Anti-apoptotic Factor Research Articles

Val-Leu-Leu-Tyr (VLLY) Alleviates Ethanol-Induced Gastric Mucosal Cell Impairment by Improving Mitochondrial Fission.

Ethanolic gastric mucosal impairment is one of the most common disorders in the gastrointestinal system. In this study, we investigated the potential alleviating effects of sea cucumber peptides on Ges-1 impairment caused by ethanol and the associated mechanisms. The sea cucumber peptide VLLY could promote the proliferation and migration of healthy Ges-1 cells. After ethanol injury, VLLY peptide treatment could greatly promote the migration of Ges-1 cells, scavenge intracellular and mitochondrial ROS, reverse mitochondrial fission and F-actin depolymerization, and improve mitochondrial respiration. VLLY peptide restored mitochondrial dynamics by downregulating Drp1 and Fis1 and upregulating Mfn2 against excessive mitochondrial fission. In addition, the VLLY peptide maintained the mitochondrial membrane potential, ablated the leakage of cytochrome c to the cytoplasm, upregulated the expression of the antiapoptotic factor Bcl-XL, decreased the expression of the proapoptotic factors of Bax, BAD, and cleaved caspase-3, and finally blocked the mitochondria-related apoptotic pathway. These findings strongly suggested that sea cucumber peptides could promote proliferation and migration of healthy Ges-1 cells and reverse ethanol-induced excess mitochondrial fission and maintain mitochondrial homeostasis through the Fis1/Bax pathway, thereby improving ethanol-induced apoptosis. VLLY offers a new perspective for improving the ethanolic gastric mucosal epithelial cell injury.

Allium sativum Essential Oil Supplementation Reverses the Hepatic Inflammation, Genotoxicity and Apoptotic Effects in Swiss Albino Mice Intoxicated with the Lead Nitrate.

Prolonged lead (Pb) exposure impairs human health due to its interference with physiological and biochemical processes. Therefore, it is necessary to investigate natural therapeutics to alleviate Pb-induced intoxication. In the current investigation, essential oil extracted from the fresh bulbs of Allium sativum was considered as a natural remedy. Initially, in vitro antioxidant and anti-inflammatory activity of A. sativum essential oil (ASEO) were explored. The results reported that ASEO exhibits potent antioxidant and anti-inflammatory potential. Additionally, an in vivo study was conducted to elucidate its preventive role against Lead-nitrate (LN)-induced hepatic damage in Swiss albino mice. The experimental mice were allocated into six groups: Control, LN-intoxicated group (50mg/kg), LN + ASEO (50mg/kg), LN + ASEO (80mg/kg), LN + Silymarin (25mg/kg), and LN + vehicle oil control group. The entire duration of the study wasof 30days. From the results, it was determined that LN exposure elevated the Pb content in hepatic tissues which subsequently increased the serum biomarkers, inflammatory cytokines (NF-kB, TNF-α, IL-6) as well as apoptotic factors (caspase-3, BAX), all of which contribute to DNA damage. Meanwhile, it reduced anti-inflammatory (IFN-γ and IL-10) and anti-apoptotic factors (Bcl-2). Furthermore, Pb accumulation in hepatic tissues changed the histological architecture, which was linked to necrosis, central vein dilation, inflammatory cell infiltration and Kupffer cell activation. In contrast to this, ASEO administration decreased the Pb content, which in turn reduced the level of serum biomarkers, inflammatory and apoptotic factors. At the same time, it increased the anti-inflammatory and anti-apoptotic factors, thereby reduced DNA damage and restored the hepatic histology. In conclusion, exhaustive research is of the utmost demand to elucidate the precise defense mechanisms of ASEO against LN-induced hepatotoxicity.

