Die Rolle und der Einfluss des Pigment epithelium-derived factor (PEDF) am peripheren Nerventumor, Hirn-Trauma und Schlaganfall

Pigment epithelium-derived factor (PEDF) is a neurotrophic factor with neuroprotective, antiangiogenic, anti-inflammatory, and anti-tumorigenic effects. We aimed to show the effects of PEDF on angiogenesis and tumor growth of malignant peripheral nerve sheath tumor (MPNST) as well as on lesion volume, cell death, cell proliferation, and behavioral outcome after brain injuries (traumatic brain injury and stroke). We used the controlled cortical impact injury (CCI) rat model to study traumatic brain injury and the middle cerebral artery occlusion (MCAO) mouse model for focal cerebral ischemia. In the first study, we demonstrated for the first time that PEDF inhibited proliferation and augmented cell death in S462 MPNST cells after 48 h of treatment in culture. Following transplantation of S462 MPNST cells in athymic nude mice, PEDF reduced MPNST tumor burden mainly due to inhibition of angiogenesis. In the second study, we detected a significant increase of PEDF mRNA levels in post-CCI rat brains. In vivo, PEDF infusion showed no significant decrease in the contusion volume, whereas the number of dead cells, activated microglia, and BrdU-positive cells around the lesion was significantly decreased. In contrast, PEDF infusion significantly increased cell proliferation in the ipsilateral subventricular zone. In the third study, our model produced an ischemic injury confined solely to striatal damage in mice. We detected no reduction in infarct size and cell death in PEDF- vs. cerebrospinal fluid-infused MCAO mice. Behavioral outcome and cell proliferation did not differ between the groups. Whereas PEDF showed a specific positive effect after traumatic brain injury in rats, we were not able to observe the same effect after striatal ischemia in the MCAO mouse model. However, we cannot exclude that PEDF might work under different conditions in stroke. Therefore, further studies will elucidate the effect of PEDF treatment on cell proliferation and outcome in moderate to severe ischemic injury in the brain. Due to its inhibitory effects on the growth of human MPNST, PEDF seems to be promising for future therapeutic purposes against MPNST.Der Pigment epithelium-derived factor (PEDF) ist ein neurotropher Faktor, der Gehirnzellen schützt, die Bildung neuer Gefäßstrukturen und die Entstehung von Tumoren verhindert sowie entzündungshemmend wirkt. Unser Ziel war es, die Auswirkungen von PEDF auf die Gefäßbildung und das Tumorwachstum bei malignen peripheren Nervenscheidentumor (MPNST) sowie auf das Läsionsvolumen, Zelltod, Zellteilung und Verhalten bei Hirnschädigungen (Schädelhirntrauma und Schlaganfall) zu zeigen. Schädelhirntrauma wurde experimentell durch kontrollierte kortikale Kontusion (CCI) in der Ratte herbeigeführt und der Schlaganfall wurde mittels Mausmodell des Verschluss der mittleren Hirnarterie (MCAO) untersucht. In der ersten Studie zeigten wir erstmals, dass sich kultivierte S462 MPNST Zellen, die 48 Stunden lang mit PEDF behandelt wurden, weniger stark teilten und stärker abstarben. Nach Transplantation der S462 MPNST Zellen in athymische Naktmäuse verringerte PEDF außerdem die Belastung durch den MPNST Tumor. Dies war auf eine Verringerung der Bildung neuer Gefäßstrukturen zurückzuführen. In der zweiten Studien stellten wir eine signifikant erhöhte mRNA-Expression von PEDF nach CCI in Rattengehirnen fest. Infusion von PEDF führte zu einer nichtsignifikanten Verringerung des Kontusionsvolumens nach CCI, während sich die Zahl der toten Zellen, aktivierten Microglia und BrdU-positiven Zellen um die Läsion signifikant verringerte. Im Gegensatz dazu war die Zellteilung in der ipsilateralen Suventrikularzone signifikant erhöht. In der dritten Studie führte die MCAO zu einem striatalen Schlaganfall in der Maus. Wir stellten keine Reduktion des Infarktvolumens und des Zelltods nach experimentellem Schlaganfall zwischen Tieren, die eine Infusion von PEDF oder Zerebrospinalflüssigkeit erhielten, fest. Ebenso gab es keine Unterschiede im Verhalten oder der Zellteilung zwischen den beiden Gruppen. Während PEDF einen positiven Effekt nach Schädelhirntrauma in der Ratte zeigte, konnten wir diesen Effekt nach striatalem Schlaganfall in der Maus nicht bestätigen. Jedoch können wir nicht ausschließen, dass PEDF unter anderen Bedingungen auch beim Schlaganfall wirksam sein kann. Weiterführende Studien sind notwendig, um die Auswirkungen der PEDF-Behandlung bei moderatem bis schwerwiegendem Schlaganfall aufzuklären. Aufgrund seines inhibitorischen Effekts auf das Wachstum von humanen MPNSTs, scheint PEDF ein vielversprechender Wirkstoff zur Therapie von MPNST zu sein

