Endothelial‐to‐mesenchymal transition (EndMT) was first reported in the embryogenesis. Recent studies show that EndMT also occurs in the disease progression of atherosclerosis, cardiac and pulmonary ...fibrosis, pulmonary hypertension, diabetic nephropathy, and cancer. Although transforming growth factor β (TGFβ) is crucial for EndMT, it is not clear which isoform elicits a predominant effect. The current study aims to directly compare the dose‐dependent effects of TGFβ1, TGFβ2, and TGFβ3 on EndMT and characterize the underlying mechanisms. In our results, all three TGFβ isoforms induced EndMT in human microvascular endothelial cells after 72 hr, as evidenced by the increased expression of mesenchymal markers N‐cadherin and α‐smooth muscle actin as well as the decreased expression of endothelial nitric oxide synthase. Interestingly, the effect of TGFβ2 was the most pronounced. At 1 ng/ml, only TGFβ2 treatment resulted in significantly increased phosphorylation (activation) of Smad2/3 and p38‐MAPK and increased expression of mesenchymal transcription factors Snail and FoxC2. Intriguingly, we observed that treatment with 1 ng/ml TGFβ1 and TGFβ3, but not TGFβ2, resulted in an increased expression of TGFβ2, thus indicating that EndMT with TGFβ1 and TGFβ3 treatments was due to the secondary effects through TGFβ2 secretion. Furthermore, silencing TGFβ2 using small interfering RNA blunted the expression of EndMT markers in TGFβ1‐ and TGFβ3‐treated cells. Together, our results indicate that TGFβ2 is the most potent inducer of EndMT and that TGFβ1‐ and TGFβ3‐induced EndMT necessitates a paracrine loop involving TGFβ2.
Our study demonstrates that transforming growth factor β2 (TGFβ2) is the most potent inducer of endothelial‐to‐mesenchymal transition (EndMT) and that TGFβ1‐ and TGFβ3‐induced EndMT necessitates a paracrine loop involving TGFβ2. Silencing TGFβ2 using small interfering RNA blunted the expression of EndMT markers in TGFβ1‐ and TGFβ3‐treated cells.
Display omitted
Decades of research have elucidated the critical role of Akt isoforms in cancer as pro-tumorigenic and metastatic regulators through their specific effects on the cancer cells, tumor ...endothelial cells and the stromal cells. The pro-cancerous role of Akt isoforms through enhanced cell proliferation and suppression of apoptosis in cancer cells and the cells in the tumor microenvironment is considered a dogma. Intriguingly, studies also indicate that the Akt pathway is essential to protect the endothelial-barrier and prevent aberrant vascular permeability, which is also integral to tumor perfusion and metastasis. To complicate this further, a flurry of recent reports strongly indicates the metastasis suppressive role of Akt, Akt1 in particular in various cancer types. These reports emanated from different laboratories have elegantly demonstrated the paradoxical effect of Akt1 on cancer cell epithelial-to-mesenchymal transition, invasion, tumor endothelial-barrier disruption, and cancer metastasis. Here, we emphasize on the specific role of Akt1 in mediating tumor cell-vasculature reciprocity during the advanced stages of cancers and discuss how Akt1 differentially regulates cancer metastasis through mechanisms distinct from its pro-tumorigenic effects. Since Akt is integral for insulin signaling, endothelial function, and metabolic regulation, we also attempt to shed some light on the specific effects of diabetes in modulating Akt pathway in the promotion of tumor growth and metastasis.
The antioxidant potency of different fractions of Moringa oleifera leaves were investigated by employing various established in vitro systems, such as β-Carotene bleaching, reducing power, ...DPPH/superoxide/hydroxyl radical scavenging, ferrous ion chelation and lipid peroxidation. On the basis of in vitro antioxidant properties polyphenolic fraction of M. oleifera leaves (MOEF) was chosen as the potent fraction and used for the DNA nicking and in vivo antioxidant properties. MOEF shows concentration dependent protection of oxidative DNA damage induced by HO and also found to inhibit the toxicity produced by CCl4 administration as seen from the decreased lipid peroxides (LPO) and increased glutathione (GSH) levels. Among the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) levels were restored to almost normal levels compared to CCl4 intoxicated rats. The HPLC analysis indicated the presence of phenolic acids (gallic, chlorogenic, ellagic and ferulic acid) and flavonoids (kaempferol, quercetin and rutin). Thus, it may be concluded that the MOEF possess high phenolic content and potent antioxidant properties, which may be mediated through direct trapping of the free radicals and also through metal chelation.
