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在SDF-1介导内皮细胞参与血管形成中的作用

目的探讨CXCR7在SDF-1诱导内皮细胞参与血管生成中的作用。方法用Western blot和流式细胞仪检测脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)中CXCR7和CXCR4的表达;利用小分子拮抗剂分别阻断CXCR4或CXCR7,然后通过MTT、Transwell小室法及三维基质胶血管样结构形成实验分别考察CXCR7和CXCR4对SDF-1诱导HUVECs增殖、迁移及管样结构形成能力的影响。结果 HUVECs细胞中同时高表达CXCR4和CXCR7;CXCR7的拮抗剂显著抑制SDF-1诱导HUVECs的增殖(P<0.01);而SDF-1诱导HUVECs迁移却被CXCR4的拮抗剂阻止(P<0.01);CXCR7和CXCR4的拮抗剂均显著阻止SDF-1诱导HUVECs形成血管样结构(P<0.01)。结论 CXCR7和CXCR4在SDF-1诱导HUVECs参与血管生成的过程中均起着不可或缺却又不尽一致的作用,SDF-1诱导HUVECs的增殖主要通过CXCR7发挥作用,而CXCR4主要参与SDF-1诱导HUVECs的迁移,两者共同参与SDF-1诱导HUVECs形成管样结构的调节。

SDF-1/CXCR4促进骨髓基质细胞迁移的作用

目的研究骨髓基质细胞的CXCR4表达,SDF-1对BMSCs迁移、趋化作用,分析促进迁移规律和机理。为改善和提高骨髓基质细胞迁移能力寻找更有效的途径。为临床骨髓基质细胞更广泛的应用奠定基础。方法体外培养大鼠骨髓基质细胞,应用免疫组化方法对骨髓基质细胞的CXCR4受体表达水平进行检测,运用微孔隔离室穿越实验方法,研究SDF-1对骨髓基质细胞迁移、趋化作用,并用SDF-1阻断剂加以证实。结果CXCR4在各代骨髓基质细胞中表达小于5%,SDF-1可促进骨髓基质细胞迁移。结论虽然CXCR4在骨髓基质细胞中表达量很少,但是对促进骨髓基质细胞迁移起到至关重要的作用。

AMD3100抑制胰腺癌细胞增殖和侵袭的体外实验

目的探讨CXCR4特异性受体阻滞剂AMD3100对胰腺癌细胞增殖和侵袭能力的影响及其可能的作用机制。方法用CCK-8检测经不同浓度AMD3100处理后人胰腺癌细胞株SW1990的增殖。Matrigel胶铺设的transwell小室检测经AMD3100处理后SW1990的侵袭能力。RT-PCR法检测AMD3100处理后SW1990细胞血管内皮生长因子(VEGF)及基质金属蛋白酶-9(MMP-9)的表达。结果 AMD3100能够抑制SW1990细胞的增殖,尤其是对基质细胞源性因子1(SDF-1)诱导的增殖抑制作用最为明显。AMD3100亦能够减弱SW1990细胞的迁移及侵袭能力。AMD3100处理SW1990细胞后,其VEGF及MMP-9的表达下降。结论 CXCR4特异性受体阻断剂AMD3100通过下调VEGF及MMP-9的表达抑制胰腺癌细胞的增殖,减弱胰腺癌细胞的侵袭及转移能力。

VEGF、SDF-1、Ki-67、PCNA及Survivin与翼状胬肉的关系分析

目的:分析血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质细胞衍生因子-1(stromal cellderived factor 1,SDF-1)、肿瘤增殖抗原(Ki-67)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及生存素(Survivin)与翼状胬肉的关系。方法:选取2013-01/2015-05于本院进行诊治的79例106眼翼状胬肉患者为观察组,同时期的79例正常结膜者为对照组,然后将两组的组织VEGF、SDF-1、Ki-67、PCNA及Survivin阳性细胞数分级及染色强度分级进行比较,并比较其中不同性别、分期及分型患者的检测结果,同时以Logistic分析上述指标与翼状胬肉的关系。结果:观察组的组织VEGF、SDF-1、Ki-67、PCNA及Survivin阳性细胞数分级及染色强度分级均高于对照组,不同分期及分型患者的检测结果也存在一定差异(均P<0.05),而不同性别患者间的检测结果则无明显差异(均P>0.05),经Logistic分析显示,上述组织指标均与翼状胬肉有密切的关系。结论:翼状胬肉组织中的VEGF、SDF-1、Ki-67、PCNA及Survivin表达呈现异常状态,上述指标均与翼状胬肉有密切的关系。

