Effect of Xiaoxuming Decoction on activation of astrocytes in acute cerebral ischemia/reperfusion injury / 中国中药杂志

This study investigated the effect of Xiaoxuming Decoction(XXMD) on the activation of astrocytes after cerebral ischemia/reperfusion(I/R) injury. The model of cerebral IR injury was established using the middle cerebral artery occlusion method. Fluorocitrate(FC), an inhibitor of astrocyte activation, was applied to inhibit astrocyte activation. Rats were randomly divided into a sham group, a model group, a XXMD group, a XXMD+FC group, and a XXMD+Vehicle group. Neurobehavioral changes at 24 hours after cerebral IR injury, cerebral infarction, histopathological changes observed through HE staining, submicroscopic structure of astrocytes observed through transmission electron microscopy, fluorescence intensity of glial fibrillary acidic protein(GFAP) and thrombospondin 1(TSP1) measured through immunofluorescence, and expression of GFAP and TSP1 in brain tissue measured through Western blot were evaluated in rats from each group. The experimental results showed that neurobehavioral scores and cerebral infarct area significantly increased in the model group. The XXMD group, the XXMD+FC group, and the XXMD+Vehicle group all alleviated neurobehavioral changes in rats. The pathological changes in the brain were evident in the model group, while the XXMD group, the XXMD+FC group, and the XXMD+Vehicle group exhibited milder cerebral IR injury in rats. The submicroscopic structure of astrocytes in the model group showed significant swelling, whereas the XXMD group, the XXMD+FC group, and XXMD+Vehicle group protected the submicroscopic structure of astrocytes. The fluorescence intensity and protein expression of GFAP and TSP1 increased in the model group compared with those in the sham group. However, the XXMD group, the XXMD+FC group, and XXMD+Vehicle group all down-regulated the expression of GFAP and TSP1. The combination of XXMD and FC showed a more pronounced effect. These results indicate that XXMD can improve cerebral IR injury, possibly by inhibiting astrocyte activation and down-regulating the expression of GFAP and TSP1.

Effect and mechanism of estrogen on EPCs function in diabetic rats / 药学实践杂志

Objective To explore the effect and mechanism of estrogen on endothelial progenitor cells（EPCs）function in diabetic rats. Methods EPCs were isolated from bone marrow of rats and characterized by fluorescence microscopy and flow cytometry. Rat diabetic model was established via streptozotocin induction. The bone marrow was taken to culture EPCs. EPCs of diabetes were incubated with Estrogen 10 nmol/L for 24h. The functions and proliferation of EPCs in vitro were detected. The levels of MnSOD and NO in EPCs and TSP-1 in supernatant were assayed. Results Compared with control group, EPCs proliferation, adhesion and angiogenesis functions were impaired in diabetic rats. The level of MnSOD and NO in diabetic EPCs were significantly decreased, while TSP-1 protein level in the supernatant increased. The above changes can be reversed with estrogen incubation. Conclusion Estrogen improved the EPCs migration and tubule formation in diabetic rats. The mechanism may be related to the reduction of oxidative stress and downregulation of TSP-1 expression in diabetic EPCs.

