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.

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 regulates chondrogenic differentiation via activation of the Wnt/β-catenin pathway in mesenchymal stem cells

BACKGROUND Mesenchymal stem cells(MSCs)have been applied to treat degenerative articular diseases,and stromal cell-derived factor-1α(SDF-1α)may enhance their therapeutic efficacy.However,the regulatory effects of SDF-1αon cartilage differentiation remain largely unknown.Identifying the specific regulatory effects of SDF-1αon MSCs will provide a useful target for the treatment of degenerative articular diseases.AIM To explore the role and mechanism of SDF-1αin cartilage differentiation of MSCs and primary chondrocytes.METHODS The expression level of C-X-C chemokine receptor 4(CXCR4)in MSCs was assessed by immunofluorescence.MSCs treated with SDF-1αwere stained for alkaline phosphatase(ALP)and with Alcian blue to observe differentiation.Western blot analysis was used to examine the expression of SRY-box transcription factor 9,aggrecan,collagen II,runt-related transcription factor 2,collagen X,and matrix metalloproteinase(MMP)13 in untreated MSCs,of aggrecan,collagen II,collagen X,and MMP13 in SDF-1α-treated primary chondrocytes,of glycogen synthase kinase 3β(GSK3β)p-GSK3βandβ-catenin expression in SDF-1α-treated MSCs,and of aggrecan,collagen X,and MMP13 in SDF-1α-treated MSCs in the presence or absence of ICG-001(SDF-1αinhibitor).RESULTS Immunofluorescence showed CXCR4 expression in the membranes of MSCs.ALP stain was intensified in MSCs treated with SDF-1αfor 14 d.The SDF-1αtreatment promoted expression of collagen X and MMP13 during cartilage differentiation,whereas it had no effect on the expression of collagen II or aggrecan nor on the formation of cartilage matrix in MSCs.Further,those SDF-1α-mediated effects on MSCs were validated in primary chondrocytes.SDF-1αpromoted the expression of p-GSK3βandβ-catenin in MSCs.And,finally,inhibition of this pathway by ICG-001(5μmol/L)neutralized the SDF-1α-mediated up-regulation of collagen X and MMP13 expression in MSCs.CONCLUSION SDF-1αmay promote hypertrophic cartilage differentiation in MSCs by activating the Wnt/β-catenin pathway.These findings provide further evidence for the use of MSCs and SDF-1αin the treatment of cartilage degeneration and osteoarthritis.

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.

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。

兔肩袖腱骨愈合过程中基质细胞衍生因子1的表达及作用机制

背景:近年在腱骨损伤领域部分学者将基质细胞衍生因子1搭载在组织工程支架上用以促进腱骨愈合,取得了较好的成效,但在基质细胞衍生因子1促进腱骨愈合机制及自然愈合过程中其是否参与修复,目前尚未明确。目的:研究兔肩袖全层冈上肌断裂后腱骨愈合过程中基质细胞衍生因子1表达,及其在腱骨损伤时对干细胞的迁移作用和最佳体外促迁移浓度。方法:随机取成年新西兰大白兔18只建立肩袖损伤模型,另取3只为空白对照。于造模后3,5,7,14,21,28 d各处死3只并处死空白组兔,取腱骨连接处组织保存在-80℃冰箱。应用ELISA反应检测损伤后各时间点愈合处基质细胞衍生因子1表达。取幼兔股骨骨髓间充质干细胞分离培养鉴定,通过transwell实验验证基质细胞衍生因子1对干细胞的促迁移作用效果及体外促迁移最佳浓度,将培养到P3代的干细胞与Brdu共培养后注入兔耳缘静脉,通过免疫组化染色验证干细胞是否迁移至损伤处。结果与结论:①基质细胞衍生因子1在肩袖腱骨愈合过程呈双峰表达,于伤后3 d明显增高(P<0.01)随后下降,于伤后5 d达最低,后再次升高于伤后14 d达峰值(P<0.01),然后下降;②细胞免疫组化染色可见标记有Brdu的干细胞确有迁移至损伤处;③transwell实验结果表明60-80 ng/mL的基质细胞衍生因子1对干细胞促迁移效果最好,而200 ng/mL浓度反而会起到抑制迁移作用;④基质细胞衍生因子1参与了肩袖腱骨愈合的炎症反应期和增殖期的愈合过程,其作用机制可能为通过促进干细胞迁移至损伤处并分化为各类细胞促进修复,并且基质细胞衍生因子1的促迁移作用存在于一定浓度范围,超出范围则可能起到抑制作用。