SDF-1/CXCR4生物轴活化诱导对鼠骨髓源性内皮祖细胞生物学性能的影响

目的观察基质细胞衍生因子-1(SDF-1)及其特异性受体CXCR4生物轴对体外培养的鼠骨髓源性内皮祖细胞(EPCs)生物学性能的影响。方法以密度梯度离心法获取大鼠骨髓单个核细胞,由Dil-ac-LDL和FITC-UEA-1荧光双染鉴定后确认为EPCs。传代培养后,加入不同浓度SDF-1溶液,培养48 h,然后分别采用MTT法检测EPCs的增殖、Transwell实验检测细胞迁移、细胞爬片行免疫组化检测EPCs的CXCR4表达。结果 SDF-1能明显促进EPCs的增殖,并呈显著的剂量依赖性(P<0.01);同时,SDF-1能显著改善EPCs的迁移能力(P<0.01或0.05),并能增强EPCs的CXCR4表达(P<0.01或0.05)。结论 SDF-1/CXCR4轴对鼠骨髓源性EPCs的数量和功能有重要的促进作用。

超声联合SDF-1微泡对糖尿病性心肌病大鼠心肌损伤的保护作用

目的研究超声联合基质细胞衍生因子-1(SDF-1)微泡对糖尿病性心肌病大鼠心肌损伤的保护作用。方法将40只SD大鼠随机分为空白组、模型组、SDF-1组、SDF-1+超声组、微泡+超声组,每组8只。除空白组外,其余组均采用高脂饲料喂养和腹腔注射1%链脲佐菌素(STZ)方法制备糖尿病性心肌病模型。造模成功后,SDF-1组经尾静脉输入SDF-1溶液10 ng/kg;SDF-1+超声组经尾静脉输入SDF-1溶液10ng/kg,然后采用2%戊巴比妥钠1 mL/kg腹腔注射麻醉大鼠,利用超声治疗仪辐照大鼠心前区10 min;微泡+超声组经尾静脉输入含有10 ng/kg SDF-1的微泡混合液,超声照射同SDF-1+超声组。干预4 d后,留取血清标本用于检测肌酸激酶(CK)、乳酸脱氢酶(LDH)、心肌肌钙蛋白I(cT nI)水平;干预8周后,超声心动图检测各组大鼠左心室舒张末期内径(LVIDd)、左心室舒张末压(LVEDP)和左心室射血分数(LVEF),Western blot法检测各组心肌组织中CXC族趋化因子受体4(CXCR4)蛋白表达情况,实时定量PCR法检测各组心肌组织中转化生长因子β1(TGF-β1)mR NA表达情况。结果干预4 d后,各组血清CK、LDH和cT nI水平比较差异均无统计学意义(P均>0.05)。干预8周后,各干预组LVIDd、LVEDP均明显低于模型组(P均<0.05),LVEF均明显高于模型组(P均<0.05),微泡+超声组各指标改善情况均明显优于SDF-1组和SDF-1+超声组(P均<0.05);模型组及各干预组CXCR4蛋白表达水平和TGF-β1mR NA表达水平均明显高于空白组(P均<0.05),各干预组TGF-β1mR NA表达水平均明显低于模型组(P均<0.05);微泡+超声组CXCR4蛋白表达水平明显高于其他组(P均<0.05),TGF-β1mR NA表达水平均明显低于SDF-1组和SDF-1+超声组(P均<0.05)。结论超声联合SDF-1微泡可延缓糖尿病性心肌病大鼠心肌纤维化。

SDF-1/CXCR4轴在肺癌中的表达及意义

肺癌的转移和复发是造成肺癌患者死亡的主要原因,肿瘤细胞的趋化迁移能力日益引起人们的关注,目前对于肿瘤趋化迁移能力的研究多趋向于肿瘤细胞趋化因子方面。CXCR4是目前研究最多的趋化因子,其导致肺癌转移的传导通路和调节机制还有待进一步阐明,合理设计以它们为靶点的药物,将为肺癌的治疗开创广阔的前景。

