scholarly journals High Concentration of C5a-Induced Mitochondria-Dependent Apoptosis in Murine Kidney Endothelial Cells

2019 ◽  
Vol 20 (18) ◽  
pp. 4465 ◽  
Author(s):  
I-Jung Tsai ◽  
Wei-Chou Lin ◽  
Yao-Hsu Yang ◽  
Yu-Lin Tseng ◽  
Yen-Hung Lin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Nephrotic Syndrome ◽  
Cytochrome C ◽  
Idiopathic Nephrotic Syndrome ◽  
Mitochondrial Damage ◽  
High Dose ◽  
High Concentration ◽  
Cytochrome C Release ◽  
Ros Formation ◽  
High Concentrations

Patients with a relapse of idiopathic nephrotic syndrome have significantly increased levels of serum complement component 5a (C5a), and proteinuria has been noted in mice treated with C5a via changes in permeability of kidney endothelial cells (KECs) in established animal models. However, the apoptosis of KECs treated with high concentrations of C5a has also been observed. As mitochondrial damage is known to be important in cell apoptosis, the aim of this study was to examine the association between C5a-induced mouse KEC apoptosis and mitochondrial damage. Mouse KECs were isolated and treated with different concentrations of C5a. Cell viability assays showed that a high-concentration mouse recombinant protein C5a (rmC5a) treatment reduced mouse KEC growth. Cell cycle phase analysis, including apoptosis (sub-G1 phase) showed an increased percentage of the subG1 phase with a high-concentration rmC5a treatment. Cytochrome c and caspase 3/9 activities were significantly induced in the mouse KECs after a high-dose rmC5a (50 ng/mL) treatment, and this was rescued by pretreatment with the C5a receptor (C5aR) inhibitor (W-54011) and N-acetylcysteine (NAC). Reactive oxygen species (ROS) formation was detected in C5a-treated mouse KECs; however, W-54011 or NAC pretreatment inhibited high-dose rmC5a-induced ROS formation and also reduced cytochrome c release, apoptotic cell formation, and apoptotic DNA fragmentation. These factors determined the apoptosis of mouse KECs treated with high-dose C5a through C5aR and subsequently led to apoptosis via ROS regeneration and cytochrome c release. The results showed that high concentrations of C5a induced mouse KEC apoptosis via a C5aR/ROS/mitochondria-dependent pathway. These findings may shed light on the potential mechanism of glomerular sclerosis, a process in idiopathic nephrotic syndrome causing renal function impairment.

scholarly journals Mesalazine Induces Oxidative Stress and Cytochrome c Release in Isolated Rat Heart Mitochondria: An Analysis of Cardiotoxic Effects

2020 ◽  
Vol 39 (3) ◽  
pp. 241-247
Author(s):  
Ahmad Salimi ◽  
Farnaz Bahreini ◽  
Zhaleh Jamali ◽  
Jalal Pourahmad
Keyword(s):  
Cytochrome C ◽  
Rat Heart ◽  
Isolated Rat Heart ◽  
Mitochondrial Swelling ◽  
Heart Mitochondria ◽  
Cytochrome C Release ◽  
Mitochondrial Ros ◽  
Ros Formation ◽  
Rat Heart Mitochondria ◽  
Inflammatory Bowel

Mesalazine is widely used in the management of inflammatory bowel disease. Previous studies reported that mesalazine-induced cardiotoxicity is a rare, potentially fatal complication. Mitochondria play an important role in myocardial tissue homeostasis. Deterioration in mitochondrial function will eventually lead to cardiomyocyte death and consequently cardiovascular dysfunction. The aim of the current study was to investigate the effects of mesalazine on rat heart mitochondria. Rat heart mitochondria were isolated by mechanical lysis and differential centrifugation. Parameters of mitochondrial toxicity including succinate dehydrogenase (SDH) activity, reactive oxygen species (ROS) formation, mitochondrial membrane potential (MMP) collapse, mitochondrial swelling, and cytochrome c release were evaluated. Results revealed that mesalazine induced a concentration- and time-dependent rise in mitochondrial ROS formation, inhibition of SDH, MMP collapse, mitochondrial swelling, and cytochrome c release in rat heart mitochondria. These results indicate that the cardiotoxic effects of mesalazine are most likely associated with mitochondrial dysfunction and ROS formation, which finally ends in cytochrome c release signaling and induction of apoptosis.