Effect of Modifide Wenjingtang on JAK2/STAT3 Signal Pathway and Downstream Factors Protein Expression in Endometriosis Rats / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect and mechanism of modified Wenjingtang on endometriosis （EM） rats with kidney deficiency and blood stasis. Method:The 10 from 105 SPF female healthy SD rats were randomly selected as the blank group. The rest constructed the rat model of kidney deficiency and blood stasis by compound factorial method. After the model was successfully established， 10 rats were randomly selected as the sham operation group， with only laparotomy and no intima suture， and the remaining rats were established with EM kidney deficiency and blood stasis type by autologous intimal transplantation. Fifty rats which were randomly selected from 56 successful rats were treated with the modified Wenjingtang （5，10，20 g·kg^-1） and danazol group（63 mg·kg^-1）， 1 time daily ， for 4 weeks. The endometrial tissues of each group were stained with hematoxylin eosin （HE） to observe the histopathology. The levels of inflammatory factors interleukin-10 （IL-10） and interleukin-17 （IL-17） in serum supernatant were detected by enzyme linked immunosorbent assay （ELISA）. Measuring the length（D₁），width （D₂） and height （D₃） of the heterotopic foci in each group before and after treatment. Then calculating the volume of them. The expression of tyrosine kinase 2（JAK2），transcription factor 3 （STAT3），phosphorylation transcription factor 3 （p-STAT3）， vascular endothelial growth factor （VEGF）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and thrombospondin-1 （TSP-1） were detected by immunohistochemistry （IHC）. The expression of VEGF，TNF-<italic>α</italic> and TSP-1 was detected by Western blot. Result:Microscopic pathological observation showed that the endometrial glandular cells of the blank group were arranged in order， and the glandular and stromal cells grew well， compared with the blank group， the endometrial structure of the model group was complete， showing a cavity like or annular closed structure， with cyst formation， and the epithelium was cubic or columnar epithelium， most of the epithelial cells had secretion， the stroma was dense， and the matrix showed a little fibrosis There were a few glands and inflammatory cell infiltration. Compared with the blank group， the content of IL-10 in serum of model group was significantly decreased （<italic>P</italic><0.01）， and the content of IL-17 was significantly increased （<italic>P</italic><0.01）， the protein expression of JAK2， STAT3，p-STAT3， VEGF， TNF-<italic>α</italic> in endometrial tissue of model group was significantly increased （<italic>P</italic><0.05）， and the expression of TSP-1 protein was significantly decreased （<italic>P</italic><0.05）. Compared with the model group， the serum IL-10 content of rats in modified Wenjingtang treatment group increased significantly （<italic>P</italic><0.01）， the IL-17 content decreased significantly <italic>（P</italic><0.01）， and the volume of ectopic foci decreased significantly （<italic>P</italic><0.01）. While the level of JAK2，STAT3，p-STAT3，TNF-<italic>α</italic>，VEGF protein in intimal tissue of modified Wenjingtang high and middle dose group decreased significantly （<italic>P</italic><0.05） and the level of TSP-1 protein increased significantly （<italic>P</italic><0.05）. Conclusion:Modified Wenjingtang can inhibit the invasion of ectopic foci in EM rats with kidney deficiency and blood stasis， the mechanism may be related to the intervention of immune barrier and block angiogenesis function mediated by JAK2/STAT3 signaling pathway activation.

Effect of TSP-1 gene on angiogenesis of osteosarcoma and its mechanism / 实用肿瘤学杂志

Objective The aim of this study was to investigate the effect of TSP-1 gene on angiogenesis in human osteosar-coma and its mechanism of action. Methods MG-63 cells were transfected with constructing pBPLV-shRNA-TSP-1 vector and pBPLV-TSP-1 expression vector. Cell viability was measured by CCK8,and its invasive ability was measured by Transwell assay. The expression of CD36 in intracells was detected by immunofluorescence. The expression levels of TSP-1,CD36,p38MAPK,VEGF, VEGFR-1,EGF and PDGF were detected in MG-63 cells by qRT-PCR and Western blot. Results The cell viability and inva-sion ability were significantly increased after transfected pBPLV-TSP-1 vector compared with the empty vector group(P<0. 05), and significantly decreased after transfected pBPLV-shRNA-TSP-1 vector( P<0. 05). The expression of TSP-1,EGF,P38, PDGF,VEGF and VEGFR -1 at mRNA and protein levels was significantly increased after transfection pBPLV -TSP -1 ( P <0. 05),and significantly decreased after transfection pBPLV-shRNA-TSP-1 vector(P<0. 05). Conclusion TSP-1 gene can promote the proliferation and invasion of MG-63 cells,and promote the formation of human osteosarcoma,indicating its mechanism related to the increase of growth factors EGF,VEGF,PDGF and activation of P38-MAPK pathway.

