Endothelial Cells ; metabolism ; physiology ; Stem Cells ; metabolism ; physiology

Animals ; Endothelial Cells ; metabolism ; physiology ; Fibroblasts ; metabolism ; physiology ; Humans ; Hypertension, Pulmonary ; etiology ; genetics ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; physiology

Hypoxia-inducible factor (HIF-1α), a core transcription factor responding to changes in cellular oxygen levels, is closely associated with a wide range of physiological and pathological conditions. However, its differential impacts on vascular cell types and molecular programs modulating human vascular homeostasis and regeneration remain largely elusive. Here, we applied CRISPR/Cas9-mediated gene editing of human embryonic stem cells and directed differentiation to generate HIF-1α-deficient human vascular cells including vascular endothelial cells, vascular smooth muscle cells, and mesenchymal stem cells (MSCs), as a platform for discovering cell type-specific hypoxia-induced response mechanisms. Through comparative molecular profiling across cell types under normoxic and hypoxic conditions, we provide insight into the indispensable role of HIF-1α in the promotion of ischemic vascular regeneration. We found human MSCs to be the vascular cell type most susceptible to HIF-1α deficiency, and that transcriptional inactivation of ANKZF1, an effector of HIF-1α, impaired pro-angiogenic processes. Altogether, our findings deepen the understanding of HIF-1α in human angiogenesis and support further explorations of novel therapeutic strategies of vascular regeneration against ischemic damage.
Humans ; Vascular Endothelial Growth Factor A/metabolism* ; Endothelial Cells/metabolism* ; Transcription Factors/metabolism* ; Gene Expression Regulation ; Hypoxia/metabolism* ; Cell Hypoxia/physiology*

OBJECTIVETo evaluate the effect of tissue factor (TF) in extravascular migration of fibrosarcoma cells and hematogenous metastasis.

METHODSThe expression of tissue factor in fibrosarcoma HT1080 cells was analyzed by flow cytometry. The extravascular migration of fibrosarcoma cells was observed in a constructed monolayer vascular endothelial cells and extra-cellular matrix model.

RESULTTissue factor was highly expressed in HT1080 cells. HT1080 migrated and passed through the monolayer vascular endothelial cells to the collagen gel in a time-dependent manner. Anti-TF antibody inhibited extravascular migration of fibrosarcoma cells and the inhibition was concentration-dependent (P<0.05).

CONCLUSIONTissue factor may enhance hematogenous metastasis through extravascular migration of fibrosarcoma cells.

Cell Movement ; Cells, Cultured ; Endothelial Cells ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Neoplasm Metastasis ; Thromboplastin ; metabolism ; physiology

Vascular wall-resident stem cells (VW-SCs) play a critical role in maintaining normal vascular function and regulating vascular repair. Understanding the basic functional characteristics of the VW-SCs will facilitate the study of their regulation and potential therapeutic applications. The aim of this study was to establish a stable method for the isolation, culture, and validation of the CD34⁺ VW-SCs from mice, and to provide abundant and reliable cell sources for further study of the mechanisms involved in proliferation, migration and differentiation of the VW-SCs under various physiological and pathological conditions. The vascular wall cells of mouse aortic adventitia and mesenteric artery were obtained by the method of tissue block attachment and purified by magnetic microbead sorting and flow cytometry to obtain the CD34⁺ VW-SCs. Cell immunofluorescence staining was performed to detect the stem cell markers (CD34, Flk-1, c-kit, Sca-1), smooth muscle markers (SM22, SM MHC), endothelial marker (CD31), and intranuclear division proliferation-related protein (Ki-67). To verify the multipotency of the isolated CD34⁺ VW-SCs, endothelial differentiation medium EBM-2 and fibroblast differentiation medium FM-2 were used. After culture for 7 days and 3 days respectively, endothelial cell markers and fibroblast markers of the differentiated cells were evaluated by immunofluorescence staining and q-PCR. Furthermore, the intracellular Ca²⁺ release and extracellular Ca²⁺ entry signaling were evaluated by TILLvisION system in Fura-2/AM loaded cells. The results showed that: (1) High purity (more than 90%) CD34⁺ VW-SCs from aortic adventitia and mesenteric artery of mice were harvested by means of tissue block attachment method and magnetic microbead sorting; (2) CD34⁺ VW-SCs were able to differentiate into endothelial cells and fibroblasts in vitro; (3) Caffeine and ATP significantly activated intracellular Ca²⁺ release from endoplasmic reticulum of CD34⁺ VW-SCs. Store-operated Ca²⁺ entry (SOCE) was activated by using thapsigargin (TG) applied in Ca²⁺-free/Ca²⁺ reintroduction protocol. This study successfully established a stable and efficient method for isolation, culture and validation of the CD34⁺ VW-SCs from mice, which provides an ideal VW-SCs sources for the further study of cardiovascular diseases.
Mice ; Animals ; Endothelial Cells ; Cell Differentiation/physiology* ; Stem Cells ; Adventitia ; Fibroblasts ; Cells, Cultured ; Antigens, CD34/metabolism*

OBJECTIVETo evaluate the effect of aging on the endothelial function of the penile corpus cavernosum in rats.