scholarly journals Open-Label Clinical Trials of Oral Pulse Dexamethasone for Adults with Idiopathic Nephrotic Syndrome

2019 ◽  
Vol 49 (5) ◽  
pp. 377-385 ◽  
Author(s):  
Monique E. Cho ◽  
Mary H. Branton ◽  
David A. Smith ◽  
Linda Bartlett ◽  
Lilian Howard ◽  
...  
Keyword(s):  
Nephrotic Syndrome ◽  
Adverse Events ◽  
Idiopathic Nephrotic Syndrome ◽  
High Dose ◽  
Open Label ◽  
Serious Adverse Events ◽  
Oral Dexamethasone ◽  
Dose Oral ◽  
Pulse Dexamethasone ◽  
Daily Prednisone

Background: In adults with primary focal segmental glomerulosclerosis (FSGS), daily prednisone may induce complete remissions (CR) and partial remissions (PR), but relapses are frequent and adverse events are common. Methods: We carried out 2 open-label, uncontrolled trials to explore the efficacy and tolerability of pulse oral dexamethasone as an alternative to daily prednisone. We enrolled adult patients with proteinuria > 3.5 g/day despite the use of renin-angiotensin-aldosterone blockade. In the first trial, we enrolled 14 subjects with FSGS and administered 4 dexamethasone doses (25 mg/m2) daily for 4 days, repeated every 28 days over 32 weeks. The second trial involved a more intensive regimen. Eight subjects received 4 dexamethasone doses of 50 mg/m2 every 4 weeks for 12 weeks, followed by 4 doses of 25 mg/m2 every 4 weeks for 36 weeks; subjects were randomized to 2 doses every 2 weeks or 4 doses every 4 weeks. Results: In the first trial, we enrolled 13 subjects with FSGS and 1 with minimal change disease and found a combined CR and PR rate of 36%. In the second trial, we enrolled 8 subjects. The combined CR and PR rate was 29%. Analysis combining both trials showed a combined CR and PR rate of 33%. Adverse events were observed in 32% of subjects, with mood symptoms being most common. There were no serious adverse events related to the study. Conclusion: We conclude that high dose oral dexamethasone is well tolerated by adults with idiopathic nephrotic syndrome and may have some efficacy.

scholarly journals Bcl-2 Protects Endothelial Cells against γ-Radiation via a Raf-MEK-ERK-Survivin Signaling Pathway That Is Independent of Cytochrome c Release

2007 ◽  
Vol 67 (3) ◽  
pp. 1193-1202 ◽  
Author(s):  
Pawan Kumar ◽  
Ila K. Coltas ◽  
Bhavna Kumar ◽  
Douglas B. Chepeha ◽  
Carol R. Bradford ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cytochrome C ◽  
Signaling Pathway ◽  
Cytochrome C Release ◽  
Γ Radiation

Aß(25-35) Peptide Induces Cell Death in PC12 Cells via Mitochondrial Damage and Cytochrome c Release

2005 ◽  
Vol 6 (1) ◽  
pp. 140-145 ◽  
Author(s):  
Cristiana Carelli Al . ◽  
Michela Pezzotti . ◽  
Daniele Mezzogori . ◽  
M. Elisabetta Clemen . ◽  
Francesco Misiti . ◽  
...  
Keyword(s):  
Cell Death ◽  
Cytochrome C ◽  
Pc12 Cells ◽  
Mitochondrial Damage ◽  
Cytochrome C Release

scholarly journals Resveratrol Attenuates Oxidized LDL-Evoked Lox-1 Signaling and Consequently Protects against Apoptotic Insults to Cerebrovascular Endothelial Cells

2010 ◽  
Vol 31 (3) ◽  
pp. 842-854 ◽  
Author(s):  
Huai-Chia Chang ◽  
Tyng-Guey Chen ◽  
Yu-Ting Tai ◽  
Ta-Liang Chen ◽  
Wen-Ta Chiu ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cytochrome C ◽  
Dna Fragmentation ◽  
Cell Apoptosis ◽  
Molecular Mechanisms ◽  
Cell Damage ◽  
Oxidized Ldl ◽  
Density Lipoprotein ◽  
Cytochrome C Release ◽  
Cerebrovascular Endothelial Cells

