scholarly journals Matrix metalloproteinase 9 Contributes to Glomerulosclerosis by Causing Profibrotic Changes in Podocytes and Glomerular Endothelial Cells

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Qiao X ◽  
◽  
Guo J ◽  
Chen J ◽  
Loron MC ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Confocal Microscopy ◽  
Interstitial Fibrosis ◽  
Physiological Saline ◽  
Wild Type ◽  
Glomerular Endothelial Cells ◽  
Adriamycin Nephropathy ◽  
Metalloproteinase 9 ◽  
Sirius Red

Background: Glomerulosclerosis is characterized by progressive (myo) fibroblast accumulation and collagen deposition involving profibrotic changes of podocytes and endothelial cells. A profibrotic role of MMP-9 in interstitial fibrosis has been reported. Whether MMP-9 plays a role in glomerulosclerosis is not clear yet. Methods: Mouse glomerulosclerosis model [Adriamycin Nephropathy (AN) model] was induced by a single adriamycin injection (10.2mg/kg, with physiological saline for controls) through tail vein in MMP-9-/- and wild-type control mice of BALB/c background. All animals were sacrificed at 4 weeks after injection. Albuminuria (albumin to creatinine ratio) and calculated GFR were measured. Gomori Trichrome (GT) and Sirius Red (SR) staining were used for assessment of glomerular fibrosis. Profibrotic changes of podocytes or glomerular endothelial cells were examined by confocal microscopy using immunofluorescence staining (IF) of desmin or a-SMA with P-cadherin or VEcadherin. Results: Albuminuria was reduced while GFR was increased in MMP-9-/- AN mice compared with those of wild-type mice. Confocal microscopy showed a significant decrease in podocytes double-stained with P-cadherin and desmin, demonstrating that MMP9-/- AN mice were protected from profibrotic changes in podocytes and glomerular endothelial cells. Glomerulosclerosis was significantly reduced in MMP9-/- AN mice compared to that of WT, as demonstrated by GT and SR staining. Conclusions: MMP-9 contributes to glomerulosclerosis at least in part by causing profibrotic changes in podocytes and glomerular endothelial cells.

MMP-9 contributes to glomerulosclerosis by causing profibrotic changes in podocytes and glomerular endothelial cells

2019 ◽  
Author(s):  
Xi Qiao ◽  
Jianhong Guo ◽  
M Loron ◽  
Lixin Liu ◽  
Padmashree Rao ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Confocal Microscopy ◽  
Interstitial Fibrosis ◽  
Wild Type ◽  
Glomerular Endothelial Cell ◽  
Glomerular Endothelial Cells ◽  
Adriamycin Nephropathy ◽  
Sirius Red ◽  
Wild Type Control

Abstract Background Glomerulosclerosis is characterized by progressive (myo)fibroblast accumulation and collagen deposition involving profibrotic changes of podocytes and endothelial cells. A profibrotic role of MMP-9 in interstitial fibrosis has been reported. Whether MMP-9 plays a role in glomerulosclerosis is not clear yet. Methods A mouse glomerulosclerosis model [adriamycin nephropathy (AN)] was produced in MMP-9-/- and wild-type control BALB/c mice. All animals were sacrificed at 4 weeks after injection. Albuminuria (albumin to creatinine ratio) and calculated GFR were measured. Gomori Trichrome (GT) and Sirius Red (SR) staining were used for assessment of glomerular fibrosis. Profibrotic changes of podocytes or glomerular endothelial cells were examined by confocal microscopy using immunofluorescence staining (IF) of desmin or α-SMA with P-cadherin or VE-cadherin. Results Albuminuria was reduced while GFR was increased in MMP-9-/- AN mice compared with those of wild-type mice. Confocal microscopy showed a significant decrease in podocytes double-stained with P-cadherin and desmin and decrease in glomerular endothelial cell co-staining with VE-cadherin and α-SMA, demonstrating that MMP9-/- AN mice were protected from profibrotic changes in podocytes and glomerular endothelial cells. Glomerulosclerosis was significantly reduced in MMP-9-/- AN mice compared to that of WT, as demonstrated by GT and SR staining. Conclusions MMP-9 contributes to profibrotic changes in podocytes and glomerular endothelial cells and thereby glomerulosclerosis.