In vitro apoptotic and antiproliferative effects of propranolol on human breast cancer cells

326-331Breast cancer is an issue of concern with increasing incidence among women worldwide. Propranolol, as an antihypertensive drug, exerts anticancer effects too. We conducted this study to analyze the in vitro apoptotic and anti-proliferative effects of propranolol in human MCF-7 breast cancer cells. MCF-7 cells were seeded into 6-well plates and treated with 50 μL propranolol for 24 hours. After cell homogenization, the levels of pro-apoptotic proteins BCL2 associated X (BAX), apoptosis inducing factor (AIF), C/-EBP homologous protein (GADD153), and glucose-regulated protein 78 (GRP78), anti-apoptotic protein BCL2 apoptosis regulator (BCL-2), and cycle-regulator WEE1 G2 checkpoint kinase (WEE1) were measured with ELISA. Propranolol significantly upregulated pro-apoptotic proteins AIF, BAX, GADD153, and GRP78 while downregulated anti-apoptotic protein BCL2. The level of WEE1, as the main regulatory cell cycle protein at the G2/M checkpoint, significantly increased after propranolol treatment. Propranolol inhibited the proliferation of MCF-7 human breast cancer cells by upregulating pro-apoptotic factors AIF, BAX, GADD153 and GRP78 and by downregulating antiapoptotic BCL2. Elevated WEE1 levels after propranolol treatment might lead the tumor cells into a sustained cell-cycle arrest which eventually resulted in caspase-dependent or -independent mitochondrial or endoplasmic-reticulum stress-induced apoptosis. So, propranolol can be utilized as a potential therapeutic agent in breast cancer therapy

The Effect of Fingolimod (FTY720) Treatment on Liver Enzyme Levels in Relapsing-Remitting Multiple Sclerosis Patients

Aim: Multiple sclerosis (MS) is a chronic inflammatory pathology affecting the centralnervous system. Many therapeutic options have been approved against MS until today. In thisstudy, it was aimed to investigate the effect of fingolimod treatment (FT) on the liver enzymelevels of relapsing-remitting multiple sclerosis (RRMS) patients.Material and Methods: Body mass index, FT (0.5 mg/day) duration, and liver enzyme(alanine aminotransferase, ALT; gamma glutamyl transferase, GGT) levels of 102 RRMSpatients (66 female, 36 male, mean age was 40.9±10.9 years) were gathered from polyclinicrecords retrospectively.Results: The FT duration of MS patients was between 0.5 and 6 years. Increased ALT andGGT levels were detected in RRMS patients after >3 month-long FT. After FT, ALT and GGTlevels elevated in males almost 2 times higher than in females. It was observed that ALT andGGT levels increased by 1.3 and 1.5 times in females, while 1.6 and 1.9 times in males,respectively. Of the MS patients with increased transaminases post-FT, 7 (23.3%) males and8 (17.4%) females were at upper limit of normal for ALT whereas 9 (34.6%) males and 14(32.6%) females as for GGT. Age and FT duration did not affect ALT and GGT levels.Conclusion: Overall, FT elevated ALT and GGT levels of RRMS patients. Thus, it is of highimportance to monitor MS patients throughout FT. So that, we suggest tracking ALT and GGTlevels during and after FT to prevent possible liver damage or the occurrence of other systemicdiseases

The Effect of Fingolimod (FTY720) Treatment on Liver Enzyme Levels in Relapsing-Remitting Multiple Sclerosis Patients

Aim: Multiple sclerosis (MS) is a chronic inflammatory pathology affecting the centralnervous system. Many therapeutic options have been approved against MS until today. In thisstudy, it was aimed to investigate the effect of fingolimod treatment (FT) on the liver enzymelevels of relapsing-remitting multiple sclerosis (RRMS) patients.Material and Methods: Body mass index, FT (0.5 mg/day) duration, and liver enzyme(alanine aminotransferase, ALT; gamma glutamyl transferase, GGT) levels of 102 RRMSpatients (66 female, 36 male, mean age was 40.9±10.9 years) were gathered from polyclinicrecords retrospectively.Results: The FT duration of MS patients was between 0.5 and 6 years. Increased ALT andGGT levels were detected in RRMS patients after >3 month-long FT. After FT, ALT and GGTlevels elevated in males almost 2 times higher than in females. It was observed that ALT andGGT levels increased by 1.3 and 1.5 times in females, while 1.6 and 1.9 times in males,respectively. Of the MS patients with increased transaminases post-FT, 7 (23.3%) males and8 (17.4%) females were at upper limit of normal for ALT whereas 9 (34.6%) males and 14(32.6%) females as for GGT. Age and FT duration did not affect ALT and GGT levels.Conclusion: Overall, FT elevated ALT and GGT levels of RRMS patients. Thus, it is of highimportance to monitor MS patients throughout FT. So that, we suggest tracking ALT and GGTlevels during and after FT to prevent possible liver damage or the occurrence of other systemicdiseases