Jiangqi Pingxiao formula regulates dendritic cell apoptosis in an autophagy-dependent manner through the AMPK/mTOR pathway in a murine model of OVA-induced asthma

Ethnopharmacological relevanceAllergic asthma is a recurring respiratory condition that typically manifests during childhood or adolescence. It is characterized by a dominant type II immune response triggered by the identification and capturing of inhaled allergens by dendritic cells (DCs). Jiangqi Pingxiao Formula (JQPXF), a prescription medicine used for the treatment of pediatric asthma, has been clinically proven to be both safe and effective. However, its mechanism of action in the treatment of asthma has not been fully been fully elucidated. Recent research suggests that several natural compounds have the potential to target dendritic cells (DCs) and alleviate ovalbumin (OVA)-induced asthma, which may also be found within JQPXF. Aim of the studyThis study aimed to elucidate the effect of JQPXF on OVA-induced asthma model and its molecular mechanism targeting DCs. Materials and methodsThe main constituents of JQPXF were analyzed by ultra performance liquid chromatography (UPLC). An asthma model was established by OVA. Hematoxylin-eosin staining and measurement of respiratory function was used to evaluate the treatment effect of JQPXF on asthmatic mice. Cytokine (IL-5, IL-13 and IgE) concentrations were determined by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was employed to evaluate inflammatory cell infiltration (T helper 2 cells and DCs) in vivo and DC survival in vivo and vitro. Western blot and immunofluorescence were used to verify the molecular mechanisms. ResultsThe results suggest that JQPXF can ameliorate pathological conditions and improve lung function in asthmatic mice, as well as the Th2 cells. Treatment with JQPXF significantly reduced the number of DCs and increased the number of Propidium iodide+ (PI) DCs. Furthermore, JQPXF upregulated protein levels of the pro-apoptotic factors Cleaved-caspase-3 and Bax, while downregulating the anti-apoptotic factor Bcl-2. Simultaneously, JQPXF increased autophagy levels by facilitating p62 degradation and promoting translation from LC3B I to LC3B II of DCs in vitro, as well as reducing the integrated optical density (IOD) of p62 within the CD11c-positive area in the lung. 3-Methyladenine (3-MA) was used to block autophagic flux and the apoptotic effect of JQPXF on DCs was abolished in vitro, with the number of DCs decreased by JQPXF being reversed in vivo. We further investigated the upstream key regulator of autophagy, the AMPK/mTOR pathway, and found that JQPXF increased AMPK phosphorylation while decreasing mTOR phosphorylation levels. Additionally, we employed Compound C (CC) as an AMPK inhibitor to inhibit this signaling pathway, and our findings revealed that both autophagic flux and apoptotic levels in DCs were abolished in vitro. ConclusionsIn summary, we have demonstrated that JQPXF could alleviate type II inflammation in an asthmatic model by promoting the apoptosis of DCs through an autophagy-dependent mechanism, achieved by regulating the AMPK/mTOR signaling pathway.

Quercetin alleviates zearalenone-induced apoptosis and necroptosis of porcine renal epithelial cells by inhibiting CaSR/CaMKII signaling pathway

Zearalenone (ZEA) is a mycotoxin that is highly contaminated in feed and can cause severe toxic effects on the kidneys and other organs of animals. Quercetin (QUE) is a plant-derived flavonoid with a variety of detoxification properties, but the mechanism by which QUE detoxifies the toxic effects induced by ZEA has not yet been fully elucidated. We treated porcine kidney cells (PK15) with 80 μM ZEA and/or 30 μM QUE. The results showed that ROS and MDA levels were increased, antioxidant system levels were down-regulated, anti-apoptotic factor expression levels were decreased, and apoptotic and necroptosis-related factors were up-regulated after ZAE exposure. In addition, the results of Ca2+ staining, mitochondrial membrane potential, and mitochondrial dynamics-related indicators showed that ZEA induced Ca2+ overload in PK15 cells and increased mitochondrial Ca2+ uptake (MCU expression increased). The accumulated ROS and free Ca2+ further aggravate mitochondrial damage and eventually lead to mitochondrial pathway apoptosis and necroptosis. Nevertheless, QUE targets CaSR to inhibit the CaSR/CaMKII pathway and regulate calcium homeostasis, thereby alleviating apoptosis and necroptosis mediated by mitochondrial dynamic disorder and dysfunction. The present study demonstrated the mechanism by which ZEA induces apoptosis and necroptosis in PK15 and the protective role of QUE in this process.

Sfrp1 as a Pivotal Paracrine Factor in the Trained Pericardial Stem Cells that Foster Reparative Activity.