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。

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。

腹部创伤感染病人血清基质金属蛋白酶9、基质细胞衍生因子1表达及其诊断价值

目的 探究腹部创伤感染病人血清基质金属蛋白酶(MMP)-9、基质细胞衍生因子(SDF)-1表达及其诊断价值。方法 2021年7月~2022年7月收治的腹部创伤感染病人100例,作为病例组,同期腹部创伤未发生感染的病人100例为对照组。采用酶联免疫吸附法(ELISA)检测血清MMP-9、SDF-1水平;Spearman法分析血清MMP-9、SDF-1与SIRS评分和住院时间的相关性。ROC曲线分析血清MMP-9、SDF-1对腹部创伤感染病人的诊断价值。配对样本t检验分析腹部创伤感染病人治疗前后血清MMP-9、SDF-1水平。多因素Logistic回归分析影响腹部创伤感染发生的因素。结果 病例组病人血清MMP-9、SDF-1水平、C-反应蛋白(CRP)、白细胞计数(WBC)、血清中肿瘤坏死因子(TNF)-α、合并糖尿病、创伤类型等与对照组比较,差异有统计学意义(P<0.05)。腹部创伤感染病人血清MMP-9水平分别与SIRS评分、CRP、WBC、TNF-α呈正相关性(r=0.505、0.471、0.423、0.416,P<0.05);血清SDF-1水平分别与SIRS评分、CRP、WBC、TNF-α也呈正相关性(r=0.469、0.439、0.518、0.469,P<0.05)。ROC分析显示,血清MMP-9、SDF-1预测腹部创伤发生感染的曲线下面积(AUC)分别为0.850(95%CI:0.799~0.902)、0.806(95%CI:0.746~0.866),二者联合检测的AUC为0.914(95%CI:0.876~0.951),高于MMP-9、SDF-1单独检测(Z=1.987、2.97,P<0.05)。治疗后血清MMP-9、SDF-1表达水平低于治疗前,差异有统计学意义(P<0.05)。Logistic回归分析显示,合并糖尿病、创伤类型、血清MMP-9、SDF-1水平是腹部创伤病人感染的危险因素(P<0.05)。结论 腹部创伤感染病人血清MMP-9、SDF-1水平升高,二者可能参与腹部创伤感染的发生发展,对腹部创伤感染的诊断具有临床意义。