Biological behaviors of muscarinic receptors in mesenchymal stem cells derived from human placenta and bone marrow

WOS: 000498871600017Objective(s): Cells perform their functional activities by communicating with each other through endogenous substances and receptors. Post-translation, stem cells function properly in new host tissue by carrying specific cell surface receptors. We aimed to characterize muscarinic receptor subtypes in mesenchymal stem cells (MSCs) together with osteogenic and adipogenic differentiation markers. Materials and Methods: mRNA levels of 5 muscarinic receptor subtypes (CHRM1 to 5), BMP-6, and PPAR gamma during osteogenic and adipogenic differentiation, under the effect of atropine blockade, were measured in MSCs obtained from human fetal membrane (FM) and bone marrow (BM). Additionally, the effect of atropine on differentiation in the 1st, 2nd, and 3rd passages of MSCs, obtained from human FM and BM, were analyzed by RT-qPCR. Results: CHRM1 mRNA levels increased in the FM group, while decreasing in the BM group. We found significant decreases in CHRM3 and CHRM5 mRNA levels in FM and BM groups, respectively. Atropine had variable effects based on cell source and receptor type. BMP-6 mRNA levels in differentiated osteogenic cells increased significantly compared to undifferentiated cells in both FM and BM groups. In MSCs derived from both sources, PPAR gamma mRNA levels in differentiated adipogenic cells increased significantly. Atropine showed no effect on MSCs differentiation. Conclusion: These results indicate that expressions of muscarinic receptors in MSCs derived from BM and FM can vary and these cells keep the potential of osteogenic and adipogenic differentiation in vitro. Besides, atropine had no effect on adipogenic and osteogenic differentiation of MSCs.Cukurova University, Scientific Research Projects Fund, Adana, Turkey [TF2014D2]The results presented in this paper were part of a student thesis. The authors express their great gratitude to Canan Cansun and Nilay Oktar for their invaluable help in the RT-qPCR technique. This study was supported by Cukurova University, Scientific Research Projects Fund, Adana, Turkey (project no: TF2014D2)

Effects of pigment epithelium derived factor (PEDF) on malignant peripheral nerve sheath tumours (MPNSTs)

Neurofibromatosis type 1 (NF1) is an inherited genetic disease affecting 1 in 3,500 individuals. A prominent feature of NF1 is the formation of benign tumours of the peripheral nerve sheath (neurofibromas). However, these can become malignant and form highly metastatic malignant peripheral nerve sheath tumours (MPNST), which are usually fatal despite aggressive surgery, chemotherapy, and radiotherapy. Recent studies have shown that pigment epithelium-derived factor (PEDF) can induce differentiation and inhibit angiogenesis in several kinds of tumours. The present study was designed to determine the in vitro and in vivo effects of PEDF on MPNST angiogenesis and tumour growth. PEDF inhibited proliferation and augmented apoptosis in S462 MPNST cells after 48 h of treatment in culture. In xenografts of S462 MPNST cells in athymic nude mice, PEDF suppressed MPNST tumour burden, due mainly to inhibition of angiogenesis. These results demonstrate for the first time inhibitory effects of PEDF on the growth of human MPNST via induction of anti-angiogenesis and apoptosis. Our results suggest that PEDF could be a novel approach for future therapeutic purposes against MPNST. © 2013 Springer Science+Business Media New York.N

Urotensin-II Prevents Cartilage Degeneration in a Monosodium Iodoacetate-Induced Rat Model of Osteoarthritis