Tissue damage often induces local inflammation that in turn dictates a series of subsequential responses, such as stem cell activation and growth, to maintain tissue homeostasis. The aim of the study is to testify the possibility of using inflammation-trained stem cells as optimal donor cells to augment the efficacy of cell therapy. The pericardial stem/stromal cells derived from the animals after myocardial infarction (MI-pSC) showed an enhanced myogenic potential and augmented reparative activity after transplantation in the injured hearts, as compared to the Sham-pSC. Bulk RNA-Seq analysis revealed significant upregulation of a panel of myogenic and trophic genes in the MI-pSC and, notably, Sfrp1 as an important anti-apoptotic factor induced robustly in the MI-pSC. Injection of the MI-pSC yielded measurable numbers of surviving cardiomyocytes (Tunel and Casp-3 negative) within the infarct area, but the effects were significantly diminished by siRNA-based silence of Sfrp1 gene in the pSC. Primed Sham-pSC with pericardial fluid from MI rats mimicked the upregulation of Sfrp1 and enhanced myogenic potential and reparative activity of pSC. Taken together, our results illustrated the inflammation-trained pSC favor a reparative activity through upregulation of Sfrp1 gene that confers anti-apoptotic activity in the injured cardiomyocytes. Therefore, the active form of stem cells may be used as a cardiac protective agent to boost therapeutical potential of stem cells.

ASXL1 mutations are associated with a response to alvocidib and 5-azacytidine combination in myelodysplastic neoplasms.

Inhibitors of anti-apoptotic BCL-2 family proteins in combination with chemotherapy and hypomethylating agents (HMA) are promising therapeutic approaches in acute myeloid leukemia (AML) and high-risk myelodysplastic syndromes (MDS). Alvocidib, a cyclin-dependent kinase 9 (CDK9) inhibitor and indirect transcriptional repressor of the anti-apoptotic factor MCL-1, has previously shown clinical activity in AML. Availability of biomarkers for response to the alvocidib + 5-azacytidine (5-AZA) could also extend the rationale of this treatment concept to high-risk MDS. In this study, we performed a comprehensive in vitro assessment of alvocidib and 5-AZA effects in N=45 high-risk MDS patients. Our data revealed additive cytotoxic effects of the combination treatment. Mutational profiling of MDS samples identified ASXL1 mutations as predictors of response. Further, increased response rates were associated with higher gene expression of the pro-apoptotic factor NOXA in ASXL1-mutated samples. The higher sensitivity of ASXL1 mutant cells to the combination treatment was confirmed in vivo in ASXL1Y588X transgenic mice. Overall, our study demonstrated augmented activity for the alvocidib + 5-AZA combination in higher-risk MDS and identified ASXL1 mutations as a biomarker of response for potential stratification studies.

Effects of Hyperventilation of Fine Particulate Matter (PM2.5) During a Single Exercise Bout on Inflammatory and Apoptotic Responses in the Mouse Lung

PURPOSE: The purpose of this study was to investigate the effects of hyperventilation in the presence of fine particulate matter (PM2.5) on inflammatory and apoptotic responses in the lungs of mice during a single bout of aerobic exercise.METHODS: The FVB/N mice were housed in a controlled SPF room and randomly divided into four groups: control (CON, n=7), fine particulate matter exposure (PM, n=7), PM and exercise (PME, n=7), and exercise only (EX, n=7). Acute PM exposure and/or aerobic treadmill exercise were undertaken according to the groups using a specially designed PM treadmill chamber.RESULTS: Protein expression and phosphorylation of p65 NF-kB in mouse lungs were significantly increased by acute exercise, but not by PM exposure. Basal protein levels and phosphorylation of MAPKs (p38, ERK, and JNK) were significantly increased in the PME group, but were lower in the PM and EX groups. Although the percentage of collagen fibers in lung tissue was not affected by PM exposure, the expression of pro- and antiapoptotic factors BAX, BID, and Bcl-2 was significantly elevated by inhalation of PM during exercise.CONCLUSIONS: In this study, the MAPKs family was activated by excessive PM inhalation during exercise and similar changes in apoptosis-related signaling were observed. This suggests that even a single bout of inhalation of fine particulate matter during exercise can affect factors associated with inflammation and cell death.