Analysis on application value of the detections of TSP-1 and VEGF in patients with lung cancer / 中国医学装备

Objective: To investigate the application value of the detections of thrombin sensitive protein-1(TSP-1) and vascular endothelial growth factor (VEGF) in patients with lung cancer. Methods: 90 patients with lung cancer were divided into lung cancer metastasis group (47 cases) and lung cancer non-metastasis group (43 cases) according to the pathological diagnosis results, and 40 persons whose results of physical examination were healthy were selected as the control group. The levels of TSP-1 and VEGF of the three groups were observed, and Spearman analysis was used to explore the correlations between TSP-1, VEGF and lung cancer, respectively. Results: The levels of TSP-1 and VEGF of platelet-poor plasma in the three groups were lower than the detection limit of the kit. And the levels of TSP-1 in platelet of the lung cancer metastasis group and non-metastatic group were significantly lower than those of the control group (t=-11.307, t=-6.295, t=12.851, t=9.732, P<0.05). In the platelet of lung cancer metastasis group, the level of TSP-1 was significantly lower than that of lung cancer non-metastasis group (t=-5.777, P<0.05), and the level of VEGF of metastasis group was significantly higher than that of lung cancer non-metastasis group (t=4.624, P<0.05). The results of Spearman correlation analysis showed that the correlation between platelet TSP-1 and lung cancer was negative correlation (r=-0.697, P<0.05), and that between platelet VEGF and lung cancer was positively correlation (r=0.757, P<0.05). Conclusion: The changes of TSP-1 and VEGF of platelet of patients with lung cancer are significantly, which are closely related to the progress of disease of patients with lung cancer. Therefore, the judgment about diagnosis and metastasis for patients with lung cancer have certain application value.

Effects of Hedysari Polysaccharide on Expressions of TSP-1 and PDGF-B in Retina of Diabetic Rats / 中国中医药信息杂志

Objective To observe the effects of Hedysari Polysaccharide (HPS) on the expressions of TSP-1 and PDGF-B in the retina of diabetic rats;To discuss the protective effect and possible mechanism on diabetic retinopathy. Methods The diabetic model was established by intraperitoneal injection of streptozotocin. 50 male SPF Wistar rats were randomly divided into 5 groups:model group, calcium dobesilate group, and HPS high-, medium-, and low-dose group, extra 10 rats were set as the normal group, 10 rats in each group. Each administration group was given relevant medicine for gavage, while model group and normal control group were given same amount NS for gavage, once a day for 8 weeks. The mRNA and protein expression of TSP-1 and PDGF-B were detected by qRT-PCR and immunohistochemistry. The retinal structure was observed by HE staining. Results HE staining showed that each layer of the retina of the model group was clear and complete, but the outer nucleus layer became looser, thinner and more disorderly, and the number of ganglion cells decreased slightly; the administration groups were improved markedly compared with the model group. Compared with the normal control group, the mRNA level and protein expression of retina TSP-1 on the model group dramatically dropped (P<0.01), and those of PDGF-B strikingly increased (P<0.01);Compared with the model group, the mRNA level and protein expression of retina TSP-1 on alladministration groups rose (P<0.05, P<0.01), and those of PDGF-B went down (P<0.01); Compared with all other administration groups, there was statistical significance in the mRNA level and protein expression of retina TSP-1 and PDGF-B on HPS high-dose group (P<0.05, P<0.01). Conclusion HPS may prevent the angiogenesis and proliferation in diabetic retinopathy process through adjusting the content of TSP-1 and PDGF-B in retina of diabetic rats so as to protect the retina.