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。

基质细胞衍生因子1在软骨和软骨下骨稳态中的作用

背景:骨性关节炎是一种以软骨退变和软骨下骨异常骨重塑为特征的退行性病变。近年来,许多研究表明基质细胞衍生因子1在骨性关节炎的病理进展中发挥关键作用,靶向调控基质细胞衍生因子1及其CXC趋化因子受体4型和CXC趋化因子受体7型组成的信号通路是预防和治疗骨性关节炎的新方法。目的:综述基质细胞衍生因子1参与调控软骨细胞、骨髓间充质干细胞、成骨细胞及破骨细胞增殖、分化及凋亡的作用,以及这些细胞相互作用,探究导致软骨退变、软骨下骨异常骨重塑而加速骨性关节炎病理进展的机制,以期为骨性关节炎的防治提供新的思路。方法:以“基质细胞衍生因子1,软骨,软骨细胞,软骨下骨,骨髓间充质干细胞,成骨细胞,破骨细胞,CXC趋化因子受体4型,CXC趋化因子受体7型”为中文检索词检索中国知网、万方和维普数据库,以“Stromal cell-derived factor 1,SDF-1,CXCL12,cartilage,chondrocyte,subchondral bone,mesenchymal stem cells,osteoblasts,osteoclasts,CXCR4,CXCR7”为英文检索词检索PubMed、Medline和Embase数据库,检索各数据库建库至2024年1月的相关文献,根据纳入和排除标准,最终纳入77篇文献进行归纳总结。结果与结论:①基质细胞衍生因子1调控软骨细胞、骨髓间充质干细胞、成骨细胞和破骨细胞迁移、增殖、分化和死亡,在维持软骨和软骨下骨稳态以及促进或抑制骨性关节炎软骨退变、软骨下骨异常骨重塑等病理过程中均发挥至关重要的作用,靶向调控基质细胞衍生因子1/CXC趋化因子受体4型/CXC趋化因子受体7型信号通路有望成为今后防治骨性关节炎研究的重点。②由于基质细胞衍生因子1亚型在组织之间表达量的差异,目前以基质细胞衍生因子1α研究最为广泛,基质细胞衍生因子1β和基质细胞衍生因子1γ的相关研究多集中在探索影响干细胞生物学行为、在调控软骨和软骨下骨稳态中的作用,与骨性关节炎的相关性尚不清楚。③基质细胞衍生因子1能够有效促进干细胞归巢至软骨损伤部位,并诱导其增殖、存活及成软骨分化和应用负载基质细胞衍生因子1的生物支架来改善软骨修复质量已成为软骨组织工程研究的热点;然而,已有研究表明基质细胞衍生因子1可促进骨髓间充质干细胞向肥大型软骨细胞分化,而新生软骨细胞肥大表型会造成软骨内骨形成,软骨细胞凋亡,整个组织发生血管化和骨化,影响最终软骨修复的质量;另外,不同支架与基质细胞衍生因子1组合在修复部分软骨损伤和全层软骨损伤时,再生组织不都是理想的透明软骨组织。因而,未来深入探寻基质细胞衍生因子1在干细胞生物学效应中的潜在机制以及基质细胞衍生因子1与支架组合在修复不同软骨缺损的最佳组合方式将有助于提升软骨修复质量。④目前有关CXC趋化因子受体4型拮抗剂的研究主要集中在AMD3100,T140和TN14003,且绝大部分处于基础实验阶段,有待临床转化。针对基质细胞衍生因子1/CXC趋化因子受体4型/CXC趋化因子受体7型信号通路开发的治疗药物、方法的安全性、有效性仍需大量的生物学和临床试验加以佐证。