Apoptotic effects of mesenchymal stem cells’ conditioned medium on colorectal cancer cell lines

Multipotent Mesenchymal stem cells (MSCs) have vigorous immunomodulatory activity, apoptotic effects, and the capacity to migrate to inflammatory and tumor sites. This study focuses on the apoptotic effects of MSCs conditioned medium (CM) on colorectal cancer cell lines. MSCs were preconditioned with lipopolysaccharide (LPS) to induce apoptosis in colorectal cancer cells. The conditioned medium (LPS-CM) from the preconditioned cells was isolated and used to treat colorectal cancer cells (HT29 and SW48). The survival and proliferation of cancer cells were assessed using Trypan blue staining and MTT assay. The apoptosis rate was evaluated through flow cytometry analysis and caspase-3 activity. Additionally, Real-Time PCR was used to measure the mRNA level of apoptotic and anti-apoptotic factors, including bcl2, bax, and p53 genes. The results showed that LPS-CM significantly increased (p < 0.001) the percentage of apoptosis in the SW48 and HT29 cell lines. Caspase-3 activity significantly increased (p < 0.001) in these cell lines after treatment with LPS-CM. The mRNA level of bcl2 was significantly decreased (p < 0.001), while bax and p53 genes were significantly overexpressed (p < 0.001) in the LPS-CM treated cell lines. Notably, the mRNA level of bcl2 and bax genes was significantly altered at a higher concentration of LPS-CM. In conclusion, the conditioned medium from LPS-preconditioned MSCs can effectively induce apoptosis in colorectal cancer cells. This finding suggests that LPS-CM could be a potential strategy for inhibiting the proliferation and progression of colorectal cancer cells.

Homoharringtonine sensitizes pancreatic cancer to erlotinib by direct targeting and miRNA-130b-3p-mediated EphB4-JAK2-STAT3 axis.

Pancreatic cancer (PC) is a very lethal malignancy with a scarcity of treatment options. Although erlotinib- and gemcitabine-based treatments have been approved for PC, their effectiveness is limited. The present study is aimed at exploring the molecular and epigenetic mechanisms of anticancer activities of homoharringtonine (HHT) and its interaction with erlotinib to develop a potential therapeutic strategy for PC. The RT-qPCR, western blotting, immunofluorescence and expression-vectors and oligonucleotide transfection were employed to determine the expression characteristics of onco-factors. Anticancer activities were determined by MTT, colony forming, and flowcytometric analysis. Dual luciferase assay was conducted to confirm putative target of miR-130b-3p. In-vivo experiments were followed by immunohistochemical assay. The EphB4/JAK2/STAT3 pathway drives the growth and proliferation of PC through induction of prosurvival factors and cell cycle mediators. HHT directly and epigenetically via miR-130b-3p targets EphB4, leading to downregulation of JAK2/STAT3 pathway. The inactivation of STAT3 results in diminution of antiapoptotic factors and cell cycle mediators. HHT also enhances the anticancer activity of erlotinib. HHT demonstrates potential anticancer activities in PC by downregulating EphB4/JAK2/STAT3 signalling. HHT also produces synergistic effects with erlotinib.

Electroacupuncture Pretreatment Attenuates Learning Memory Impairment Induced by Repeated Propofol Exposure and Modulates Hippocampal Synaptic Plasticity in Rats.

Recurrent propofol anesthesia in the peak of neurodevelopment may lead to learning-memory decline. This study aimed to examine the efficacy of electroacupuncture pretreatment in ameliorating the aforementioned learning memory deficits and to explore its underlying mechanisms in a rat model of repeated propofol exposure. 10-day-old Sprague Dawley rats were randomly assigned to five groups: the control, fat emulsion, propofol, electroacupuncture pretreatment and electroacupuncture pretreatment combined with propofol groups. The electroacupuncture pretreatment involved three consecutive daily sessions, while propofol was received intraperitoneally once daily for five days. Following the modeling period, the rats' learning-memory performance was assessed using the New Novel Arm Y-maze, New Object Recognition, and Morris Water Maze. The Nissl staining method was used to observe the development of hippocampal neurons, while Golgi staining was employed to observe hippocampal synaptic development. The electroacupuncture pretreatment significantly attenuated the learning and memory impairment induced by recurring propofol exposure in rats. Additionally, it facilitated the development of hippocampal neurons and synaptic plasticity in the hippocampus. Immunofluorescence and Western Blot analyses were conducted to detect the expression of proteins related to apoptosis, learning memory, and synaptic plasticity. In the propofol group, the pro-apoptotic factors Caspase-3 and Bax was up-regulated, while the anti-apoptotic factor Bcl-2 was down-regulated, as compared to the blank group. Additionally, the phosphorylated cAMP-response element binding protein (pCREB), brain-derived neurotrophic factor (BDNF), synaptophysin, and growth associated protein-43 (GAP-43) was significantly decreased. In contrast, the electroacupuncture pretreatment combined with propofol group exhibited decreased the Caspase-3 and Bax and increased the Bcl-2, as compared to the propofol group, meanwhile, the pCREB, BDNF, Synaptophysin and GAP-43 was increased. Our findings indicate that electroacupuncture pretreatment can alleviate the learning and memory impairment induced by recurring propofol exposure in rats. This is achieved by enhancing hippocampal synaptic plasticity, activating the pCREB/BDNF pathway and inhibiting neuronal apoptosis.