Abstract 110: Thrombospondin Type-1 Domain-Containing Protein 1 (THSD1), an Intracranial Aneurysm Susceptibility Gene, Mediates Cell Adhesion in Human Umbilical Vein Endothelial Cells

Stroke ◽  
2015 ◽  
Vol 46 (suppl_1) ◽  
Author(s):  
Xiaoqian Fang ◽  
Dong H Kim ◽  
Teresa Santiago-Sim
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Focal Adhesion ◽  
Umbilical Vein ◽  
Adhesion Formation ◽  
Wild Type ◽  
Focal Adhesion Formation ◽  
Umbilical Vein Endothelial Cells

Introduction: An intracranial aneurysm (IA) is a weak spot in cerebral blood vessel wall that can lead to its abnormal bulging. Previously, we reported that mutations in THSD1 , encoding thrombospondin type-1 domain-containing protein 1, are associated with IA in a subset of patients. THSD1 is a transmembrane molecule with a thrombospondin type-1 repeat (TSR). Proteins with TSR domain have been implicated in a variety of processes including regulation of matrix organization, cell adhesion and migration. We have shown that in mouse brain Thsd1 is expressed in endothelial cells. Hypothesis: THSD1 plays an important role in maintaining the integrity of the endothelium by promoting adhesion of endothelial cells to the underlying basement membrane. Methods: Human umbilical vein endothelial cells are used to investigate the role of THSD1 in vitro . THSD1 expression was knocked-down by RNA interference. Cell adhesion assay was done on collagen I-coated plates and focal adhesion formation was visualized using immunofluorescence by paxillin and phosphorylated focal adhesion kinase (pFAK) staining. THSD1 re-expression is accomplished by transfection with a pCR3.1-THSD1-encoding plasmid. Results: Knockdown of THSD1 caused striking change in cell morphology and size. Compared to control siRNA-treated cells that exhibited typical cobblestone morphology, THSD1 knockdown cells were narrow and elongated, and were significantly smaller ( p <0.01). Cell adherence to collagen I-coated plates was also attenuated in THSD1 knockdown cells ( p <0.01). Consistent with this finding is the observation that the number and size of focal adhesions, based on paxillin and pFAK staining, were significantly reduced after THSD1 knockdown ( p <0.01). These defects in cell adhesion and focal adhesion formation were rescued by re-expression of wild type THSD1 ( p <0.05). In contrast, initial studies indicate that expression of mutated versions of THSD1 as seen in human patients (L5F, R450*, E466G, P639L) could not restore cell adhesion and focal adhesion formation to wild type levels. Conclusions: Our studies provide evidence for a role of THSD1 and THSD1 mutations in endothelial cell adhesion and suggest a possible mechanism underlying THSD1 -mediated aneurysm disease.

scholarly journals Correction to: Matrix metalloproteinase 9 induces endothelial-mesenchymal transition via Notch activation in human kidney glomerular endothelial cells

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Ye Zhao ◽  
Xi Qiao ◽  
Lihua Wang ◽  
Tian Kui Tan ◽  
Hong Zhao ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Matrix Metalloproteinase ◽  
Human Kidney ◽  
Matrix Metalloproteinase 9 ◽  
Mesenchymal Transition ◽  
Glomerular Endothelial Cells ◽  
Metalloproteinase 9 ◽  
Endothelial Mesenchymal Transition ◽  
Notch Activation

An amendment to this paper has been published and can be accessed via the original article.

ANGPTL4 modulates vascular junction integrity by integrin signaling and disruption of intercellular VE-cadherin and claudin-5 clusters

Blood ◽  
2011 ◽  
Vol 118 (14) ◽  
pp. 3990-4002 ◽  
Author(s):  
Royston-Luke Huang ◽  
Ziqiang Teo ◽  
Han Chung Chong ◽  
Pengcheng Zhu ◽  
Ming Jie Tan ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Integrin Signaling ◽  
Wild Type ◽  
Integrin Α5β1 ◽  
Vascular Integrity ◽  
Vascular Disruption ◽  
Binding Partners ◽  
Adhesive Proteins

Abstract Vascular disruption induced by interactions between tumor-secreted permeability factors and adhesive proteins on endothelial cells facilitates metastasis. The role of tumor-secreted C-terminal fibrinogen-like domain of angiopoietin-like 4 (cANGPTL4) in vascular leakiness and metastasis is controversial because of the lack of understanding of how cANGPTL4 modulates vascular integrity. Here, we show that cANGPTL4 instigated the disruption of endothelial continuity by directly interacting with 3 novel binding partners, integrin α5β1, VE-cadherin, and claudin-5, in a temporally sequential manner, thus facilitating metastasis. We showed that cANGPTL4 binds and activates integrin α5β1-mediated Rac1/PAK signaling to weaken cell–cell contacts. cANGPTL4 subsequently associated with and declustered VE-cadherin and claudin-5, leading to endothelial disruption. Interfering with the formation of these cANGPTL4 complexes delayed vascular disruption. In vivo vascular permeability and metastatic assays performed using ANGPTL4-knockout and wild-type mice injected with either control or ANGPTL4-knockdown tumors confirmed that cANGPTL4 induced vascular leakiness and facilitated lung metastasis in mice. Thus, our findings elucidate how cANGPTL4 induces endothelial disruption. Our findings have direct implications for targeting cANGPTL4 to treat cancer and other vascular pathologies.