SDF-1/HGF融合蛋白介导的大鼠间充质干细胞对造血干细胞增殖和趋化作用的影响

目的：研究基质细胞衍生因子-1（stromal cell-derived factor-1,SDF-1）/肝细胞生长因子（hepatocyte growth factor,HGF）融合蛋白介导的骨髓间充质干细胞（bone marrow derived stroma cell,BMSCs）对造血干细胞（hematopoietic stem cell,HSC）增殖和趋化能力的影响,以期望通过联合移植的方式重建肝组织。方法：培养鉴定3~4周龄SD大鼠BMSCs及大鼠HSCs。荧光显微镜下观测重组腺病毒p AD-SDF-1-（Gly Ser）3-HGF-IRES-GFP感染BMSCs 1、3、7、14 d后的感染效率,同时ELISA检测SDF-1和HGF的表达。制备BMSCs细胞滋养层,HSCs种植于有滋养层的24孔板中,分为3组：HSCs加培养基组（A1组）;HSCs加BMSCs组（B1组）;HSCs加SDF/HGF融合蛋白介导的BMSCs组（C1组）,检测共培养1、3、7、14 d后HSCs的增殖倍数。利用transwell培养体系,按实验要求分为3组：HSCs加细胞培养液组（A2组）;HSCs加BMSCs组（B2组）;HSCs加SDF/HGF融合蛋白介导的BMSCs（C2组）,测定HSCs的迁移指数。结果：重组腺病毒p AD-SDF-1-（Gly Ser）3-HGF-IRES-GFP感染BMSCs 1、3、7、14 d的感染效率分别为（25.28±3.72）%、（82.22±3.58）%、（64.85±4.48）%、（28.35±2.73）%,感染后第3天的感染效率最高（与第1、7、14天相比较,F=718.05,P=0.000;F=66.80,P=0.000;F=642.78,P=0.000）。重组腺病毒p AD-SDF-1-（Gly Ser）3-HGFIRES-GFP感染BMSCs 1、3、7、14 d后,SDF-1的表达分别为：（1.02±0.15）μg/ml,（4.51±0.52）μg/ml,（2.18±0.19）μg/ml,（1.22±0.21）μg/ml;HGF的表达分别为：（1.38±0.32）ug/ml,（5.01±0.43）μg/ml,（3.10±0.26）μg/ml,（1.63±0.27）μg/ml。感染后第3天SDF-1与第1、7、14天比较和HGF与第1、7、14天比较的表达明显增高（P均=0.000）。BMSCs和HSCs共培养第3、7、14天,B1组、C1组中HSCs的扩增倍数明显高于A1组（P均=0.000）。同时C1组中HSCs的扩增倍数明显高于B1组（P均=0.000）。transwell显示B2、C2组的迁移指数明显高于A2组,同时C2组的迁移指数明显高于B2组。结论：SDF-1/HGF融合蛋白介导的BMSCs可能在促进HSCs的增殖及趋化方面起到重要作用。

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.

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.

基质细胞衍生因子-1基因转染对人胃癌细胞株SGC7901增殖、耐药的影响及其机制

目的观察基质细胞衍生因子-1(SDF-1)基因转染对人胃癌细胞株SGC7901增殖、耐药的影响,并探讨其可能作用机制。方法 取对数生长期 SGC7901 细胞分为实验组、阴性对照组、空白对照组,实验组、阴性对照组分别转染pcDNA3.1-SDF-1质粒、pcDNA3.1质粒(空白质粒),空白对照组不做任何处理。质粒转染48 h时分别采用RT-PCR法、Western bloting法检测细胞SDF-1 mRNA及蛋白,质粒转染24、48、72、96 h时MTT法测算三组细胞增殖能力(以光密度OD值表示),质粒转染48 h时观察三组细胞对奥沙利铂的耐药性(以奥沙利铂对细胞的半数抑制浓度IC 50 表示),质粒转染48 h时流式细胞仪测算三组细胞凋亡率,质粒转染48 h时Western blotting法检测三组细胞上皮型钙黏蛋白( E-cadherin)、神经型钙黏蛋白(N-cadherin)及波形蛋白(Vimentin)、微管相关蛋白1轻链3Ⅱ(LC3-Ⅱ)自噬相关因子(Beclin 1)。结果 与空白对照组、阴性对照组比较,培养48 h时实验组细胞SDF-1 mRNA及蛋白相对表达量均升高( P 均<0.05)。与空白对照组、阴性对照组比较,培养24、48、72、96 h实验组细胞OD值均升高( P 均<0.05)。质粒转染48 h时奥沙利铂对实验组、空白对照组、阴性对照组细胞的IC 50 值分别为 28.039 ±3.011、10.403±1.253、10.612±1.044(μg/mL),与空白对照组、阴性对照组比较,奥沙利铂对实验组细胞的IC 50 值升高( P 均<0.05)。质粒转染48 h时实验组、空白对照组、阴性对照组细胞凋亡率分别为39.10%± 2.26%、53.90%±3.19%、51.30%±3.37%,与空白对照组、阴性对照组比较,实验组细胞凋亡率降低( P 均< 0.05 )。质粒转染48 h时倒置显微镜下可见实验组细胞存在上皮-间质转化(EMT)现象,加入奥沙利铂后实验组细胞自噬活动增加;与空白对照组、阴性对照组比较,实验组细胞E-cadhein蛋白相对表达量降低,N-cadhein、Vimentin、LC3-Ⅱ、Beclin 1蛋白相对表达量均升高( P 均<0.05)。结论 SDF-1基因过表达可促进SGC7901细胞增殖,显著提高SGC7901细胞对奥沙利铂的耐药性,可能与SDF-1促进EMT、诱导细胞自噬增加进而抑制细胞凋亡有关。