Osteoarthritis (OA) is a common degenerative articular disorder caused by traumatic or spontaneous factors such as genetics, obesity, and advanced age. Comprehending the pathogenic mechanism of OA ensures the development of novel disease-modifying therapeutics rather than conventional palliative drugs with undesired side effects. Urotensin-II (UII) is a multifunctional short cyclic peptide implicated in several disorders. We aimed to analyze the effects of intraarticular UII treatment in a monosodium iodoacetate (MIA)-induced OA rat model. We divided animals into six groups to test three different concentrations of UII with histopathological and immunohistochemical analyses of bone morphogenetic protein-2 (BMP-2), nuclear factor kappa B subunit 1 (NF-kappa B), and intrinsic UII expression. We analyzed serum levels of cartilage related and inflammatory markers post-OA. We observed a noticeable amelioration of the MIA-induced knee damage in UII-treated animals after gross morphology examination. Mankin scoring after histopathological stainings revealed a partial prevention of articular tissue damage in UII-treated animals. We found a significant reduction in BMP-2 and NF-kappa B while an increase in intrinsic UII expressions upon exogenous UII injection after immunohistochemical analyses. The Mankin scores were significantly correlated with BMP-2, NF-kappa B, and intrinsic UII levels. There was no significant alteration in serum markers after UII treatment. We are the first group showing the protective effect of UII on the destructed knee joints of osteoarthritic rats by downregulating the BMP-2 and NF-kappa B and upregulating intrinsic UII expressions. To uncover the mechanistic role of UII during OA, further experiments are warranted. [GRAPHICS]

Pigment Epithelium-Derived Factor Improves Paracellular Blood-Brain Barrier Integrity in the Normal and Ischemic Mouse Brain (vol 16, pg 513, 2020)

Pigment epithelium-derived factor (PEDF) is a neurotrophic factor with neuroprotective, antiangiogenic, and antipermeability effects. In the brain, blood-brain barrier (BBB) function is essential for homeostasis. Its impairment plays a crucial role in the pathophysiology of many neurological diseases, including ischemic stroke. We investigated (a) whether PEDF counteracted vascular endothelial growth factor (VEGF)-induced BBB disruption in the mouse brain, (b) the time course and route of BBB permeability and the dynamics of PEDF expression after cerebral ischemia, and (c) whether intraventricular infusion of PEDF ameliorated brain ischemia by reducing BBB impairment. C57Bl6/N mice received intraparenchymal injections of CSF, VEGF, or a combination of VEGF and PEDF. PEDF increased paracellular but not transcellular BBB integrity as indicated by an increase in the tight junction protein claudin-5. In another group of mice undergoing 60-min middle cerebral artery occlusion (MCAO), transcellular BBB permeability (fibrinogen staining in the absence of a loss of claudin-5) increased as early as 6 h after reperfusion. PEDF immunofluorescence increased at 24 h, which paralleled with a decreased paracellular BBB permeability (claudin-5). PEDF after MCAO originated from the blood stream and endogenous pericytes. In the third experiment, the intraventricular infusion of PEDF decreased edema and cell death after MCAO, potentially mediated by the improvement of the paracellular route of BBB permeability (claudin-5) in the absence of an amelioration of Evans Blue extravasation. Together, our data suggest that PEDF improves BBB function after cerebral ischemia by affecting the paracellular but not the transcellular route. However, further quantitative data of the different routes of BBB permeability will be required to validate our findings

Pigment Epithelium-Derived Factor Improves Paracellular Blood-Brain Barrier Integrity in the Normal and Ischemic Mouse Brain

Pigment epithelium-derived factor (PEDF) is a neurotrophic factor with neuroprotective, antiangiogenic, and antipermeability effects. In the brain, blood-brain barrier (BBB) function is essential for homeostasis. Its impairment plays a crucial role in the pathophysiology of many neurological diseases, including ischemic stroke. We investigated (a) whether PEDF counteracted vascular endothelial growth factor (VEGF)-induced BBB disruption in the mouse brain, (b) the time course and route of BBB permeability and the dynamics of PEDF expression after cerebral ischemia, and (c) whether intraventricular infusion of PEDF ameliorated brain ischemia by reducing BBB impairment. C57Bl6/N mice received intraparenchymal injections of CSF, VEGF, or a combination of VEGF and PEDF. PEDF increased paracellular but not transcellular BBB integrity as indicated by an increase in the tight junction protein claudin-5. In another group of mice undergoing 60-min middle cerebral artery occlusion (MCAO), transcellular BBB permeability (fibrinogen staining in the absence of a loss of claudin-5) increased as early as 6 h after reperfusion. PEDF immunofluorescence increased at 24 h, which paralleled with a decreased paracellular BBB permeability (claudin-5). PEDF after MCAO originated from the blood stream and endogenous pericytes. In the third experiment, the intraventricular infusion of PEDF decreased edema and cell death after MCAO, potentially mediated by the improvement of the paracellular route of BBB permeability (claudin-5) in the absence of an amelioration of Evans Blue extravasation. Together, our data suggest that PEDF improves BBB function after cerebral ischemia by affecting the paracellular but not the transcellular route. However, further quantitative data of the different routes of BBB permeability will be required to validate our findings