P-Selectin and Platelet Clearance

Blood ◽  
1998 ◽  
Vol 92 (11) ◽  
pp. 4446-4452 ◽  
Author(s):  
Gaëtan Berger ◽  
Daqing W. Hartwell ◽  
Denisa D. Wagner
Keyword(s):  
Flow Cytometry ◽  
Endothelial Cells ◽  
Platelet Activation ◽  
Soluble Form ◽  
Wild Type ◽  
Adhesion Receptor ◽  
Activated Platelets ◽  
Deficient Mice

P-selectin is an adhesion receptor for leukocytes expressed by activated platelets and endothelial cells. To assess a possible role of P-selectin in platelet clearance, we adapted an in vivo biotinylation technique in mice. Wild-type and P-selectin–deficient mice were infused with N-hydroxysuccinimido biotin. The survival of biotinylated platelets was followed by flow cytometry after labeling with fluorescent streptavidin. Both wild-type and P-selectin–deficient platelets presented identical life spans of about 4.7 days, suggesting that P-selectin does not play a role in platelet turnover. When biotinylated platelets were isolated, activated with thrombin, and reinjected into mice, the rate of platelet clearance was unchanged. In contrast, storage of platelets at 4°C caused a significant reduction in their life span in vivo but again no significant differences were observed between the two genotypes. The infused thrombin-activated platelets rapidly lost their surface P-selectin in circulation, and this loss was accompanied by the simultaneous appearance of a 100-kD P-selectin fragment in the plasma. This observation suggests that the platelet membrane P-selectin was shed by cleavage. In conclusion, this study shows that P-selectin, despite its binding to leukocytes, does not mediate platelet clearance. However, the generation of a soluble form of P-selectin on platelet activation may have biological implications in modulating leukocyte recruitment or thrombus growth.

Endothelial cell P-selectin mediates a proinflammatory and prothrombogenic phenotype in cerebral venules of sickle cell transgenic mice

2004 ◽  
Vol 286 (5) ◽  
pp. H1608-H1614 ◽  
Author(s):  
Katherine C. Wood ◽  
Robert P. Hebbel ◽  
D. Neil Granger
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Sickle Cell ◽  
Leukocyte Adhesion ◽  
Chimeric Mice ◽  
Wild Type ◽  
Cell Disease ◽  
Adhesion Of Platelets

Whereas the adhesion of leukocytes and erythrocytes to vascular endothelium has been implicated in the vasooclusive events associated with sickle cell disease, the role of platelet-vessel wall interactions in this process remains undefined. The objectives of this study were to: 1) determine whether the adhesion of platelets and leukocytes in cerebral venules differs between sickle cell transgenic (βS) mice and their wild-type (WT) counterparts (C57Bl/6) under both resting and posthypoxic conditions, and 2) define the contributions of P-selectin to these adhesion processes. Animals were anesthetized, and platelet and leukocyte interactions with endothelial cells of cerebral postcapillary venules were monitored and quantified using intravital fluorescence microscopy in WT, βS, and chimeric mice produced by transplanting bone marrow from WT or βSmice into WT or P-selectin-deficient (P-sel–/–) mice. Platelet and leukocyte adhesion to endothelial cells in both unstimulated and posthypoxic βSmice were significantly elevated over WT levels. Chimeric mice involving bone marrow transfer from βSmice to P-sel–/–mice exhibited a profound attenuation of both platelet and leukocyte adhesion compared with βSbone marrow transfer to WT mice. These findings indicate that βSmice assume both an inflammatory and prothrombogenic phenotype, with endothelial cell P-selectin playing a major role in mediating these microvascular responses.

scholarly journals Chronic obstructive uropathy in severe combined immunodeficient (SCID) mice: lymphocyte infiltration is not required for progressive tubulointerstitial injury.