Inhibition of Angiogenesis by the First Type I Repeat Peptides of Thrombospondin-1 / 체질인류학회지

Angiogenesis is the fundamental biological phenomenon in the development of vertebrates and various pathophysiological process such as cancer, inflammation and wound healing. Thrombospondin-1 is a well-known anti-angiogenic molecule which is distributed in the extracellular matrix of various tissues. The second and third type I repeats of human TSP-1 have inhibitory effects on endothelial cell migration and induce angiogenesis inhibition. However the role of the first type I repeat was not elucidated. In addition, the first type I repeat of bovine TSP-1 has CSVTCG amino acid sequence which is known to have anti-angiogenic activity. In the present study, we compared the inhibition of angiogenesis to investigate the role of the first type I repeat of the human and bovine TSP-1. Matrigel was mixed with or without TSR-1 peptides and then injected into C57BL/6J mice. We compared angiogenesis inhibition activity by hemoglobin assay, microvessel density and optical density value after 7 days. Furthermore, inhibition of angiogenesis was confirmed on CAM assay by TSR-1 peptides. For in vitro angiogenesis assay, TSR-1 peptides were treated on the proliferation, migration, and tube formation assay of HUVEC. Apoptosis effect of TSR-1 peptides was confirmed by apoptosis assay kit and flow cytometry. Bovine and human TSR-1 peptides blocked neovascularization in in vivo Matrigel plug assay and CAM assay at 10 microM. Bovine TSR-1 peptides have shown stronger angiogenesis inhibition in bFGF-induced angiogenesis than human TSR-1 and CSVTCG peptides. However, all of TSR-1 peptides inhibit migration and tube formation of HUVEC in in vitro. Furthermore, these peptides also induced apoptosis of HUVEC. These results suggest that TSR-1 peptides of bovine and human TSP-1 have angiogenesis inhibition activity.

Expression and significance of TSP-1 and BMP-7 in nasal mucosa of chronic rhinosinusitis / 基础医学与临床

Objective To explore the expression of thrombospondin-1 ( TSP-1 ) and bone morphogenetic protein-7 ( BMP-7 ) in mucus membrcane of chronic rhinosinusitis ( CRS ) and its potential significance in pathogenesis . Methods TSP-1 ,BMP-7 and TGF-β1 were detected by ELISA .Results The expression level of TSP-1 and TGF-β1 increased gradually with the advanced stage of CRSsNP ( P<0.05 ) .The expression of BMP-7 decreased gradu-ally with the advanced stage of CRSsNP ( P<0.05 ) .The expression of TSP-1 and TGF-β1 decreased gradually with the advanced stage of CRSwNP ( P<0.05 ) .The expression level of BMP-7 increased gradually with the ad-vanced stage of CRSwNP ( P<0.05 ) .Conclusions The expressions of TSP-1 and BMP-7 were abnormal in nasal mucosa of CRS , which might be involved in the pathogenesis of CRS .

Correlation analysis between serum thrombospondin-1 level and disease activity in patients with rheumatoid arthritis / 中华老年医学杂志

Objective To detect serum level of thrombospondin-1 (TSP-1) in rheumatoid arthritis (RA) patients,and analyze the correlation between serum TSP-1 level and disease activity in elderly versus young and middle aged RA patients in order to provide the basis for diagnosis and evaluation of RA.Methods Peripheral blood was collected from 98 RA patients (including 33 elderly,65 young and middle-aged RA patients) and 58 healthy controls.Serum TSP-1 levels were detected by enzyme linked immunosorbent assay (ELISA).Difference in TSP-1 level between RA patients and healthy controls was analyzed.Correlations of TSP-1 level with disease activity parameters of number of tender joints and swelling joints,disease activity score (DAS28 score) and erythrocyte sedimentation rate (ESR),levels of C-reactive protein (CRP),rheumatoid factor (RF),immunoglobin G (IgG),immunoglobin A (IgA) and immunoglobin M (IgM) were analyzed in elderly versus young and middle-aged RA patients.Spearman's and Pearson's correlation analysis were performed.Results TSP-1 level was significantly higher in RA patients than in healthy controls (P＜0.01).TSP-1 level was correlated with number of swelling joints and DAS28 score in RA patients (r=0.246 and 0.241,both P＜0.05).In elderly RA patients,TSP-1 level was correlated with number of swelling joints (r=0.377,P＜0.05),meanwhile significantly positive correlations of TSP-1 level with DAS28 score and rheumatoid factor level were observed in young and middle-aged RA patients (r =0.243 and 0.326,both P＜ 0.05).Conclusions TSP-1 may play roles in the development of RA and its determination may benefit evaluating disease activity.In elderly RA patients,TSP-1 level may reflect severity of rheumatoid arthritis and may be a novel biomarker to the evaluation of disease severity.