Naringin is Protective in Paclitaxel-Induced Peripheral Neuropathy; A Multi-Biomarker Approach

Aim: Cancer is a disease that is on the rise worldwide. Paclitaxel (PTX) is one of the most common chemotherapeutic agents used in the treatment of many cancers. PTX causes toxic effects by increasing oxidative stress in tissues. Naringin is a powerful antioxidant found naturally in many plants, especially citrus fruits. The aim of this study was to determine the protective effects of NRG in PTX-induced sciatic nerve injury.&#x0D; Methods: Thirty-five male rats were randomly divided into five groups: control, PTX, NRG, PTX+NRG-50, PTX+NRG-100. PTX was administered i.p. for the first five days and NRG 50 or 100 mg/kg orally on days 6-14. Sciatic nerve tissues were harvested and analyzed for markers of oxidative stress, inflammation and apoptosis damage levels by biochemical methods.&#x0D; Results: PTX caused oxidative stress damage by increasing lipid peroxidation (MDA) and decreasing antioxidant capacity (SOD, CAT, GPx and GSH), inflammatory damage by increasing proinflammatory cytokine (NF-κB, TNF-α, IL-1β, SIRT1, TLR4, and NRF2) release, apoptotic damage by increasing apoptotic factor (Bax) and decreasing antiapoptotic factor (Bcl-2) in sciatic nerve tissue (p &lt; 0.05). NRG, on the other hand, reversed all these changes in sciatic nerve tissue and reduced PTX-induced oxidative stress damage, inflammatory damage and apoptotic damage (p &lt; 0.05). These effects were more effective at the 100 mg/kg dose of NRG than at the 50 mg/kg dose (p &lt; 0.05).&#x0D; Conclusions: In sciatic nerve tissue, PTX induced peripheral neuropathy with increased oxidative stress, inflammation and apoptotic damage. NRG showed a protective effect against PTX-induced peripheral neuropathy.

ApoE-modified liposomes encapsulating resveratrol and salidroside alleviate manifestations of Alzheimer’s disease in APP/PS-1 mice

Objective Alzheimer’s disease (AD) is a neurodegenerative disease that is associated with aging and is influenced by both genetic and environmental factors. Several studies and clinical trials have demonstrated that resveratrol (Res) and salidroside (Sal) are not only biologically safe but also influence AD biomarker trajectories. However, their clinical applications have been quite limited due to poor specificity, low solubility, and insufficient blood–brain barrier (BBB) penetration. Therefore, we developed a nano-drug delivery system in which Res and Sal were encapsulated in liposomes, which were surface-modified with ApoE (ApoE-Res/Sal-Lips) to compensate for these deficiencies. Method In this study, ApoE-Res/Sal-Lips were prepared using a standard thin-film hydration method for liposomes. Then, cellular uptake of the loaded liposomes was assessed in vitro using fluorescent staining assays. A BBB model was constructed to investigate the capacity of the liposomes to cross the BBB in vitro, and the ability of liposomes to target the brain was observed by in vivo imaging. In addition, the neuroprotective effects of the different liposome formulations in APP/PS-1 mice were evaluated by measuring the changes in levels of oxidative, anti-inflammatory, and anti-apoptotic factors in the mice brains. Results In vitro, ApoE-Res/Sal-Lips increased the uptake of Res and Sal by bEnd.3 and N2a cells, enhanced BBB penetration, and improved transport efficiency. In vivo, the ApoE-Res/Sal-Lips were found to alleviate AD pathological symptoms, reduce learning and memory impairments, and improve brain function. Conclusion ApoE-Res/Sal-Lips provide a new method for the treatment of AD.