1998 ◽  
Vol 9 (6) ◽  
pp. 1008-1017
Author(s):  
S B Shappell ◽  
T Gurpinar ◽  
J Lechago ◽  
W N Suki ◽  
L D Truong
Keyword(s):  
Interstitial Fibrosis ◽  
Obstructive Uropathy ◽  
Inflammatory Cells ◽  
Scid Mice ◽  
Chronic Obstructive ◽  
Lymphocyte Infiltration ◽  
Wild Type ◽  
Tubulointerstitial Injury ◽  
Tubular Atrophy

Progressive renal injury in humans and experimental animal models is characterized by tubular atrophy, infiltration of mononuclear inflammatory cells, and interstitial fibrosis. Permanent unilateral ureter ligation represents a reproducible model for investigating mechanisms of progressive kidney injury, and in the rat is characterized by tubular epithelial cell proliferation followed by apoptosis and progressive infiltration of monocytes and lymphocytes. Nevertheless, whether monocytes or lymphocytes play a dominant role in causing tubulointerstitial damage remains to be elucidated. In the current study, a model of chronic obstructive uropathy in the mouse is established and the role of lymphocyte infiltration in the evolution of the tubule and interstitial alterations is investigated. Permanent ligation of the left ureter in wild-type (C3H/HeJ) mice resulted in progressive atrophy of tubules and interstitial fibrosis compared with the contralateral kidney over a 30-d period. Immunoperoxidase studies on frozen sections taken from kidneys at 0, 3, 10, 20, and 30 d after ureter ligation showed that the tubulointerstitial injury was accompanied by a marked and progressive increase in interstitial macrophages and T lymphocytes, with no appreciable increase in B lymphocytes. No increase in inflammatory cells was detected in contralateral kidneys over the same time frame. The significance of T lymphocyte infiltration was examined by comparing the degree of tubular atrophy and interstitial fibrosis and the nature and quantity of the inflammatory infiltrate in wild-type mice and C3HSMn.C-Scid/J (SCID) mice subjected to permanent left ureter ligation. SCID mice have genetic defects in immunoglobulin and T cell receptor gene rearrangements and are devoid of circulating mature B and T lymphocytes. Wild-type and SCID mice developed tubular atrophy and interstitial volume expansion in the ligated kidney to the same degree and at the same rate. SCID mice developed a prominent and marked monocyte/macrophage infiltrate in the ligated kidney, which was essentially equal to that in wild-type mice. In contrast, consistent with the known absence of mature lymphocytes in SCID mice, there was essentially no T lymphocyte infiltration into the ligated kidney of SCID mice. These results demonstrate the effective establishment of the model of maintained unilateral ureter ligation in mice, which is readily applicable to genetic mutant strains thus allowing for specific investigation of the role of individual components of the inflammatory response in progressive tubulointerstitial injury. These studies further demonstrate that lymphocyte infiltration is not required for progressive tubular atrophy and increased interstitial fibrosis after maintained unilateral ureter ligation.

Abstract 097: Knockout of Matrix Metalloproteinase 9 Protects Against Hypertension-induced Renal Disease in Hypertensive Dahl S Rats

Hypertension ◽  
2017 ◽  
Vol 70 (suppl_1) ◽  
Author(s):  
Chao Zhang ◽  
Paige N Mims ◽  
Tongyu Zhu ◽  
Fan Fan ◽  
Richard J Roman
Keyword(s):  
Matrix Metalloproteinase ◽  
Diastolic Pressure ◽  
Interstitial Fibrosis ◽  
Systolic Pressure ◽  
Vascular Wall ◽  
Matrix Metalloproteinase 9 ◽  
Glomerular Injury ◽  
Hypertensive Nephropathy ◽  
Metalloproteinase 9