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.

基质细胞衍生因子受体在造血干/祖细胞宫内移植中的作用

目的探讨基质细胞衍生因子受体(CXCR4)在造血干/祖细胞(HSC)宫内移植归巢中的作用。方法分离人脐血CD34+细胞,用流式细胞仪检测SCF、IL-6处理前后细胞表面CXCR4(CD184)的表达及在Transwell板中的迁移率。在BALB/c胎鼠孕13～14d期间,经胎鼠腹腔注射经不同处理的CD34+细胞,胎鼠出生1个月后取骨髓,用流式细胞仪检测人CD45细胞。结果预处理后表达CD184的CD34+细胞的百分数由原来的9.58%±1.56%上升为19.32%±3.64%。CD34+/CXCR4high细胞迁移率显著增高,但迁移作用可以被antiCXCR4mAb和PTX明显抑制。SCF和IL-6预处理组胎鼠人CD45细胞阳性率显著高于其他组。抗CXCR4抗体或PTX预处理组人CD45细胞检出率显著降低。结论增加CD34+细胞CXCR4的表达有助于宫内移植HSC的归巢,HSC宫内移植归巢过程依赖CXCR4受体,SDF-1/CXCR4调节信号通过Gi蛋白进行跨膜传导。

超声联合一氧化氮微泡对大鼠心肌组织的生物学效应

目的:本研究旨在探讨超声联合一氧化氮(NO)微泡的作用对大鼠心肌组织局部基质衍生因子-1(SDF-1)的影响,及其对心肌组织作用的安全性。方法:将32只SPF级SD雄性大鼠随机分为4组,每组8只。第1、2、3组分别经尾静脉输入1 ml的NO微泡、普通脂质微泡以及PBS,并采用频率为1 MHz、强度为1 W/cm2的超声辐照大鼠心前区,辐照时间为10 min;第4组为空白对照组。4 h后每组处死4只大鼠,留取心肌组织进行HE染色,观察局部的炎症反应情况;同时留取血清标本用于检测肌酸激酶(CK),乳酸脱氢酶(LDH)和肌钙蛋白I(TnI)的含量。1周后处死剩余大鼠,取心肌组织进行RT-PCR和Western blot分别检测SDF-1的基因及蛋白相对表达量。结果:辐照4 h后留取心肌组织行HE染色显示各组均可见少量炎性细胞浸润,组间没有明显差异;血清学指标检测显示,第1组与第2组血清CK和LDH高于第3组和第4组,且差异有统计学意义(P<0.05);第1组血清TnI低于第2组,但高于第3组和第4组,且差异有统计学意义(P<0.05);辐照1周后留取的心肌组织行RT-PCR和Western blot分析结果均显示第1组SDF-1相对表达量最多,各组间进行两两比较,差异均有统计学意义(P<0.05)。结论:超声联合NO微泡辐照大鼠心肌组织,与内含全氟显气体的普通脂质微泡一样,对心肌组织无明显破坏;但能使大鼠心肌组织SDF-1表达增加,其程度高于应用超声联合普通脂质微泡,因此,NO微泡应用于干细胞归巢将优于目前普通脂质微泡。

