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.

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.

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.

CXCR7/CXCL12 axis is involved in lymph node and liver metastasis of gastric carcinoma

AIM To investigate the role of CXC chemokine receptor (CXCR)-7 and CXCL12 in lymph node and liver metastasis of gastric carcinoma. METHODS In 160 cases of gastric cancer, the expression of CXCR7 and CXCL12 in tumor and matched tumoradjacent non-cancer tissues, in the lymph nodes around the stomach and in the liver was detected using immunohistochemistry to analyze the relationship between CXCR7/CXCL12 expression and clinicopathological features and to determine whether CXCR7 and CXCL12 constitute a biological axis to promote lymph node and liver metastasis of gastric cancer. Furthermore, the CXCR7 gene was silenced and overexpressed in human gastric cancer SGC-7901 cells, and cell proliferation, migration and invasiveness were measured by the MTT, wound healing and Transwell assays, respectively. RESULTS CXCR7 expression was up-regulated in gastric cancer tissues (P = 0.011). CXCR7/CXCL12 expression was significantly related to high tumor stage and lymph node (r = 0.338, P = 0.000) and liver metastasis (r = 0.629, P = 0.000). The expression of CXCL12 in lymph node and liver metastasis was higher than that in primary gastric cancer tissues (chi(2) = 6.669, P = 0.010; chi(2) = 25379, P = 0.000), and the expression of CXCL12 in lymph node and liver metastasis of gastric cancer was consistent with the positive expression of CXCR7 in primary gastric cancer (r = 0.338, P = 0.000; r = 0.629, P = 0.000). Overexpression of the CXCR7 gene promoted cell proliferation, migration and invasion. Silencing of the CXCR7 gene suppressed SGC-7901 cell proliferation, migration and invasion. Human gastric cancer cell lines expressed CXCR7 and showed vigorous proliferation and migratory responses to CXCL12. CONCLUSION The CXCR7/CXCL12 axis is involved in lymph node and liver metastasis of gastric cancer. CXCR7 is considered a potential therapeutic target for the treatment of gastric cancer.

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。

Growth and activation of PI-3K/PKB and Akt by stromal cell-derived factor 1a in endometrial carcinoma cells with expression of suppressor endoprotein PTEN

Background Mutation or deletion in the phosphatase and tensin homologue deleted on chromosome ten （PTEN） gene has been identified as an important cause of endometrial carcinoma; stromal cell derived factor-1α （SDF-1α） exerts growth-promoting effects on endometrial cancer cells through activation of the PI-3 kinase/Akt pathway and downstream effectors such as extracellular-responsive kinase （ERK）. In this study, a plasmid containing the PTEN gene was transfected into Ishikawa cells to investigate the difference in growth and signal transduction between Ishikawa-PTEN and Ishikawa cells after SDF-1α stimulation, and to study mechanisms of the involvement of PTEN protein in endometrial carcinoma development. Methods Ishikawa cells were transfected with a plasmid （pLXSN-PTEN） containing the PTEN gene and a plasmid （pLXSN-EGFP） with enhanced green fluorescent protein （EGFP）. Cells were then screened to obtain Ishikawa-PTEN cells and Ishikawa-neo cells that can both stably express PTEN protein and EGFP. Expression of PTEN protein, phosphorylation levels of AKT and ERK （pAKT and pERK） and growth differences in Ishikawa-PTEN, Ishikawa-neo and Ishikawa cells before and after SDF-1α stimulation were then determined by Western blots and MTT assays. Results Western blot analysis showed that Ishikawa cells produced PTEN after transfection with the PTEN gene. At 15 minutes after SDF-1α stimulation, the pAKT level of Ishikawa-PTEN cells was lower than that of Ishikawa-neo cells and Ishikawa cells. There was no significant difference in pERK levels among the three cell lines. The positive effect of SDF-1α on Ishikawa-PTEN cells growth was markedly less than the effect on Ishikawa-neo and Ishikawa cells. However, in the absence of SDF-1α stimulation （baseline）, the pAKT level in Ishikawa-PTEN cells was less than that in Ishikawa cells. There was a significant difference in growth between the Ishikawa-PTEN cells and the Ishikawa-neo cells. Conclusions PTEN gene transfection can regulate the level of pAKT but not pERK in Ishikawa-PTEN cells. PTEN protein may suppress the growth-promoting effect of SDF-1α on endometrial carcinoma by inhibiting the PI-3K/AKT signal transduction pathway.

SDF-1/CXCR4 expression in bladder cancer tissue and the correlation with negative costimulatory molecule PD-L1, cell apoptosis and invasion

Objective:To study the SDF-1/CXCR4 expression in bladder cancer tissue and the correlation with negative costimulatory molecule PD-L1, cell apoptosis and invasion.Methods: A total of 118 cases of bladder cancer tissue and para-carcinoma tissue surgically removed in our hospital between May 2014 and May 2016 were selected as the research samples, the RNA was extracted and then reverse-transcribed into cDNA, and the expression levels of SDF-1/CXCR4, PD-L1/PD-1, cell apoptosis-related molecules and cell invasion-related molecules were detected.Results: SDF-1 and CXCR4 mRNA expression in bladder cancer tissue were significantly higher than those in para-carcinoma tissue;PD-L1, PD-1, Rec1, Survivin, MRPS5, Nanog, BCAPP2Ac, TRPM8, TRPV2, ILK,β-catenin and GUGBP1 mRNA expression in bladder cancer tissue were significantly higher than those in para-carcinoma tissue and positively correlated with SDF-1 and CXCR4 mRNA expression.Conclusion:Highly expressed SDF-1/CXCR4 in bladder cancer tissue are closely related to the high expression of negative costimulatory molecule PD-L1, pro-proliferation molecules and pro-invasion molecules, and SDF-1/CXCR4 can promote the immune escape, proliferation and invasion of bladder cancer cells.

