Thrombospondin peptide ABT-898 inhibits inflammation and angiogenesis in a colitis model

AIM: To evaluate the efficacy of the improvedthrombospondin mimetic peptide ABT-898 in a murine model of ulcerative colitis. METHODS: The dextran sodium sulfate(DSS) was used for the induction of colitis in both TSP-1 deficient(TSP-1-/-) and wild type(WT) mice during 7 d. While mice were receiving the DSS dissolved in the drinking water, the ABT-898 peptide was dissolved in sterile 5% glucose solution and delivered using mini pumps subcutaneously implanted. Plasma samples were analyzed for interleukin(IL)-6 by ELISA assay and colonic tissues were harvested, fixed and processed for histological evaluation. Immunohistochemistry using antibodies for the detection of CD31 and MECA in endothelial cells was performed. Inflammation was graded in colonic sections and the number of microvessels in each lesion was assessed. Activation of signal transducer and activator of transcription 3(STAT3) in colonic samples was quantified by immunohistochemistry and Western blotting using antibodies against total STAT3 and phosphorylated STAT3(p STAT3)(Ser727).RESULTS: Treatment with ABT-898 considerably diminished the inflammatory response in WT and TSP-1-/- mice(P < 0.0001 in both groups vs control). Identification of blood vessels highlighted by CD31/MECA immunohistochemistry, showed significantly reduced vessel counts in colitic lesions of WT and TSP-1-/- mice treated with ABT898(TSP-1-/- controls/TSP-1-/- treated, P = 0.0002; WT controls/WT treated, P = 0.0005). Consistently, IL-6 was significantly diminished in plasma samples of TSP-1-/- and WT treated with the peptide when compared to the control mice(P = 0.0002 and P = 0.0148, respectively). p STAT3 positive cells were quantified in WT and TSP-1-/- treated with ABT-898. A significant decrease in positive cells for p STAT3 was observed in treated mice(TSP-1-/- controls/TSP-1-/- treated, P = 0.0089; WT/WT treated, P = 0.0110). These results were confirmedby Western blotting analyses showing lower levels of p STAT3 in colitic lesions from mice treated with the peptide ABT-898.CONCLUSION: These findings indicate that the new peptide ABT-898 ameliorates inflammation and angiogenesis and might be a therapeutic alternative in IBD and inflammatory diseases.

Anti-osteoarthritis effect of a combination treatment with human adipose tissue-derived mesenchymal stem cells and thrombospondin 2 in rabbits

BACKGROUND Osteoarthritis(OA),a chronic age-related disease characterized by the slowly progressive destruction of articular cartilage,is one of the leading causes of disability.As a new strategy for treatment of OA,mesenchymal stem cells(MSCs)have the potential for articular cartilage regeneration.Meanwhile,thrombospondin 2(TSP2)promotes the chondrogenic differentiation of MSCs.AIM To investigate whether TSP2 induces chondrogenic differentiation of human adipose-derived MSCs(hADMSCs)and potentiates the therapeutic effects of hADMSCs in OA rabbits.METHODS We investigated the chondrogenic potential of TSP2 in hADMSCs by analyzing the expression of chondrogenic markers as well as NOTCH signaling genes in normal and TSP2 small interfering RNA(siRNA)-treated stem cells.Anterior cruciate ligament transection surgery was performed in male New Zealand white rabbits,and 8 wk later,hADMSCs(1.7×10^6 or 1.7×10^7 cells)were injected into the injured knees alone or in combination with intra-articular injection of TSP2(100 ng/knee)at 2-d intervals.OA progression was monitored by gross,radiological,and histological examinations.RESULTS In hADMSC culture,treatment with TSP2 increased the expression of chondrogenic markers(SOX9 and collagen Ⅱ)as well as NOTCH signaling genes(JAGGED1 and NOTCH3),which were inhibited by TSP2 siRNA treatment.In vivo,OA rabbits treated with hADMSCs or TSP2 alone exhibited lower degree of cartilage degeneration,osteophyte formation,and extracellular matrix loss 8 wk after cell transplantation.Notably,such cartilage damage was further alleviated by the combination of hADMSCs and TSP2.In addition,synovial inflammatory cytokines,especially tumor-necrosis factor-α,markedly decreased following the combination treatment.CONCLUSION The results indicate that TSP2 enhances chondrogenic differentiation of hADMSCs via JAGGED1/NOTCH3 signaling,and that combination therapy with hADMSCs and TSP2 exerts synergistic effects in the cartilage regeneration of OA joints.