氧诱导视网膜病模型中曲安奈德抑制新生血管生长的机制

目的观察氧诱导视网膜病模型(OIR)中曲安奈德(TA)对CD14+细胞聚集及VEGF表达的干预作用,初步探讨曲安奈德抑制视网膜新生血管生长的可能机制。方法清洁级C57BL/6哺乳期小鼠36只(共36个眼球),随机将其分为四组:①正常对照组(6个眼球),6只17日龄正常小鼠先行FFA眼底造影,然后每鼠随机摘取一个眼球,行眼球石蜡切片HE染色。②单纯高氧组(6个眼球),6只17日龄OIR模型小鼠处理同单纯对照组。③TA高氧组(12个眼球),12只12日龄OIR模型小鼠随机一眼注射TA 2μL,再于17日龄后行FFA眼底造影后摘除术眼,其中6个眼球行眼球石蜡切片HE染色及视网膜CD14和VEGF免疫组化染色,另6个眼球行视网膜VEGF mRNA的real-time PCR检测。④BSS高氧(12个眼球),12只12日龄OIR模型小鼠随机一眼注射BSS 2μL,余处理同TA高氧组。用t检验两两比较各组视网膜突破内界膜的内皮细胞核数,视网膜CD14与VEGF免疫组化染色的平均吸光度值(IOD/AOI),及VEGF mRNA的相对含量值(2-ΔCt×105)。结果与正常对照组相比,高氧诱导组突破视网膜内界膜的内皮细胞核数目明显增多(t=29.62,P<0.001)。与BSS高氧组相比,TA高氧组突破视网膜内界膜的内皮细胞核数明显减少(t=19.879,P<0.001)。TA高氧组和BSS高氧组小鼠视网膜神经上皮均可见VEGF,CD14阳性染色,但BSS高氧组的VEGF、CD14含量明显高于TA高氧组(t=-2.743,P<0.05;t=-3.592,P<0.01)。并且视网膜SDF-1与CD14的蛋白含量存在正相关(r=0.662,n=12,P<0.05)。视网膜VEGF mRNA表达在BSS高氧组也明显高于TA高氧组(t=-4.754,P<0.005)。结论 TA能有效抑制小鼠视网膜新生血管形成,其机制可能是通过阻遏循环中CD14+细胞的聚集,进而减少VEGF等细胞因子表达而发挥作用。

穿山龙总皂苷对白介素-1β诱导的成纤维样滑膜细胞基质细胞衍生因子-1及IκB激酶表达的影响

目的观察穿山龙总皂苷(Rhizoma Dioscoreae nipponicae,RDN)对体外白介素-1β(r IL-1β)诱导的成纤维样滑膜细胞(FLS)基质细胞衍生因子-1(SDF1)及IκB激酶(IKK)表达的影响。方法原代培养FLS,取第3代对数生长期的细胞将其分为正常组、模型组及给药组,模型组给予10μg/L r IL-1β诱导FLS增殖,给药组给予10μg/L r IL-1β及100μg/L穿山龙总皂苷共同孵育,正常组不予处理,孵育72 h后采用Western blot法测定各组大鼠FLS中SDF1及IKK(p-IKK)的表达水平。结果与正常组比较,模型组SDF1及p-IKK表达水平明显升高(P<0.01);与模型组比较,给药组FLS中SDF1及p-IKK表达水平明显降低(P<0.01)。结论穿山龙总皂苷可抑制SDF-1及IκB激酶的激活,可能通过对SDF-1的表达调控进而调控IKK的表达。

三种不同免疫状态小鼠皮肤创面愈合过程中基质细胞衍生因子-1的表达

目的观察3种不同免疫状态小鼠皮肤创面愈合过程中基质细胞衍生因子-1（SDF-1）的基因表达。方法采用差异性贴壁法分离纯化BALB／C雄性小鼠骨髓源性干细胞（BMSCs），取第3代BMSCs进行成脂、成骨诱导鉴定；同时另取第3代BMSCs，经尾静脉注射到3．5Gy放射性损伤后的雌性BALB／C鼠、裸鼠、SCID鼠体内，小鼠背部皮肤制作一直径为1．5cm的创面，建立骨髓移植嵌合体皮肤创伤动物模型，观察创面的愈合过程，并于伤后1、3、5、7、14d获取创面组织，应用半定量逆转录-聚合酶链反应（RT—PCR）检测创面组织中SDF。1的基因表达。结果提取的BM—SCs可以成功进行成骨、成脂诱导分化；创面愈合过程中BALB／C鼠SDF-1基因表达于伤后1d即有增高，5d达峰值（P〈0．01），然后逐渐下降，14d创面基本愈合时接近对照组。裸鼠、SCID鼠创面中SDF-1基因表达于伤后逐渐增高，7d达峰值（P〈0．01），与BALB／C鼠比较，SDF-1基因表达峰值延后，14d创面未愈，SDF-1基因表达仍高于对照组。结论皮肤创面愈合过程中，SDF-1趋化BMSCs，参与创面愈合的炎症反应期和增殖期的修复过程，在创面愈合中起着重要作用；机体的免疫功能也将影响BMSCs募集，从而影响创面愈合。