Matrix metalloproteinase 9 (MMP9) is a member of gelatinase family of enzymes with potentially opposing actions on tissue fibrosis (i.e. the degradation of extracellular matrix). Our previous study showed that two nonselective MMP inhibitors, XL081 and XL784, which inhibit MMP2, 9, 13 and Adam10 could delay and even reverse the development of renal fibrosis in hypertensive Dahl Salt sensitive (SS) rats. However, the specific role of MMP9 to the development of hypertensive nephropathy in MMP9 knockout (KO) rat models is unknown. In the present study, MMP9 KO rats were created on the SS genetic background using a CRISPR/Cas 9 system and the effect of KO was verified in this model. The systolic pressure, diastolic pressure and mean blood pressure (MAP) were similar in 12 weeks old MMP9 KO (n=6) and SS rats (n=9) fed a low salt diet (145 ± 3 vs. 148 ± 2mmHg; 111 ± 2 vs. 107 ± 2mmHg; 128 ± 2 vs. 126 ± 2mmHg). MAP increased to 180 ± 4 vs. 154 ± 3mmHg in SS rats versus MMP9 KO rats fed 8% high salt (HS) diet for 3 weeks. Proteinuria increased from 196 ± 18 mg/day to 718 ± 61mg/day in HS treated SS rats (n=9). It was significantly reduced in HS treated MMP9 KO rats (211 ± 30mg/day, n=6). The degree of glomerular injury (2.88 ± 0.08 vs. 3.52 ± 0.02), interstitial fibrosis (4.57 ± 0.35% vs. 10.45 ± 0.55%), vascular wall-to-lumen ratio (0.63 ± 0.04 vs. 1.22 ± 0.08) and protein cast area (6.40 ± 0.07% vs. 20.27 ± 2.65%) were all significantly reduced in MMP9 KO rats (n=6) versus the corresponding values in SS rats (n=6) fed 8% HS diet for 3 weeks. Autoregulation of renal blood flow (RBF) to elevations in perfusion was impaired in SS rats prior to the development of hypertension, for RBF rose by 20.6 ± 3.6% (n=8) when MAP was increased from 110 to 150 mmHg. Autoregulation of RBF was restored in MMP9 KO rats and only increased by 7% when pressure was increased over the same range. In contrast, there was no difference in the fall in RBF in SS versus MMP9 KO rats when pressure was reduced from 110 to 50 mmHg. These findings suggest that knockout of MMP9 in SS rats restores autoregulation of RBF and opposes the development of hypertension, proteinuria, glomerular injury and renal interstitial fibrosis.

Abstract 3662: Involvement Of The Small Gtpase Rap1 In Integrin Signaling In Endothelial Cells And Postnatal Neovascularization

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Guillaume Carmona ◽  
Alessia Orlandi ◽  
Henschler Reinhard ◽  
Andreas. M Zeiher ◽  
Stefanie Dimmeler ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Microvessel Density ◽  
Umbilical Vein ◽  
Critical Role ◽  
Significant Inhibition ◽  
Small Gtpase ◽  
Integrin Signaling ◽  
Wild Type

Ras associated protein 1 (Rap1), a small GTPase of the Ras family, has attracted much attention because of its involvement in several aspects of cell adhesion, including integrin- and cadherin-mediated adhesion. Yet, the role of Rap1 genes for integrin signaling in endothelial cells (EC) and angiogenesis has not been investigated. Human umbilical vein endothelial cells (HUVEC) express Rap1a and Rap1b mRNA as assessed by RT-PCR. In order to determine the contribution of Rap1 activity for angiogenesis, we overexpressed Rap1GAP1, a GTPase activating protein, which specifically inhibits the activity of both Rap1a and Rap1b. Overexpression of Rap1GAP1 led to a significant inhibition of angiogenic sprouting of HUVEC under basal conditions and bFGF stimulation by 44 ± 5 % in a 3-dimensional spheroidal system and blocked tube formation in a matrigel assay, migration and adhesion. In order to separately investigate the role of Rap1a and Rap1b genes in angiogenesis, we performed gene silencing with siRNA. Silencing of either Rap1a or Rap1b significantly and additively blocked the sprouting of HUVEC under basal and bFGF-stimulated conditions (Rap1a-siRNA: 55 ± 5 %, Rap1b-siRNA: 61 ± 9 % and Rap1a+Rap1b siRNA: 73 ± 5% inhibition) and significantly reduced HUVEC migration and adhesion on fibronectin and collagen. Moreover, silencing of Rap1a and Rap1b reduced beta1-integrin affinity in HUVEC, suggesting the importance of Rap1a and Rap1b for inside-out integrin activation in EC. In addition, silencing of Rap1a and Rap1b prevented VEGF-induced PKB/Akt1 activation. These data prompted us to investigate the in vivo role of Rap1a using Rap1a-deficient mice. Interestingly, Rap1a −/− mice are born with a substantially reduced mendelian ratio. Rap1a +/− heterozygote mice displayed decreased microvessel density in comparison to wild-type mice (Rap1a +/+ ) in a matrigel plug assay. Moreover Rap1a +/− and Rap1a −/− displayed significantly reduced microvessel density in ischemic muscles in the model of hind limb ischemia in comparison to wild-type mice (Rap1a +/− : 32 ± 3 % ; Rap1a −/− : 43 ± 3 % inhibition). Thus, our data demonstrated a critical role of Rap1 in the regulation of β1-integrin signaling in endothelial cells and for postnatal neovascularization.

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