Thrombospondins and synaptogenesis

Here, we review research on the mechanisms underlying the ability of thrombospondin to promote synaptogenesis and examine its role in central nervous system diseases and drug actions. Thrombospondin secreted by glial cells plays a critical role in synaptogenesis and maintains synapse stability. Thrombospondin regulates synaptogenesis through receptor a26-1 and neuroligin 1, and promotes the proliferation and differentiation of neural progenitor cells. It also participates in synaptic remodeling following injury and in the action of some nervous system drugs.

In vivo expression of thrombospondin-1 suppresses the formation of peritoneal adhesion in rats

BACKGROUND Formation of intraperitoneal adhesions is one of the major complications after abdominal surgery, which may lead to bowel obstruction. Thrombospondin 1(TSP-1) is an extracellular matrix modulating glycoprotein during tissue regeneration and collagen deposition.AIM To evaluated the therapeutic potential of overexpressed TSP-1 in suppressing pelvic adhesion formations in rat models.METHODS Pelvic adhesion was induced in anesthetized rats by laparotomy cecal abrasion.The animals were randomly assigned to treatment of local application with Seprafilm(an antiadhesive bioresorbable membrane) or adenoviral vectors encoding mouse TSP-1(AdTSP-1) on the surfaces of the injured cecum. The severity of the peritoneal adhesions was evaluated by blinded observers 14 d later.RESULTS Compared with control(no treatment) group, the application of Sperafilm significantly reduced the formation of adhesion band, and local administration of AdTSP-1 on the injured cecum the also attenuated the severity of peritoneal adhesion score. However, systemic delivery of AdTSP-1 did not affect the formation of adhesion.CONCLUSION We conclude that therapeutic approaches in inducing regional overexpression of TSP-1 may serve as alternative treatment strategies for preventing postoperative peritoneal adhesion.

Thrombospondin 1 promotes synaptic formation in bone marrow-derived neuron-like cells

In this study, a combination of growth factors was used to induce bone marrow mesenchymal stem cells differentiation into neuron-like cells, in a broader attempt to observe the role of thrombospondin 1 in synapse formation. Results showed that there was no significant difference in the differentiation rate of neuron-like cells between bone marrow mesenchymal stem cells with thrombospondin induction and those without. However, the cell shape was more complex and the neurites were dendritic, with unipolar, bipolar or multipolar morphologies, after induction with thrombospondin 1. The induced cells were similar in morphology to normal neurites. Immunohistochemical staining showed that the number of positive cells for postsynaptic density protein 95 and synaptophysin 1 protein was significantly increased after induction with thrombospondin 1. These findings indicate that thrombospondin 1 promotes synapse formation in neuron-like cells that are differentiated from bone marrow mesenchymal stem cells.

The Role of Angiopoietin-1 and Thrombospondin-1 in the Kidney of Rats Subject to 5/6 Nephrectomy

The expression of angiopoietin- 1 （Ang- 1） and thrombospondin- 1 （TSP- 1） in 5/6 subtotal nephrectomy （STN） rats model, and its correlation to the renal microvasculature injury were investigated. Rat 5/6 STN model was established in adult male SD rats, and the sham-operated group and 5/6 STN group were set up. The renal function and histopathological changes were examined at the 1st, 2nd, 4th, 8th and 12th week after operation. The expression orAng-1, TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry. From 2nd to 8th week after operation, Ang-1 was significantly expressed in glomeruli of rats with STN. Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation, and subsequently decreased after the 12th week. The expression of TSP-1 was increased significantly in STN group. As compared with sham-operated group, the CD31 expression was significantly down-regulated from the 2nd week. The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points. The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd, 4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points, while decreased evidently at the 12th week as compared with that in sham-operated group. It is concluded that there are changes in the mRNA expression of Ang-1, and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN, which may be involved in the remnant renal microvasculature injury.