The suppression of Akt1 activity by LPS in pulmonary endothelial cells results in increased FoxO1/3a activation, in turn, leading to increased stromelysin1 expression/activity and reduced expression ...of tight junction proteins, particularly claudin5. Pharmacologically targeting FoxO or stromelysin activity inhibits LPS-induced pulmonary vascular injury and inflammation.
Display omitted
Enhanced vascular permeability is associated with inflammation and edema in alveoli during the exudative phase of acute respiratory distress syndrome (ARDS). Mechanisms leading to the endothelial contribution on the early exudative stage of ARDS are not precise. We hypothesized that modulation of endothelial stromelysin1 expression and activity by Akt1-forkhead box-O transcription factors 1/3a (FoxO1/3a) pathway could play a significant role in regulating pulmonary edema during the initial stages of acute lung injury (ALI). We utilized lipopolysaccharide (LPS)-induced mouse ALI model in vivo and endothelial barrier resistance measurements in vitro to determine the specific role of the endothelial Akt1-FoxO1/3a-stromelysin1 pathway in ALI. LPS treatment of human pulmonary endothelial cells resulted in increased stromelysin1 and reduced tight junction claudin5 involving FoxO1/3a, associated with decreased trans-endothelial barrier resistance as determined by electric cell-substrate impedance sensing technology. In vivo, LPS-induced lung edema was significantly higher in endothelial Akt1 knockdown (EC-Akt1–/–) compared to wild-type mice, which was reversed upon treatment with FoxO inhibitor (AS1842856), stromelysin1 inhibitor (UK356618) or with shRNA-mediated FoxO1/3a depletion in the mouse lungs. Overall, our study provides the hope that targeting FoxO and styromelysin1 could be beneficial in the treatment of ALI.
Cancer-associated fibroblasts (CAFs) are the most abundant stromal cell type in the tumor microenvironment. CAFs orchestrate tumor-stromal interactions, and contribute to cancer cell growth, ...metastasis, extracellular matrix (ECM) remodeling, angiogenesis, immunomodulation, and chemoresistance. However, CAFs have not been successfully targeted for the treatment of cancer. The current study elucidates the significance of glypican-1 (GPC-1), a heparan sulfate proteoglycan, in regulating the activation of human bone marrow-derived stromal cells (BSCs) of fibroblast lineage (HS-5). GPC-1 inhibition changed HS-5 cellular and nuclear morphology, and increased cell migration and contractility. GPC-1 inhibition also increased pro-inflammatory signaling and CAF marker expression. GPC-1 induced an activated fibroblast phenotype when HS-5 cells were exposed to prostate cancer cell conditioned media (CCM). Further, treatment of human bone-derived prostate cancer cells (PC-3) with CCM from HS-5 cells exhibiting GPC-1 loss increased prostate cancer cell aggressiveness. Finally, GPC-1 was expressed in mouse tibia bone cells and present during bone loss induced by mouse prostate cancer cells in a murine prostate cancer bone model. These data demonstrate that GPC-1 partially regulates the intrinsic and extrinsic phenotype of human BSCs and transformation into activated fibroblasts, identify novel functions of GPC-1, and suggest that GPC-1 expression in BSCs exerts inhibitory paracrine effects on the prostate cancer cells. This supports the hypothesis that GPC-1 may be a novel pharmacological target for developing anti-CAF therapeutics to control cancer.
The leaf essential oils from seven Himalayan Lauraceae species viz.
Neolitsea pallens, Lindera pulcherrima, Dodecadenia grandiflora, Persea duthiei, Persea odoratissima, Persea gamblei and
Phoebe ...lanceolata exhibited potent antioxidant and antibacterial activities. The
in vitro antioxidant activity was assessed by using β-carotene bleaching assay, reducing power, DPPH radical scavenging and inhibition of lipid peroxidation methods. The oils of
D. grandiflora and
L.
pulcherrima showed a potent free radical scavenging activity as evidenced by low IC
50 value for DPPH radical (0.032
mg/ml and 0.087
mg/ml, respectively) and inhibition of lipid peroxidation (in between IC
50
=
0.44
mg/ml and IC
50
=
0.74
mg/ml, respectively). The oils were tested against three Gram negative (
Escherichia coli,
Salmonella enterica enterica and
Pasturella multocida) and one Gram positive (
Staphylococcus aureus) bacteria at different concentrations using disc diffusion and tube dilution methods. The inhibition zones (IZ) and MIC values for bacterial strains were in the range of 8.7–22.0
mm and 3.90–31.25
μl/ml, respectively.