Cerebrovascular endothelial cells (CECs) are crucial components of the blood—brain barrier. Our previous study showed that oxidized low-density lipoprotein (oxLDL) induces apoptosis of CECs. This study was designed to further evaluate the effects of resveratrol on oxLDL-induced CEC insults and its possible molecular mechanisms. Resveratrol decreased the oxidation of LDL into oxLDL. Additionally, the oxLDL-caused oxidative stress and cell damage were attenuated by resveratrol. Exposure of CECs to oxLDL induced cell shrinkage, DNA fragmentation, and cell apoptosis, but resveratrol defended against such injuries. Application of Lox-1 small interference (si)RNA into CECs reduced the translation of this membrane receptor, and simultaneously increased resveratrol protection from oxLDL-induced cell apoptosis. By comparison, overexpression of Lox-1 attenuated resveratrol protection. Resveratrol inhibited oxLDL-induced Lox-1 mRNA and protein expressions. Both resveratrol and Lox-1 siRNA decreased oxLDL-enhanced translocation of proapoptotic Bcl-2-associated X protein (Bax) from the cytoplasm to mitochondria. Sequentially, oxLDL-induced alterations in the mitochondrial membrane potential, cytochrome c release, and activities of caspases-9, -3, and -6 were decreased by resveratrol. Pretreatment with Z-VEID-FMK (benzyloxycarbonyl-Leu-Glu-His-Asp-fluoromethyl ketone) synergistically promoted resveratrol's protection against DNA fragmentation and cell apoptosis. Therefore, this study shows that resveratrol can protect CECs from oxLDL-induced apoptotic insults via downregulating Lox-1-mediated activation of the Bax-mitochondria—cytochrome c—caspase protease pathway.

Mechanistic view for toxic effects of arsenic on isolated rat kidney and brain mitochondria

Biologia ◽  
2015 ◽  
Vol 70 (5) ◽  
Author(s):  
Shokoufeh Hassani ◽  
Hashem Yaghoubi ◽  
Roya Khosrokhavar ◽  
Iman Jafarian ◽  
Vida Mashayekhi ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cytochrome C ◽  
Outer Membrane ◽  
Brain Mitochondria ◽  
Male Rats ◽  
Arsenic Toxicity ◽  
Cytochrome C Release ◽  
Oxidation Stress ◽  
Complex Ii ◽  
Ros Formation

AbstractArsenic is one of the most important risk factors for human health and exhibits carcinogenicity in human. Emerging lines of research indicate that mitochondria are important target organelles for metals toxicity in living cells. In this study male rats were sacrificed and then kidney and brain mitochondria were isolated using ultracentrifugation method. Then, multi-parametric assays including reactive oxygen species (ROS) formation, complex II and IV activity, outer membrane integrity, ATP level and release of cytochrome c release evaluated to predict the biochemical pathways involved in arsenic toxicity. Our results showed that arsenic (25-200 μM) induced significant ROS formation rise and mitochondrial outer membrane damage in kidney and brain mitochondria, mitochondrial membrane potential collapse and mitochondrial ATP levels. Significant decrease in the complex II and IV activity in brain without any change in kidney mitochondria suggests the inevitable role of oxidative stress in mitochondrial permeability transition-mediated cytochrome c release. Therefore, it is supposed that mitochondrial oxidative stress and uncoupling of oxidative phosphorylation may play key roles in arsenic toxicity towards isolated kidney and brain mitochondria. Also, comparison of present study with previous investigations supposed that liver is more susceptible to arsenic exposure and induction of oxidation stress-like condition than kidney and brain tissues.

scholarly journals Ocular and extra-ocular complications following long-term steroid consumption in children with idiopathic nephrotic syndrome

2020 ◽  
Vol 11 (1) ◽  
pp. 7-10
Author(s):  
Gulshan Nigar Chaudhury ◽  
Tarannum Khandaker ◽  
Tahmina Ferdous ◽  
Rifat Asma Chowdhury ◽  
Shireen Afroz ◽  
...  
Keyword(s):  
Nephrotic Syndrome ◽  
Intraocular Pressure ◽  
Bone Pain ◽  
Idiopathic Nephrotic Syndrome ◽  
Epigastric Pain ◽  
Cross Sectional Study ◽  
High Dose ◽  
Ocular Complications ◽  
Cross Sectional ◽  
The Mean