Circulating thrombospondin-2 in patients with moderate-to-severe chronic heart failure due to coronary artery disease

Chronic heart failure（CHF）remains a leading cause of morbidity and mortality.In the current study,we aimed to evaluate the predictive value of circulating fhrombospondin-2（TSP-2）for cumulative survival in patients with ischemic CHF due to coronary artery disease（CAD）.The results showed that during a median follow-up of2.18 years,21 participants died and 106 subjects were hospitalized repeatedly.The median circulating levels of TSP-2 in patients who survived and those who died were 0.63 ng/mL（95%CI=0.55-0.64 ng/mL）and 1.03 ng/mL（95%CI=0.97-1.07 ng/mL）（P〈0.001）.Circulating TSP-2 independently predicted all-cause mortality（OR=1.27;95%CI=1.08-1.59;P=0.002）,CHF-related death（OR=1.16;95%CI=1.02-1.50;P〈0.001）,and also CHF-related rehospitalization（OR=1.12;95%CI=1.07-1.25;P〈0.001）.In conclusion,among CAD patients with symptomatic CHF,increased circulating TSP-2 is correlated with increased 3-year CHF-related death,all-cause mortality,and risk for recurrent hospitalization.

Redistribution of Platelet Membrane Glycoprotein IV and Release of Intracellular α-granule Thrombospondin in Patients with Chronic Myelogenous Leukemia

The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular Q-granule thrombospondin (TSP) were examined and the inhibition of 5-thromboglobulin (&TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080±17010 molecules/platelet as compared with 13190±4810 from the controls (P<0,01), No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/III.(P>0. 05). The GPIV redistribution on active platelet membrane induced thrombin (1U/ml) from CML and healthy donors was 44320132310 and 228001 12700 molecules/platelet respectively (P<0. 01 ). The difference in the release of intracellular Q-granule TSP between CML and the control group was not found (P>0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high leveIs of β-TG and PF4 in sera inhibited release of intracellular a-granule TSP in vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of interna1 TSP pools is hindered by either β-TG or PF4 in sera.

Thrombospondin 1 Promotes Endoplasmic Reticulum Stress and Apoptosis in HK-2 Cells by Upregulating ATF6-CHOP

Objective The goal of this study is to investigate the role and mechanism of endoplasmic reticulum stress and apoptosis regulated by thrombospondin 1(TSP1)in human renal tubular epithelial cells(HK-2 cells).Methods HK-2 cells were exposed to high concentrations of glucose(HG).The endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA)was administered by transfecting TSP1 or an empty vector to explore the mechanism of the endoplasmic reticulum response regulated by TSP1 and stress in renal cell apoptosis.The effects of TSP1 and 4-PBA on the proliferation and apoptosis of HK-2 cells under HG conditions were assessed using Cell counting kit-8 and flow cytometry.Western blotting was used to detect the apoptosis-and endoplasmic reticulum stress-related protein expression regulated by TSP1 and 4-PBA.Results HG treatment induced high cell apoptosis,abundantly expressed TSP1 level and restrained viability in HK-2 cells.Overexpression of TSP1 significantly inhibited the proliferation of and facilitated apoptosis of HK-2 cells under HG conditions.Administration of endoplasmic reticulum stress inhibitor 4-PBA after overexpression of TSP1 antagonized the inhibitory proliferation and promoted apoptosis rate in HG-triggered HK-2 cells induced by TSP1 overexpression.4-PBA treatment significantly hindered the expression of endoplasmic reticulum stress markers,such as PERK,ATF4,ATF6,p-eIF2α,IRE1,CHOP and XBP1,suggesting that the administration of 4-PBA was successful.Conclusion Overexpression of TSP1 activated endoplasmic reticulum stress by regulating the ATF6-CHOP axis.TSP1 restrained cell proliferation,and promoted apoptosis and endoplasmic reticulum stress by activating the ATF6-CHOP axis.