Akt1 suppression in advanced cancers has been indicated to promote metastasis. Our understanding of how Akt1 orchestrates this is incomplete. Using the NanoString
-based miRNA and mRNA profiling of ...PC3 and DU145 cells, and subsequent data analysis using the DIANA-mirPath, dbEMT, nCounter, and Ingenuity
databases, we identified the miRNAs and associated genes responsible for Akt1-mediated prostate cancer (PCa) epithelial-to-mesenchymal transition (EMT). Akt1 loss in PC3 and DU145 cells primarily induced changes in the miRNAs and mRNAs regulating EMT genes. These include increased miR-199a-5p and decreased let-7a-5p expression associated with increased TGFβ-R1 expression. Treatment with locked nucleic acid (LNA) miR-199a-5p inhibitor and/or let-7a-5p mimic induced expression changes in EMT genes correlating to their anticipated effects on PC3 and DU145 cell motility, invasion, and TGFβ-R1 expression. A correlation between increased miR-199a-5p and TGFβ-R1 expression with reduced let-7a-5p was also observed in high Gleason score PCa patients in the cBioportal database analysis. Collectively, our studies show the effect of Akt1 suppression in advanced PCa on EMT modulating miRNA and mRNA expression changes and highlight the potential benefits of miR-199a-5p and let-7a-5p in therapy and/or early screening of mPCa.
This study evaluated the antioxidant properties of
Ficus glomerata fruits using
in vitro and
in vivo assay. In order to find
in vitro antioxidant properties, extract/fractions from
F. glomerata were ...studied for TPC, AOA, RP, DPPH
,
O
2
-
,
OH scavenging activities and LPO. Among all the extract/fractions, FEF has shown potent antioxidant activity and was also found effective in protecting oxidative DNA damage. The
in vivo evaluation of oxidative stress (LPO) and antioxidant defenses (concentration of GSH, as well as CAT and SOD activities) were measured in CCl
4 induced toxic rats. FEF was found to inhibit the toxicity as seen from the decreased LPO and increased GSH, SOD and CAT levels. FEF has higher phenolic content and showed the presence of gallic, chlorogenic and ellagic acid. Based on these results, it is concluded that
F. glomerata protects tissues from oxidative stress and these effects are probably related to the antioxidant properties.
The multi-gene claudin (
) family of tight junction proteins have isoform-specific roles in blood-tissue barrier regulation.
, a putative anion pore-forming
based on its structural characterization, ...is assumed to regulate anion balance across the blood-tissue barriers. However, our knowledge about
in physiology and pathology is limited. The current study investigated how
deficiency in mice affects blood electrolytes and kidney structure.
mice revealed no breeding abnormalities, but the newborn pups exhibited delayed growth. Adult
mice displayed electrolyte imbalance, oxidative stress, and injury to the kidneys. Ingenuity pathway analysis followed by RNA-sequencing revealed hyperactivation of signaling pathways and downregulation of SOD1 expression in kidneys associated with inflammation and reactive oxygen species generation, demonstrating the importance of
in the maintenance of electrolytes and reactive oxygen species across the blood-tissue barrier.
Traditionally fruits of
Ficus glomerata Roxb (Family: Moraceae) are used to treat anemia and gastrointestinal disorders.
Gastroprotective effect of 50% ethanolic extract of
F. glomerata fruit (FGE) ...was studied in different gastric ulcer models in rats.
FGE (50, 100 and 200
mg/kg body weight) was administered orally, twice daily for 5 days for prevention from pylorus ligation (PL), ethanol (EtOH) and cold restraint stress (CRS) – induced ulcers. Estimation of H
+K
+ATPase activity and gastric wall mucous were performed in EtOH-induced ulcer and antioxidant enzyme activities in supernatant mitochondrial fraction of CRS-induced ulcers.
FGE showed dose dependent inhibition of ulcer index in pylorus ligation, ethanol and cold restraint stress – induced ulcers. FGE prevents the oxidative damage of gastric mucosa by blocking lipid peroxidation and by significant decrease in superoxide dismutase, H
+K
+ATPase and increase in catalase activity. High performance thin layer chromatography (HPTLC) analysis showed the presence of 0.57% and 0.36% w/w of gallic acid and ellagic acid in FGE.
Our results show that
F. glomerata possess significant gastroprotective activity which might be due to gastric defence factors and phenolics might be the main constituents responsible for this activity.
PDF