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。

甲基莲心碱调节SDF-1/CXCR4信号通路对糖尿病肾病的影响

目的·探讨甲基莲心碱(neferine,Nef)对糖尿病肾病(diabetic nephropathy,DN)大鼠肾组织的作用及其相关机制。方法·采用高脂饲料喂食联合腹腔注射链脲佐菌素的方法构建DN模型大鼠,并将造模成功的大鼠随机分为DN组、Nef(低、中、高)剂量组、Nef高剂量+通路拮抗剂(AMD3100)组,每组10只。同时,选10只普通大鼠作为正常组。检测6组大鼠的空腹血糖(fasting blood glucose,FBG)、24 h尿蛋白、血清糖化血红蛋白(glycosylated hemoglobin,HbA1c)、血清肌酐(serum creatinine,Scr)、尿素氮(blood urea nitrogen,BUN)水平及肾指数。分别采用苏木精-伊红(hematoxylin-eosin,H-E)染色、马松(Masson)染色观察6组大鼠的肾组织的病理变化。采用硫代巴比妥酸(thiobarbituric acid,TBA)法检测肾组织丙二醛(malondialdehyde,MDA)含量,分别采用水溶性四氮唑(water soluble tetrazolium,WST-1)法、钼酸铵法检测肾组织超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)的活性。分别采用实时荧光定量PCR(quantitative real-time PCR,qPCR)和蛋白质印迹法(Western blotting)检测肾组织中基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)、CXC趋化因子受体4(CXC chemokine receptor 4,CXCR4)的mRNA以及蛋白表达。采用高糖(30 mmol/L葡萄糖)诱导大鼠肾小管上皮细胞NRK-52E,以建立DN细胞模型。将该细胞分为对照组、高糖(HG)组、HG+Nef(低、中、高)剂量组(即HG+Nef-L、M、H组)、HG+Nef-H+AMD3100组。分别采用WST-1法、钼酸铵法检测模型细胞中SOD、CAT活性,采用TBA法检测MDA含量,分别采用qPCR、Western blotting检测SDF-1、CXCR4的mRNA及蛋白表达,采用CCK-8法、流式细胞术检测细胞活力和凋亡率。结果·与DN组比较,Nef(低、中、高)剂量组和Nef高剂量+AMD3100组大鼠的FBG、24 h尿蛋白、HbA1c、Scr、BUN水平以及肾指数、MDA水平均较低,SDF-1、CXCR4的mRNA和蛋白表达以及SOD、CAT活性均较高(均P<0.05),肾组织病理损伤、纤维化程度有所减轻,且均呈剂量依赖性;AMD3100能减弱高剂量Nef对DN大鼠的肾保护作用。与HG组比较,HG+Nef-L、M、H组NRK-52E细胞的活力,SOD、CAT活性,SDF-1、CXCR4的mRNA和蛋白表达均较高,MDA含量及凋亡率均较低(均P<0.05);AMD3100可逆转Nef-H对NRK-52E细胞损伤的保护作用。结论·Nef可能通过激活SDF-1/CXCR4信号通路来控制DN大鼠的血糖水平并提高其抗氧化能力,从而发挥肾保护作用。

围手术期HIF-1α、ANGPTL2、SDF-1与大面积脑梗死去骨瓣减压术后病情转归的相关性分析

目的探讨围手术期缺氧诱导因子-1α(HIF-1α)、血管生成素样蛋白2(ANGPTL2)、基质细胞衍生因子-1(SDF-1)与大面积脑梗死(LHI)去骨瓣减压术后病情转归的相关性。方法选取2020年6月至2022年12月在医院进行去骨瓣减压术的158例LHI患者,根据入院时美国国立卫生研究院卒中量表(NIHSS)评分分为观察组(71例,>20分)和对照组(87例,5~20分),另根据术后90 d改良Rankin量表(mRS)评分分为不良组(64例,>2分)和良好组(94例,≤2分)2个亚组。比较两组术前血清HIF-1α、ANGPTL2、SDF-1水平,分析血清HIF-1α、ANGPTL2、SDF-1水平与NIHSS评分的相关性,比较术前、术后14 d 2个亚组血清HIF-1α、ANGPTL2、SDF-1水平,分析LHI患者去骨瓣减压术后病情转归的影响因素及预测价值。结果术前观察组血清HIF-1α、ANGPTL2、SDF-1水平较对照组高(P<0.05);术前血清HIF-1α、ANGPTL2、SDF-1水平与NIHSS评分均呈正相关(P<0.05);与良好组相比,术后14 d不良组血清HIF-1α、ANGPTL2、SDF-1水平较高(P<0.05);术后14 d血清HIF-1α、ANGPTL2、SDF-1水平增加是预后不良的危险因素(P<0.05);血清HIF-1α、ANGPTL2、SDF-1水平联合预测的曲线下面积大于各指标单一预测(P<0.05)。结论血清HIF-1α、ANGPTL2、SDF-1表达上调不仅参与LHI发病,还与患者病情严重性及预后密切相关,早期检测有望成为辅助判断LHI病情、预测预后的重要生化指标。