Expression of cyclooxygenase-2, urokinase plasminogen activator and thrombospondin-1 in gastric carcinoma and surrounding peripheral lymph nodes / 第二军医大学学报

Objective: To investigate the expression of cyclooxygenase-2 (COX-2), urokinase plasminogen activator(u-PA), and thrombospondin-1 (TSP-1) in gastric carcinoma and the surrounding lymph nodes, so as to discuss their roles in the invasion and metastasis of gastric carcinoma. Methods: The expression of COX-2, u-PA, and TSP-1 in the samples was examined by tissue microarray and S-P immunohistochemistry staining. Results: The positive rates of COX-2, u-PA, and TSP-1 in gastric carcinoma were 67.7%, 78.1%, and 78.6%, respectively, which were significantly higher than those in the surrounding lymph nodes (40.0%, 6.7%, and 40.0%, respectively, P0.05). COX-2, u-PA, and TSP-1 expression was not significantly different between metastatic lymph nodes and corresponding gastric carcinoma. COX-2 expression was positively correlated with u-PA, and TSP-1 expression. TSP-1 expression was not correlated with u-PA expression. Conclusion: COX-2, u-PA, and TSP-1 are highly expressed in the gastric carcinoma. COX-2 expression is correlated with depth of invasion. U-PA expression is significantly related with depth of invasion and lymph node metastasis.

Elevated Expression of p73 and Its Relation to VEGF and TSP-1 Production in Cervical Carcinoma / 대한산부인과학회지

OBJECTIVE: Measure the over-expression of p73 and analyze as the prognostic as well as angiogenic factor of cervical cancer by comparing the degree of expression of VEGF and TSP-1 by RT-PCR. METHODS: 7 normal and 37 cervical cancer specimens were put through RT-PCR and the expression of p73, VEGF and TSP-1 were measured. After immunohistochemical staining, the number of microvessels was counted. With the level of expression, investigated the relationship with the clinicopathological characteristics and the number of microvessels. RESULTS: 57% of cancer tissues showed abnormally high levels of p73 mRNA. In quantitative genomic DNA PCR, the p73 was over-expressed in the transcription level. Through allotyping with Sty I polymorphism, the over-expression of p73 was due to the transcription activity of the silent allele. In RT-PCR-SSCP analysis of over-expressed specimens, sequence alterations was not seen. In 73%, VEGF was over-expressed while TSP-1 was under-expressed in 35%. There was no association between the number of microvessels with the over-expression of p73 and VEGF, but inversely associated with the under-expression of TSP-1. There was no correlation between the over-expression of p73 and the clinicopathological characteristics. The over-expression of p73 coincided 80% with the over-expression of VEGF, and 40% with the under-expression of TSP-1. CONCLUSION: These data support the expression of p73 was increased in cervical cancer tissues and was associated with the over-expression of the VEGF but not associated with the under-expression of TSP-1. The biological and clinical significance of the over-expression of p73 should be studied further in the future.

Effect of Recombinant N-segment of Thrombospondin-1 on the Proliferation of ECV304 / 中国生物工程杂志

Objective: To construct an expression system containing the N-terminal segment of Thrombospondin-1 (TSP-1) in E. coli. To investigate the activity of TSF. Methods: thbs1 gene fragment was amplified from human fetal cord vein endothelial cells and inserted into plasmid pET32c ( + ) which was then transfected and expressed in E. coli. Investigate the effect of recombinant TSF to the proliferation of ECV304 in vitro with MTT. The expression level of CD36 was assayed by FACS. Results: Recombinant TSF was purified. TSF could restrain the proliferation of ECV304 with CD36 low-expression. Conclusions:Low dose of rTSF was a latent assistant treatment of anti-cancer.