No association between thrombospondin-4 A387P polymorphism and acute coronary syndrome in Chinese Han population

Objective： The thrombospondin-4 （TSP-4） gene G29926C （A387P） polymorphism was recently reported to be associated with an increased risk of MI （myocardial infarction） in American population. However, several subsequent studies produced controversial findings. The aim of this study was to explore the possible association between TSP-4 A387P polymorphism and ACS （acute coronary syndrome） in Chinese Han population. Methods：A case-control study including 412 patients with ACS and 337 controls free from CAD （coronary artery disease） was conducted. TSP-4 A387P polymorphism was determined by PCR （polymerase chain reaction） and RFLP （restriction fragment length polymorphism） analysis. Results：Slightly decreased frequency of GC genotype was observed in patients with ACS, compared with controls（5.3% vs. 7.1%）, but the difference did not reach statistical significance （P = 0.31 ）. Similarly, the prevalence of C allele was 2.7% and 3.6% for ACS and control groups, respectively （P = 0.32）. None of homozygote was detected for C allele. Further analyses in subjects subgrouped according to sex and age also showed no association of TSP-4 A387P polymorphism with ACS. Furthermore, after adjustment for conventional risk factors by multiple logistic regression analysis, the carrier prevalence of C allele did not differ significantly between ACS and control groups （OR = 0.85; 95% CI：0.45-1.59; P = 0.60）. Conclusion：The present study suggested that the TSP-4 A387P variant showed a low prevalence compared with western populations and failed to associate with an altered risk of ACS in Chinese Han population. The findings further supplement experimental data for TSP-4 gene study of coronary disease.

Thrombospondin-1 Expression in RPE and Choroidal Neovascular Membranes

Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Tissue sections from normal human fetal and adult eyes and surgically removed CNVMs were immunostained for TSP-1 localization. Polymerase chain reaction and Western blotting were used to analyze TSP-1 mRNA and protein from human RPE cells, respectively. TSP-1 in the supernatant of cultured RPE cells and eye explants were measured using enzyme-linked immunosorbent assay. MTT assay was used to evaluate the RPE survival after TSP-1 treatment. Results: The strongest immunostaining for TSP-1 was observed in the RPE monolayer around drusen in early AMD. The intensity of TSP-1 staining in normal eye sections was much weaker than that of early AMD and CNVM. TSP-1 mRNA was positive in cultured fetal and adult RPE cells. There was increasing secretion of TSP-1 into the supernatant of cultured RPE and eye explants. The specific band of TSP-1 was identified by Western blot. No significant inhibition of RPE survival was found with the exposure to TSP-1. Conclusions: TSP-1 expression in drusen and CNVM was upregulated and associated with RPE monolayer. TSP-1 may be a natural negative regulator for choroidal neovascularization.

Thrombospondin-1 Levels in Patients with Coronary Heart Disease

Background: Thrombospondin-1 (TSP1) is associated with atherosclerosis in animals with diabetes mellitus (DM), but the precise role of TSP-1 in human atherosclerosis remains unknown. Objectives: To investigate serum thrombospondin1 level in patients with coronary artery disease with and without type 2 DM and its relationship to coronary artery scoring systems. Methods: The study comprised 180 patients recruited from those underwent coronary angiography for suspected coronary artery disease (CAD) was approved by Institutional Review Board and Institutional Ethical Committee for Human Research of menoufia university hospital. They were divided according to presence of CAD and type 2 DM into 4 groups: Group I (n = 44 patients): Non diabetic subjects without CAD, Group II (n = 40 patients): Diabetic patients without CAD, Group III (n = 49 patients): Non diabetic patients with CAD and Group IV (n = 47 patients): Diabetic patients with CAD. Serum level of TSP-1 was measured in all groups and coronary artery scoring analysis was done. Results: Serum TSP-1 levels were higher in patients with CAD and DM than in other groups (P < 0.01) while the levels of serum TSP-1 in diabetic patients without CAD and non-diabetic patients with CAD didn’t show significant difference. Plasma TSP-1 levels were higher in patients with DM than those in patients without DM (582.95 ± 55.70 ng/mL vs. 516.91 ± 64.56 ng/ml, P <0.001). Plasma levels of TSP-1 were higher in patients with CAD than those in patients without CAD (569.54 ± 68.16 ng/mL vs. 525.17 ± 61.77 ng/ml, P < 0.001). Patients with CAD and DM (group IV) had significantly higher severity score and vessel score than those with CAD only (group III) (9.13 ± 3.40, 2.49 ± 0.69 vs. 7.37 ± 3.16, 2.02 ± 0.80 ng/ml, P < 0.05). Patients with three vessel disease had the highest serum TSP-1 level (581.32 ± 61.30 ng/ml) when compared to patients with one or two vessel disease. The highest diagnostic performance of serum TSP-1 level in prediction of coronary artery disease was more pronounced in presence of DM while the least diagnostic performance of serum TSP-1 was detected in absence of DM. Univariate logistic regression analysis of different variables for prediction of CAD showed that TSP-1 level was one of the independent predictors of CAD (OR 1.016, CI 1.010 - 1.023, P < 0.001). Conclusion: Serum TSP-1 level is higher in patients with CAD with and without type 2 DM and its level is an independent predictor of CAD, but the diagnostic performance of serum TSP-1 level in prediction of CAD is more pronounced in presence of type 2 DM.