METHODSThe intracavernosal pressure (ICP) was compared in response to acetylcholine (Ach, endothelium-dependent vasodilator), sodium nitroprusside (SNP, an NO donor) and A23187 (a calcium ionophore)in the young (5 months old) and aged (20 months old) rats. In addition, the activity of nitric oxide synthase (NOS)in penile cavernosal tissues was examined.

RESULTSAch-mediated ICP was significantly attenuated from the maximum of (54.8 +/- 4.2) in the young rats to (40.3 +/- 2.8) mm Hg in the aged ones (Ach = 0.1 mmol/L), P < 0.01. The ICP to SNP (0.1 mmol/L) was (58.9 +/- 4.7) mm Hg in the young rats and (51.7 +/- 5.3) mm Hg in the aged. No statistically significant difference was noted between the two groups, P > 0.05. The Ach-mediated ICP in the young rats was not significantly augmented from the maximum of (54.8 +/- 4.2) to (55.8 +/- 4.7) mm Hg in the presence of the calcium ionophore A23187 (10 micromol/L), P > 0.05. However, A23187 significantly augmented Ach-mediated ICP in the aged from the maximum of (40.3 +/- 2.8) to (56.2 +/- 4.1) mm Hg, P < 0.01. Finally the activity of nitric oxide synthase was not significantly attenuated in either the aged or the young, P > 0.05.

CONCLUSIONThe endothelial function of the penile cavernosum declines with the advance of age. And endothelial dysfunction may play some role in the mechanisms of age-related erectile dysfunction.

Aging ; physiology ; Animals ; Endothelial Cells ; physiology ; Male ; Nitric Oxide Synthase ; metabolism ; Penis ; cytology ; metabolism ; physiology ; Rats ; Rats, Wistar

Portal hypertension (PHT) is associated with changes in the intrahepatic, systemic and portosystemic collateral circulations. Alteration in vasoreactivity (vasodilation and vasoconstriction) plays a central role in the pathogenesis of PHT by contributing to increased intrahepatic resistance, hyperdynamic circulation and the expansion of the collateral circulation. PHT is also importantly characterized by changes in vascular structure; termed vascular remodeling, which is an adaptive response of the vessel wall that occurs in response to chronic changes in the environment such as shear stress. Angiogenesis, the sprouting of new blood vessels, also occurs in PHT, especially in the expansion of the portosystemic collateral circulation. These complementary processes of vasoreactivity, vascular remodeling and angiogenesis represent important targets in the research for the treatment of portal hypertension.
Collateral Circulation/physiology ; Endothelial Cells/metabolism ; Hepatic Stellate Cells/metabolism ; Humans ; Hypertension, Portal/*etiology ; Liver Circulation/physiology ; Vascular Resistance

Hypertrophic scar is a pathological repair result of excessive accumulation of extracellular matrix after skin damage, which affects the appearance and function of patients with varying degrees. The degree of scar formation is directly related to the strength of inflammatory reaction during wound healing, and excessive or prolonged inflammatory response increases the incidence of hypertrophic scars. Interleukin-6 (IL-6) is a pleiotropic cytokine that is involved in regulating the fibrotic network composed of fibroblasts, macrophages, keratinocytes, and vascular endothelial cells, and is closely related to the formation of hypertrophic scars. This article reviews the role of IL-6 and its signaling pathway in hypertrophic scar formation.
Cicatrix, Hypertrophic/pathology* ; Endothelial Cells/metabolism* ; Fibroblasts/metabolism* ; Humans ; Interleukin-6 ; Skin/pathology* ; Wound Healing/physiology*

Caveolin 1 ; genetics ; metabolism ; Cells, Cultured ; Homocysteine ; physiology ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; Humans ; RNA, Messenger ; genetics ; metabolism

Angiotensin II ; pharmacology ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; physiology ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; physiology ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; physiology ; Oxygen ; metabolism