Background: High dose of corticosteroids are required by patients with idiopathic nephrotic syndrome for long duration due to repeated relapse. The aims of this study were to analyze the ocular and extra-ocular complications of steroids on children with nephrotic syndrome. Methods: This cross-sectional study was conducted at Dhaka Shishu (Children) Hospital from September 2017 to September 2019. Children with nephrotic syndrome between 1-16 years of age, who received steroid for at least three months, participated in this study. Total 70 children were enrolled. Detailed history was taken from parents regarding initial episode, drug history, visual problem, headache, hypertension, weight gain, psychosis, bone pain, epigastric pain and excessive growth of body hair. Comprehensive opthalmologic assessment including visual acquity, intraocular pressure and cataract were performed. Detail of renal histopathology and treatment regimen in each patient was noted. Results: A total of 70 patients were included, 61.4% were male and 38.6% were female. Median age at the time of examination was 60 months (range 17 to 216 months). The mean duration of disease was 33.71±21.12 months. The mean cumulative steroid dose at the time of examination was 8485.91±7326.83 mg. Fourteen (20%) patients had posterior subcapsular cataract. Among 70 patients, we could do visual acuity for 25 (35.7%) patients, intraocular pressure (IOP) in mm for 51 (72.85%) patients. Among them raised IOP was found in five (9.8%) patients. In right eye mean IOP was 14.16±3.57 and for left eye 14.77±3.55 mm. Renal biopsy was performed in 15 (21.43%) cases. Among the biopsy report, it was found that mesangial proliferative glomerulonephritis (MPGN) in 8 (53.33%), focal segmental glomerulonephritis (FSGS) in 2 (13.33%) and minimal change in 5 (33.34%) cases. Among all cases, 12 (17.14%) patients developed Cushingoid facies, 3 (4.2%) patients had hypertrichosis, 3 (4.2%) patients had central obesity and 4 (5.7%) patients had buffalo hump but none had hypertension, diabetes mellitus, epigastric pain, bone pain or psychosis. Conclusion: Cataract formation was the most frequent ocular complication after large dose of oral corticosteroid therapy. The present study emphasizes the need for regular eye screening. Cushingoid facies was the most frequent among extra-ocular complications. Birdem Med J 2021; 11(1): 7-10

scholarly journals High-Dose Benzodiazepines Positively Modulate GABAA Receptors via a Flumazenil-Insensitive Mechanism

2021 ◽  
Vol 23 (1) ◽  
pp. 42
Author(s):  
Na Wang ◽  
Jingjing Lian ◽  
Yanqing Cao ◽  
Alai Muheyati ◽  
Shanshan Yuan ◽  
...  
Keyword(s):  
Gabaa Receptors ◽  
High Dose ◽  
High Concentration ◽  
Concentration Effects ◽  
In Vivo Evaluation ◽  
Chemical Structures ◽  
Loss Of Righting Reflex ◽  
High Concentrations

Benzodiazepines (BZDs) produce versatile pharmacological actions through positive modulation of GABAA receptors (GABAARs). A previous study has demonstrated that high concentrations of diazepam potentiate GABA currents on the α1β2γ2 and α1β2 GABAARs in a flumazenil-insensitive manner. In this study, the high-concentration effects of BZDs and their sensitivity to flumazenil were determined on synaptic (α1β2γ2, α2β2γ2, α5β2γ2) and extra-synaptic (α4β2δ) GABAARs using the voltage-clamp electrophysiology technique. The in vivo evaluation of flumazenil-insensitive BZD effects was conducted in mice via the loss of righting reflex (LORR) test. Diazepam induced biphasic potentiation on the α1β2γ2, α2β2γ2 and α5β2γ2 GABAARs, but did not affect the α4β2δ receptor. In contrast to the nanomolar component of potentiation, the second potentiation elicited by micromolar diazepam was insensitive to flumazenil. Midazolam, clonazepam, and lorazepam at 200 µM exhibited similar flumazenil-insensitive effects on the α1β2γ2, α2β2γ2 and α5β2γ2 receptors, whereas the potentiation induced by 200 µM zolpidem or triazolam was abolished by flumazenil. Both the GABAAR antagonist pentylenetetrazol and Fa173, a proposed transmembrane site antagonist, abolished the potentiation induced by 200 µM diazepam. Consistent with the in vitro results, flumazenil antagonized the zolpidem-induced LORR, but not that induced by diazepam or midazolam. Pentylenetetrazol and Fa173 antagonized the diazepam-induced LORR. These findings support the existence of non-classical BZD binding sites on certain GABAAR subtypes and indicate that the flumazenil-insensitive effects depend on the chemical structures of BZD ligands.