SDF-1对骨髓间充质干细胞迁移和细胞外基质形成的影响

目的探讨基质细胞衍生因子(stromal cell-derived factor-1,SDF-1)对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)迁移、胶原RNA表达及细胞膜片细胞外基质表达的影响。方法CCK-8和Transwell细胞迁移实验观察SDF-1对BMSCs增殖和迁移的影响;实时荧光定量PCR检测BMSCs胶原RNA的表达水平;蛋白质印迹法检测BMSCs细胞膜片胶原蛋白的表达水平;酶联免疫吸附试验检测BMSCs细胞膜片培养基上清透明质酸(HA)的表达水平。结果SDF-1趋化BMSCs迁移作用的强度与浓度相关,对细胞的增殖无明显影响;SDF-1降低BMSCsⅠ型胶原与Ⅲ型胶原RNA和蛋白相对表达量的比值,并显著升高BMSCs细胞膜片透明质酸表达水平(P<0.05)。结论SDF-1在适宜的浓度范围内,可显著促进BMSCs的迁移,降低Ⅰ型胶原与Ⅲ型胶原RNA和蛋白相对表达量的比值,并可提高透明质酸的表达水平。

VEGF-C、SDF-1、S1P对慢性心力衰竭患者发生主要不良心血管事件的预测价值

目的 探讨血管内皮生长因子C(VEGF-C)、基质细胞衍生因子-1(SDF-1)、1-磷酸鞘氨醇(S1P)对慢性心力衰竭(CHF)患者发生主要不良心血管事件(MACE)的预测价值。方法 选取2021年1月至2023年1月湛江中心人民医院收治的149例CHF患者为研究对象。根据是否发生MACE将患者分为MACE组(54例)和对照组(95例)。比较两组VEGF-C、SDF-1、S1P的水平,采用二元logistic回归分析进行多因素分析,并以ROC曲线评价预测CHF患者发生MACE的价值。结果 MACE组的VEGF-C和S1P水平低于对照组,而SDF-1水平高于对照组,差异有统计学意义(P <0.05)。VEGF-C和S1P均是CHF患者发生MACE的独立保护因素,而SDF-1是CHF患者发生MACE的独立危险因素(P <0.05)。VEGF-C和S1P单独检测的1-AUC分别为0.806和0.811,而SDF-1单独检测的AUC为0.844,VEGF-C、SDF-1、S1P三者联合的AUC为0.938(P <0.01)。结论 VEGF-C、SDF-1、S1P均是CHF患者发生MACE的影响因素,并在MACE的预测中均有一定的价值,其中3种指标联合检测的预测效能最高。

血清IL-35、SDF-1在儿童腹型过敏性紫癜诊断中的临床意义

目的 探讨血清白细胞介素(IL)-35、基质细胞衍生因子-1(SDF-1)在儿童腹型过敏性紫癜(HSP)诊断中的临床意义。方法 选取2018年2月至2020年2月在该院治疗的160例早期HSP患儿作为研究对象,根据是否为腹型HSP分为腹型HSP组(80例)和其他类型HSP组(80例)。根据腹型HSP患儿早期病情程度分为轻度组26例、中度组26例和重度组28例。选取同期80例健康儿童作为对照组。采用酶联免疫吸附试验检测血清IL-35和SDF-1水平。采用Pearson相关分析腹型HSP患儿血清IL-35水平与SDF-1的相关性;采用多因素Logistic回归分析腹型HSP发生的影响因素;采用受试者工作特征(ROC)曲线分析血清IL-35和SDF-1对腹型HSP的诊断价值。结果 腹型HSP组血清IL-35水平明显低于其他类型HSP组、对照组(P<0.05),腹型HSP组血清SDF-1水平明显高于其他类型HSP组、对照组(P<0.05)。随着腹型HSP患儿病情程度的加重,血清IL-35水平明显降低(P<0.05),SDF-1水平明显升高(P<0.05)。腹型HSP患儿血清IL-35水平与SDF-1呈负相关(r=-0.594,P<0.05)。IL-35水平升高是发生腹型HSP的保护因素(P<0.05),SDF-1水平升高是发生腹型HSP的危险因素(P<0.05)。血清IL-35、SDF-1以及二者联合诊断腹型HSP的曲线下面积(AUC)分别为0.783、0.765、0.805,二者联合诊断的AUC优于血清IL-35、SDF-1单独诊断。结论 腹型HSP患儿血清IL-35水平降低,SDF-1水平升高,血清IL-35和SDF-1联合检测可更好地诊断腹型HSP。