Thrombospondin-1 expression correlates with angiogenesis in experimental cirrhosis

AIM:To investigate the significance of Thrombospondin-1 (TSP-1) expression and its relationship with angiogenesis during experimental fibrosis. METHODS:Cirrhosis was induced in male Wistar rats by intraperitoneal administration of diethyl nitrosamine (DEN). The serial sections from liver tissues were stained with anti-CD34 and anti-TSP-1 antibodies before being quantitated by light microscopy. RESULTS:Our results showed that of TSP-1 expression gradually increases according to the severity of fibrosis (GroupⅠvs group Ⅱ, Group Ⅲ and Group Ⅳ;Group Ⅱ vs group Ⅲ and group Ⅳ;group Ⅲ vs group Ⅳ, P < 0.05). Moreover, TSP-1 expression was found to be correlated with angiogenesis (P < 0.05). CONCLUSION:The correlative evidence of the link between TSP-1 and fibrosis or angiogenesis provided by this study suggests that besides its role as a strong promoter of transforming growth factor-β1 (TGF-β1), TSP-1 might have an additional role in liver fibrogenesis by stimulating angiogenesis and this protein could be a potential target to prevent fibrogenesis in chronic inflammatory diseases of the liver.

Expression of Thrombospondin-1 is Correlated with Microvessel Density in Prostate Cancer

OBJECTIVE To observe the expression of thrombospondin-1 ( TSP-1) in prostate cancer, and examine its expression in relation to angiogenesis. METHODS The expression of TSP-1 and microvessel density (MVD) were studied in 22 prostate cancer patients by using immunohistochemistry. RESULTS Positive expression of the TSP-1 protein was detected in 16 (72.7%)of the 22 cases. Most of the positive staining for TSP-1 was seen in the cytoplasm of the cancer cells, but some was in the extracellular matrix. The mean MVD in the 22 prostate cancer cases was 71.21±31.14 vessels per 100 high field of vision. Tumors with an elevated expression of TSP-1 showed a high MVD resulting in a correlation between TSP-1 immunopositivity and microvessel density that was highly significant (r= 0.54, P=0.009). CONCLUSION TSP-1 is strongly expressed in most prostate cancers and is associated with neovascularization. Therefore TSP-1 is a likely contributor to the extensive neovascularization in prostate cancer and increased TSP-1 expression might participate in an angiogenic phenotype.