骨髓间充质干细胞与多发性骨髓瘤细胞共培养后CX43表达及SDF-1α分泌水平的变化及其意义

目的构建多发性骨髓瘤（MM）细胞与骨髓间充质干细胞（Msc）共培养体系，探讨共培养后MSC连接蛋白43（CX43）表达及基质细胞衍生因子（SDF）-1a分泌水平变化及其意义。方法用Westernblot、免疫荧光法检测MM细胞系及MM原代细胞CX43的表达及SDF-1a分泌水平。建立间接及直接共培养体系共培养MM细胞和MSC，然后用CD138磁珠法分离RPMI8226细胞及MSC。用实时定量PCR、Westernblot法检测共培养前后MSCCX43表达，免疫荧光法检测CX43分布；划痕实验检测共培养后MSC间间隙连接通讯（GJIC）变化；微孔隔离实验检测18a-甘草次酸（18a—GA）对MSC诱导的RPMI8226细胞迁移的影响。ELISA法检测共培养后的MSCSDF—1a分泌水平。结果MM细胞系RPMI8226、U266、1／3MM细胞及MM原代细胞CX43mRNA呈中、低度表达，XG-4、XG-7细胞不表达CX43。骨髓MSC高表达CX43。直接或间接共培养后骨髓MSC的CX43mRNA相对表达量明显提高，分别是单独培养时的1．36倍和2．10倍，Westernblot检测显示共培养后MSCCX43蛋白表达水平也上调，免疫荧光染色显示增高的CX43主要分布在胞质。划痕实验显示在MSC与RPMI8226直接共培养后荧光染料在细胞间扩散距离增加。MSC与RPMI8226细胞直接和间接共培养体系中，MSC培养上清SDF．1et水平分别为（373．02±10．11）和（309．714-10．71）pg／ml，高于共培养前[（237．84±9．23）pg／m1]（P〈0．01），该作用可被18a-GA抑制降为（126．01±4．80）和（106．99±3．39）pg／ml。18et—GA可抑制MSC诱导的RPMI8226细胞迁移，其作用前后RPMI8226细胞迁移率分别为（8．00--0．67）％及（4．82：50．19）％。结论MM细胞与MSC直接和间接共培养均可上调MSCCX43表达水平，并促进SDF—let的分泌。间隙连接阻断剂18et—GA可降低共培养体系中SDF—1a的分泌并抑制MSC诱导的MM细胞迁移。

Neuro-regenerative imidazole-functionalized GelMA hydrogel loaded with hAMSC and SDF-1α promote stem cell differentiation and repair focal brain injury

Brain tissues that are severely damaged by traumatic brain injury(TBI)is hardly regenerated,which leads to a cavity or a repair with glial scarring.Stem-cell therapy is one viable option to treat TBI-caused brain tissue damage,whose use is,whereas,limited by the low survival rate and differentiation efficiency of stem cells.To approach this problem,we developed an injectable hydrogel using imidazole groups-modified gelatin methacrylate(GelMA-imid).In addition,polydopamine(PDA)nanoparticles were used as carrier for stromal-cell derived factor-1(SDF-1α).GelMA-imid hydrogel loaded with PDA@SDF-1αnanoparticles and human amniotic mesenchymal stromal cells(hAMSCs)were injected into the damaged area in an in-vivo cryogenic injury model in rats.The hydrogel had low module and its average pore size was 204.61±41.41 nm,which were suitable for the migration,proliferation and differentiation of stem cells.In-vitro cell scratch and differentiation assays showed that the imidazole groups and SDF-1αcould promote the migration of hAMSCs to injury site and their differentiation into nerve cells.The highest amount of nissl body was detected in the group of GelMA-imid/SDF-1α/hAMSCs hydrogel in the in-vivo model.Additionally,histological analysis showed that GelMA-imid/SDF-1α/hAMSCs hydrogel could facilitate the regeneration of regenerate endogenous nerve cells.In summary,the GelMA-imid/SDF-1α/hAMSCs hydrogel promoted homing and differentiation of hAMSCs into nerve cells,and showed great application potential for the physiological recovery of TBI.

Syndecan-4 functionalization of tissue regeneration scaffolds improves interaction with endothelial progenitor cells

Key to most implanted cell free scaffolds for tissue regeneration is the ability to sequester and retain undifferentiated mesenchymal stem cells at the repair site.In this report,syndecan-4,a heparan sulfate containing proteoglycan,was investigated as a unique molecule for use in scaffold functionalization.An electrospun hybrid scaffold comprised of poly(glycerol)sebacate(PGS),silk fibroin and type I collagen(PFC)was used as a model scaffold to develop a procedure and test the hypothesis that functionalization would result in increased scaffold binding of endothelial progenitor cells(EPCs).For these studies both Syndecan-4 and stromal derived factor-1a(SDF-1a)were used in functionalization PFC.Syndecan-4 functionalized PFC bound 4.8 fold more SDF-1a compared to nonfunctionalized PFC.Binding was specific as determined by heparin displacement studies.After culture for 7 days,significantly,more EPCs were detected on PFC scaffolds having both syndecan-4 and SDF-1a compared to scaffolds of PFC with only syndecan-4,or PFC adsorbed with SDF-1a,or PFC alone.Taken together,this study demonstrates that EPCs can be bound to and significantly expanded on PFC material through syndecan-4 mediated growth factor binding.Syndecan-4 with a multiplicity of binding sites has the potential to functionalize and expand stem cells on a variety of scaffold materials for use in tissue regeneration.