High-Dose Calcitriol Modulates the Expression and Activity of Tissue Factor and Thrombomodulin in Tumor Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 921-921
Author(s):  
Enriqueta Coll-Sangrona ◽  
Ali Amirkhosravi ◽  
Alshad S. Lalani ◽  
Liza Robles ◽  
Hina Desai ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Endothelial Cells ◽  
Tumor Cells ◽  
High Dose ◽  
Dependent Manner ◽  
High Concentrations ◽  
Dose Dependent ◽  
A549 Lung

Abstract Calcitriol, the hormonally-active metabolite of Vitamin D3, plays critical roles in calcium homeostasis, cell growth and differentiation, and immunoregulation. The anti-tumor activities of high-dose calcitriol have been demonstrated in a variety of preclinical models of solid tumors, leukemias and lymphomas. Recently, a new dose-intense formulation of calcitriol, termed DN-101 (Asentar™), was developed specifically for cancer therapy which allows for supraphysiological concentrations of calcitriol to be safely delivered in vivo to patients with cancer. In a recent Phase 2 clinical trial, DN-101 significantly increased overall survival and also reduced the incidence of thromboembolic events in men with androgen-independent prostate cancer receiving docetaxel-based chemotherapy. Based on previous observations we hypothesized that calcitriol’s anti-thrombotic effects in vivo may be due to the downregulation of Tissue Factor (TF) antigen and activity and/or upregulation of Thrombomodulin (TM). To test this hypothesis, we incubated A549 lung carcinoma, A375-C15 metastatic melanoma, THP-1 monocytic leukemia, and Eahy926 endothelial cells with increasing concentrations of calcitriol for 24 hrs. For TF induction, tumor cells were stimulated with TNFα for 5 hrs and activity was measured by a clotting assay and a thrombin generation assay (TGA). TM activity was measured by a chromogenic assay. TF and TM surface antigen were assessed by flow cytometry. Calcitriol prevented the induction of TF in TNFα-stimulated THP-1 cells in a dose-dependent manner (from 33% at 1 nM to 94% at 100 nM) as evidenced by a prolongation of plasma clotting time, a decrease in endogenous thrombin potential (ETP), and a reduction of surface TF antigen. In addition, the activity and surface expression of TM on THP-1 cells was increased significantly (40% and 3-fold respectively, P < 0.01) following 100 nM calcitriol treatment. Similarly, in TNFα-stimulated melanoma cells, calcitriol prevented the induction of TF activity (from 26% at 1 nM to 60% at 1 μM) and expression in a dose-dependent manner. High-dose calcitriol treatment also increased melanoma cell TM activity between 8% and 62%. In contrast, constitutively expressed TF activity and antigen were less affected by calcitriol in A549 lung carcinoma cells (12 to 28% reduction at concentrations between 1–100 nM) whilst TM activity and antigen were unaffected. In comparison to the tumor cells, calcitriol had no significant effect on TM or TF activity or antigen in TNFα-stimulated EAhy926 endothelial cells. In conclusion, we have demonstrated that high concentrations of calcitriol inhibit the induction of surface TF expression and upregulates TM in multiple tumor cell lines in vitro. The degree of the inhibition is proportional to the extent of TF induction by TNF-α. These in vitro results provide further support for the anticoagulant properties associated with high concentrations of calcitriol and may provide a rationale for understanding the lower incidence of thromboembolic complications observed in patients with metastatic prostate cancer treated with DN-101.

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