敲除TSP-1对小鼠急性肝损伤保护作用的机制

目的研究敲除血小板反应蛋白1(thrombospondin 1,TSP-1)在四氯化碳(CCl_(4))所致小鼠急性肝损伤中的保护作用。方法选取C57BL/6J小鼠16只,TSP-1基因敲除小鼠8只:C57BL/6J小鼠分为WT组和WT+CCl_(4)组,TSP-1基因敲除小鼠为TSP-1^(-/-)+CCl_(4)组;WT+CCl_(4)和TSP-1^(-/-)+CCl_(4)组给予0.5 mL CCl_(4)+9.5 mL/kg橄榄油进行腹腔注射建立急性肝损伤模型,WT组注射橄榄油10 mL/kg作为对照。通过HE染色以及血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)观察各组小鼠肝脏损伤情况及炎症反应程度;同时检测肝组织丙二醛(MDA)和谷胱甘肽(GSH)的含量并通过免疫组化和TUNEL检测凋亡的变化。结果敲除TSP-1对CCl4诱导的急性肝损伤具有明显的改善作用,表现为血清AST、ALT、IL-6和TNF-α的降低和肝细胞坏死的减少,以及降低MDA含量和升高GSH含量,减少肝组织中凋亡及caspase-3的表达。结论敲除TSP-1对CCl_(4)诱导的小鼠急性肝损伤具有明显的保护作用,其机制可能与抑制炎症、氧化应激和细胞凋亡相关。

Correlation between the expression of thrombospondin-1 and neovascularization in mucoepidermoid carcinoma

Background Researchers have recently demonstrated that thrombospondin-1 （TSP-1） has an important function in regulating neovascularization. Whether it inhibits or accelerates neovascularization, however, is still controversial. We found few reports about the correlation between TSP-1 and vascularization in mucoepidermoid carcinoma. In this research, the distribution and expression of TSP-1 in mucoepidermoid carcinoma were investigated. We also analyzed （1） the correlation between the expression of TSP-1 and microvessel density （MVD）, as an indicator of neovascularization activity, and （2） the effect of TSP-1 on neovascularization and tumor growth in the subcutaneous xenotransplanted model of mucoepidermoid carcinoma. Method （1） The sites and intensity of expression of TSP-1 and the MVD were analyzed in 45 cases of mucoepidermoid carcinoma after surgery by the method of streptavidin-peroxidase （SP） immunohistochemistry; and （2） recombinant human thrombospondin-1 （rhTSP-1） was injected twice a week for five consecutive weeks around the tumor in the subcutaneous xenotransplanted tumor model of mucoepidermoid carcinoma in nude mice. Each week, the tumor size was measured, in order to draw the growth curve of the xenotransplanted tumor model of mucoepidermoid carcinoma, and MVD was measured. Results （1） The positive expression of TSP-1 protein was 57.78% （26/45）. Most positive staining for TSP-1 was found in the cytoplasm of the cancer cells, while some staining occurred in the extracellular matrix. The mean MVD in 45 cases of m ucoepidermoid carcinoma was 58.17±19.77 per 100 visual fields. Tumors with a high expression of TSP-1 showed a low MVD value, and the TSP-1 immunocompetence and microvessel density showed a significant negative correlation （rs=-0.947, P 〈0.001）. （2） The xenotransplanted tumors with the injection doses of 1.25, 0.75 and 0.25 ug/ml respectively were 36.97%, 53.36% and 73.61% of the size of the control group （（451±92）, （651±113）, （898±86） and （1220±157） mm^3 respectively, F=-53.167, P 〈0.001）, and their weights were respectively 35.14%, 51.35% and 70.27% of the control group （（1.3±0.5）, （1.9±0.5）, （2.6±0.3）, and （3.7±0.7） g respectively, F=-62.669, P 〈0.001）. Their MVDs were 25.00%, 45.93%, and 72.20% respectively of the control group and concentration dependent （15.43±3.45, 28.35±4.24, 44.57±3.35 and 61.73±5.43 per 100 visual fields respectively, F=54.582, P 〈0.001）. Conclusions The TSP-1 has a higher expression in mucoepidermoid carcinoma and the expression has a significant negative correlation with neovascularization. The TSP-1 inhibits neovascularization and tumor growth, and it might be a new biological therapy for treatment of patients with mucoepidermoid carcinoma.