Expression of vascular endothlial growth factor and throbospondin-1 in bladder cancer / 中国康复理论与实践

@#ObjectiveTo observe the expression of vascular endothlial growth factor(VEGF) and throbospondin-1(TSP-1) in different grade bladder cancer.MethodsSpecimens of 70 cases of bladder transitional cell cancer including Grade Ⅰ 28 cases,Grade Ⅱ 27 cases,Grade Ⅲ 15 cases(according to WHO pathological grade) were stained with immunohistochemistry method.The expressing values of VEGF and TSP-1 were calculated by mean vascular density(MVD).All data was analyzed with SPSS 10.0,and with expression of VEGF and TSP-1 was detected with immunohistochemisrty double staining.Finally,the result of expression was observed by laser scan with focus microscope.ResultsThere was a positive correlation between the cancer grade and the expression of VEGF,and the expression of Grade Ⅰ was significant difference with that of Grade Ⅱ(P<0.01),but there was no difference between Grade Ⅱ and Grade Ⅲ(P>0.1).Expression of TSP-1 decreased with grade increasing but not showing a negative correlation,while,Grade Ⅰ was significant difference with that of Grade Ⅱ and Grade Ⅲ(P<0.01),Grade Ⅱ was not differcnt with Grade Ⅲ(P>0.1).In 28 Grade Ⅰ cases,values of MVD of VEGF and TSP-1 had no correlation(rs)=0.167,P>0.1).Double immunostaining detecting with laser scan with focus microcope indicate showed that VEGF and TSP-1 appeared in same bladder cancer specimen.ConclusionExpression of VEGF is gradually increased from Grade Ⅰ to Grade Ⅲ,and VEGF is a promoting factor of angiogenesic bladder cancer.Expression of TSP-1 is the strongest in Grade Ⅰ cases,and TSP-1 is an inhibiting angiogenesic factor,which inhibiting function only in early stage.Immunofluorescent staining can only provide evidence of together expression of VEGF and TSP-1,cannot evaluate the degree of expression.

Weakening of the repressive YY-1 site on the thrombospondin-1 promoter via c-Jun/YY-1 interaction

Thrombospondin-1 (TSP-1) level is tightly regulated at the transcriptional level. To determine the detailed molecular mechanisms of TSP-1 expression, nine serial 5'-deletion constructs of the human genomic tsp-1 promoter (nucleotides -2,220 to +756) were prepared, inserted into luciferase reporter plasmids, and transiently transfected into the Hep3B human hepatocarcinoma cell. Among the nine 5'-deletion constructs, pTSP-Luc-4 (-767~+756) had consistently decreased luciferase activity with or without PMA stimulation, whereas a further truncated construct [pTSP-Luc-4' (-407~+756)] had increased levels of expression. By searching the nucleotides from -767 to -407, a consensus binding sequence (5'-CCATTTT-3') for the repressor Yin Yang-1 (YY-1) at nucleotide -440 was identified. The suppression induced by this site was weakened in the presence of the region upstream of nucleotide -767 (pTSP-Luc-1 and -2). Nuclear protein directly bound to an oligonucleotide containing the repressive YY-1 sequence but the binding capacity of the sequence was decreased by the increased c-Jun levels. Moreover, proteins immunoprecipitated with anti-YY-1 revealed an interaction between c-Jun and YY-1 factor. These data suggest that the repressive YY-1 site of the tsp-1 promoter could not be functional via activating positive cis-elements on the upstream from this site and weakened via c-Jun/YY-1 interactions.

