测颗粒细胞增殖;流式细胞术检测颗粒细胞凋亡;Western blot检测颗粒细胞中Bcl-2相关X蛋白(Bax)、裂解的胱天蛋白酶-3(Cleaved Caspase-3)、SDF-1、CXCR4蛋白表达。结果与对照组比较,PCOS组FSH水平、光密度(OD)_(450)值、EdU阳性细胞率降低,T、LH水平、细胞凋亡率、Bax、Cleaved Caspase-3、SDF-1、CXCR4蛋白表达水平升高(P<0.05)。与PCOS组比较,淫羊藿苷低、中、高剂量组FSH水平、OD_(450)值、EdU阳性细胞率均依次升高,T、LH水平、细胞凋亡率、Bax、Cleaved Caspase-3、SDF-1、CXCR4蛋白表达水平均依次降低;Rr-SDF-1组对应指标变化趋势与上述相反(P<0.05)。与淫羊藿苷高剂量组比较,淫羊藿苷高剂量+Rr-SDF-1组FSH水平、OD_(450)值、EdU阳性细胞率降低,T、LH水平、细胞凋亡率、Bax、Cleaved Caspase-3、SDF-1、CXCR4蛋白表达水平升高(P<0.05)。结论淫羊藿苷可能通过抑制SDF-1/CXCR4信号通路抑制PCOS大鼠卵巢颗粒细胞凋亡。