不同Child-Pugh分级肝硬化患者血清TSP-1、球蛋白/胆碱酯酶的表达水平差异及其疾病预后危险因素分析

目的分析不同Child-Pugh分级肝硬化患者血清凝血酶敏感蛋白-1(TSP-1)、球蛋白/胆碱酯酶的表达水平差异及其疾病预后危险因素。方法回顾性选取2020年2月至2023年2月新疆医科大学第一附属医院收治的70例肝硬化患者作为主要研究对象,根据Child-Pugh分级将其分为Child-Pugh A级组(n=20),Child-Pugh B级组(n=34),Child-Pugh C级组(n=16),另选取同期在本院进行体检的50名健康人群作为对照组。采用酶联免疫吸附试验法检测4组及肝硬化不同预后患者的血清TSP-1、球蛋白、胆碱酯酶、球蛋白/胆碱酯酶表达水平;采用双变量Spearman相关性检验血清TSP-1、球蛋白、胆碱酯酶、球蛋白/胆碱酯酶与肝硬化患者Child-Pugh分级和预后的相关性;建立多因素Logistic模型分析影响肝硬化患者预后的独立危险因素,并绘制受试者工作特征(ROC)曲线分析血清TSP-1、球蛋白/胆碱酯酶对肝硬化预后的预测价值。结果与对照组比较,Child-Pugh A级组、Child-Pugh B级组、Child-Pugh C级组患者的血清TSP-1、球蛋白、球蛋白/胆碱酯酶表达水平较高,血清胆碱酯酶表达水平较低;与Child-Pugh A级组患者比较,Child-Pugh B级组、Child-Pugh C级组患者的血清TSP-1、球蛋白、球蛋白/胆碱酯酶表达水平较高,血清胆碱酯酶表达水平较低;与Child-Pugh B级组比较,Child-Pugh C级组患者的血清TSP-1、球蛋白、球蛋白/胆碱酯酶表达水平较高,血清胆碱酯酶表达水平较低,差异均有统计学意义(P<0.05)。与预后良好组比较,预后不良组血清TSP-1、球蛋白、球蛋白/胆碱酯酶表达水平较高,血清胆碱酯酶表达水平较低,差异均有统计学意义(P<0.05)。肝硬化患者血清TSP-1、球蛋白/胆碱酯酶与Child-Pugh分级和预后均呈正相关(P<0.05)。多因素Logistic分析结果显示,Child-Pugh分级、TSP-1、球蛋白/胆碱酯酶均是影响肝硬化患者预后的独立危险因素(P<0.05)。血清TSP-1、球蛋白/胆碱酯酶与TSP-1+球蛋白/胆碱酯酶预测肝硬化患者预后的曲线下面积值分别为0.814、0.824、0.885。结论血清TSP-1、球蛋白/胆碱酯酶异常表达与肝硬化Child-Pugh分级及其预后均存在一定关联,可作为肝硬化患者的Child-Pugh分级及预后的辅助预测指标。

特发性血小板减少性紫癜患儿血清LXA4、ADAMTS-1水平及其在预后评估中的价值

目的 探讨特发性血小板减少性紫癜(ITP)患儿血清脂氧素A4(LXA4)及含凝血酶敏感素1型基序的解聚素样金属蛋白酶(ADAMTS-1)水平以及二者在ITP预后评估中的价值。方法 将2020年8月至2022年8月于该院确诊为ITP的112例患儿纳入研究作为ITP组。另选取同期于该院体检的106例健康志愿者作为对照组。采用酶联免疫吸附法检测受检者血清LXA4、ADAMTS-1水平。采用Pearson相关分析患儿血清LXA4、ADAMTS-1水平的相关性。对不同病情及预后患儿的血清LXA4与ADAMTS-1水平进行比较。采用受试者工作特征(ROC)曲线分析血清LXA4与ADAMTS-1对ITP预后不良的预测价值。采用Logistic回归分析影响ITP患儿预后的因素。结果 与对照组相比,ITP组血清LXA4与ADAMTS-1的水平均升高(t=16.921、17.360,P<0.05)。Pearson相关分析显示,血清LXA4与ADAMTS-1呈正相关性(r=0.577,P<0.05)。与轻度组相比,中度组与重度组血清LXA4与ADAMTS-1表达水平较高(P<0.05);与中度组比较,重度组血清LXA4与ADAMTS-1水平较高(P<0.05)。预后不良组血清LXA4与ADAMTS-1水平均高于预后良好组(t=7.903、11.480,P<0.05);血清LXA4、ADAMTS-1单独及联合检测用于ITP预后不良预测的曲线下面积(AUC)分别为0.695、0.816、0.882,二者联合预测ITP预后的AUC大于LXA4单独预测的AUC(Z=3.363,P<0.05)和ADAMTS-1单独预测的AUC(Z=2.534,P<0.05)。Logistic回归分析显示,LXA4与ADAMTS-1是ITP预后不良的危险因素(P<0.05)。结论 ITP患儿血清LXA4与ADAMTS-1水平升高,联合检测LXA4、ADAMTS-1水平有助于评估患儿预后。