Cell-type specific regulation of thrombospondin-1 expression and its promoter activity by regulatory agents

Thrombospondin-1 (TSP-1), a multifunctional protein that is able to function as a negative regulator of solid tumor progression and angiogenesis, is normally present at a very low level but rapidly elevated in pathological tissues. To understand the cellular regulation of TSP-1 expression, the mode of it's expression in Hep3B, SK-HEP-1, and porcine aortic endothelial (PAE) cells was examined in the presence of all-trans retinoic acid (ATRA), interleukin-6 (IL-6), interferon-gamma (IFN-gamma), or phorbol 12-myristate 13-acetate (PMA). ATRA or IL-6 induced a dose-dependent increase of TSP-1 protein and mRNA levels in PAE cells, while they negatively regulated TSP-1 expression in the Hep3B and SK-HEP-1 cells. In contrast, PMA showed just the opposite effects on the TSP-1 expression in the same cells. IFN-gamma had little effect on TSP-1 level in Hep3B and PAE cells. The TSP-1 expression in SK-HEP-1 cells by these agents showed a close resemblance to that of liver cells rather than that of the endothelial cell line. Possible TSP-1 promoter-mediated responses by ATRA, IL-6, IFN-gamma, or PMA in Hep3B and PAE cells examined with luciferase activity of TSP-LUC reporter plasmid showed that levels of TSP-1 promoter activity were lower than that of the expressed TSP-1 protein and mRNA levels. Transfection of c-Jun and/or RARalpha expression vectors into Hep3B and PAE cells resulted in the enhanced TSP-1 promoter activity as well as the increments of of its protein and mRNA level. These results suggest that regulatory agents-induced TSP-1 expression may be attributed to mRNA stability and/or translational activation in concert with transcriptional activation and TSP-1 expression may be independently controlled via each signal pathway stimulated by PMA or ATRA.

The Expression of Vascular Endothelial Growth Factor and Thrombospondin-1 in Wilms' Tumor / 대한비뇨기과학회지

PURPOSE: With the process of neoangiogenesis being linked to the growth and metastasis of various tumors, anticancer therapeutics with a basis in the suppression of neoangiogenesis has recently been receiving attention. In this study, we tried to clarify the immunoreactivities of vascular endothelial growth factor (VEGF), major angiogenic inducer and thrombospondin-1 (TSP-1), major angiogenic inhibitor in human Wilms' tumor and its clinicopathological significance. MATERAILS AND METHODS: Utilizing immunohistochemical staining, we assessed the immunoreactivities of VEGF and TSP-1 in archival tissues of 29 Wilms' tumors and 25 normal kidneys. Also, we assessed the relationship between expression of each factor and clinicopathological parameters in 29 cases of Wilms' tumors. RESULTS: Immunoreactivities of VEGF and TSP-1 were detected mainly in the cytoplasm of the tubular cells in normal kidneys. In Wilms' tumors, whereas VEGF was detected in the cytoplasm of the tumor cells and peritumoral stromal tissues, but TSP-1 only in the peritumoral stromal tissues. Immunohistochemical expression patterns of each factor were divided into two groups according to the area of immunoreactivity (negative: OR =10%). VEGF immunoreactivity was detected in 25 (100%) normal kidneys and in 20 (69%) Wilms' tumors. However, TSP-1 immunoreactivity was detected in 24 (97%) normal kidneys and in 3 (10%) Wilms' tumors. Therefore, although no significant difference was observed between the expressions of VEGF and TSP-1 in normal kidney, the TSP-1 immunoreactivity was significantly lower than VEGF immunoreactivity in Wilms' tumors. A relatively higher rate of positive expression of TSP-1 was observed in the patients with no demonstrable lymph node metastasis. Also, as for the VEGF, maximal diameter of the tumor was larger in the positive expression group. However, it proved otherwise for TSP-1 as the negative expression group demonstrated tumors with larger maximal diameters. CONCLUSIONS: Our study demonstrated that the TSP-1 immunoreactivity was significantly lower than VEGF immunoreactivity in Wilms' tumors, and disease progression has a tendency to be found in the VEGF-positive cases and TSP-1 negative cases. We suggest that the growth and metastasis of Wilms' tumor may be influenced mainly by TSP-1 decrease rather than VEGF increase.