孕晚期血清血小板反应蛋白-1、D-二聚体及金属蛋白酶组织抑制物-1水平对瘢痕子宫再次妊娠患者产后出血的预测价值

目的探讨孕晚期血清血小板反应蛋白-1(THBS-1)、D-二聚体(D-D)及金属蛋白酶组织抑制物-1(TIMP-1)水平对瘢痕子宫再次妊娠患者产后出血(PPH)的预测价值。方法选择2020年6月至2022年8月新乡医学院第一附属医院收治的108例瘢痕子宫再次妊娠孕妇为研究对象,根据孕妇分娩后是否发生PPH分为PPH组(n=21)和非PPH组(n=87)。采集2组孕妇入院当天肘静脉血5 mL,应用酶联免疫吸附法检测2组孕妇血清THBS-1、D-D、TIMP-1水平。比较2组孕妇的基本临床资料及血清THBS-1、D-D、TIMP-1水平。采用多因素logistic回归分析瘢痕子宫再次妊娠孕妇发生PPH的影响因素,受试者操作特征(ROC)曲线分析血清THBS-1、D-D、TIMP-1水平对瘢痕子宫再次妊娠孕妇发生PPH的预测价值。结果PPH组人工流产次数≥2次、胎盘早剥、子宫切口撕裂、宫缩乏力、瘢痕厚度<0.3 cm占比及孕晚期血清THBS-1、D-D水平显著高于非PPH组,血清TIMP-1水平显著低于非PPH组(P<0.05)。宫缩乏力、D-D和THBS-1水平升高是瘢痕子宫再次妊娠孕妇PPH的独立危险因素(P<0.05),TIMP-1水平降低是瘢痕子宫再次妊娠孕妇PPH的保护因素(P<0.05)。血清THBS-1、D-D、TIMP-1联合预测瘢痕子宫再次妊娠孕妇PPH的曲线下面积大于三者单独预测(P<0.05)。结论孕晚期血清THBS-1、D-D、TIMP-1水平均可作为预测瘢痕子宫再次妊娠孕妇发生PPH的参考指标,且三者联合对瘢痕子宫再次妊娠孕妇发生PPH的预测效能更高。

Thrombospondin I activates the macrophage Toll-like receptor 4 pathway

Previously, we demonstrated that macrophages from thrombospondin 1 （TSP1）-deficient mice have a reduced inflammatory phenotype, suggesting that TSP1 plays a role in macrophage activation. In this study, we determined how TSP1 regulates macrophage function. We found that recombinant or purified piatelet human TSP1 treatment stimulated tumor-necrosis factor （TNF）-α expression in bone marrow-derived macrophages in a time- and dose-dependent manner. Toll-like receptor 4 （TLR4） expression （at the mRNA and protein levels） and nuclear factor-kappaB （NF-KB） activity were also stimulated by TSP1 treatment. The TSPl-mediated increase in TNF-a production was abolished in TLR4-deficient macrophages, suggesting that TSP1 activates macrophages through a TLR4-dependent pathway. TSP1 also stimulated TLR4 activation in macrophages in vivo. Furthermore, TSPl-mediated macrophage activation was attenuated by using a peptide or an antibody to block the association between TSP1 and CD36. Taken together, these data suggest that the stimulation of the macrophage TLR4 pathway by TSP1 is partially mediated by the interaction of TSP1 with its receptor, CD36.