Boric acid is associated with the suppression of apoptosis and endoplasmic reticulum stress in rat model of paracetamol -induced hepatotoxicity

Paracetamol (also known as acetaminophen or APAP) is an analgesic and antipyretic drug that is widely used all over the world. Boron is an element proven by many studies that it has indispensable effects on human health. In light of the information expressed about boron, we investigated whether there are any effects of boron on paracetamol-induced liver damage. We analyzed the 8-hydroxyguanosine (8-OHG), ALT, AST, glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and nitric oxide (NO) levels in serum or erythrocyte and liver tissue. At the same time, TNF-α, IL-1β, IL-18 levels were examined in liver tissues to determine the effect of boron on inflammation caused by paracetamol in serum and liver tissue. In this study to determine the effect of ER stress-mediated apoptosis, signal transducers that lead the cell to ER stress-mediated apoptosis, IRE1, ATF6 and PERK, caspase 1, 3, 8, 9 and 12, which play a role in the apoptotic process, antiapoptotic factors bcl2, bcl-xL mRNA gene expressions were determined and the effect level of boron on ER stress-mediated apoptosis was determined by immunohistochemical methods. This study demonstrate that boric acid protects against paracetamol-induced liver damage in association with the augmentation of biomarkers of oxidative and nitrosative stress, inflammation and the ER stress / apoptosis axis in rats.

Moringa oleifera Modulates MPTP-induced Mitochondrial Dysfunction in Parkinson’s Mouse Model: An in silico and in vivo Analysis

Background: Parkinson’s disease (PD) is a movement-affecting neurodegenerative condition with an unclear etiology. Recent research suggests targeting poly-(adenosine 5-diphosphate-ribose) polymerase 1 (PARP1) as a potential therapeutic approach for PD treatment. Purpose: The purpose of this study is to assess the effect of an ethanolic extract of Moringa oleifera leaves (MOE) on a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinsonian mouse model, with a specific focus on investigating its potential to mitigate the effects of α-synuclein toxicity, oxidative stress–induced hyper-activation of PARP1, and mitochondrial dysfunction associated with PD pathology. Additionally, this study also intends to investigate the alterations in neurobehavioral and biochemical parameters associated with PD pathology. Materials and Methods: An in silico docking study was conducted to investigate the phytochemicals found in M. oleifera (MO, drumstick plant) leaves as the potent inhibitors of PARP1. An in vivo (neurobehavioral, biochemical, and western blot) study was conducted to assess the neuroprotective effect of MOE on the MPTP-induced Parkinsonian mouse model. Results: The results of in silico study showed that the phytochemicals found in MO leaves could be a potent inhibitor of PAPR1. The in vivo study results showed that MOE significantly ameliorated MPTP-induced neurobehavioral and biochemical deficits. MPTP-induced mitochondrial enzyme-complex deficits were found to be restored in MOE-treated mice. Additionally, the result obtained in the western blot analysis showed that MOE significantly restored the levels of tyrosine hydroxylase in MPTP-intoxicated mice. MOE enhanced the expression of the anti-apoptotic factor (Bcl-2) and suppressed the expression of pro-apoptotic factors (Bax and caspase-3). Additionally, the enhanced levels of α-synuclein and PARP1 were significantly suppressed by MOE. Conclusion: Our findings suggest that MOE may possess pharmacological properties that inhibit neuronal damage in MPTP-intoxicated mice. Thus, MOE could be used as a therapeutic agent that can protect dopaminergic neurons from PARP1-induced neuronal damage.

Nitro-oleic acid ameliorates erectile dysfunction in a streptozotocin-induced rat model of diabetes by inhibiting oxidative stress and apoptosis and activating the NO/cGMP pathway.

The major vascular complications associated with diabetes make the management of diabetic mellitus erectile dysfunction (DMED) a challenging endeavor. Notable factors contributing to DMED include oxidative stress, nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway activation, and apoptosis, while nitro-oleic acid (NO 2 -OA) has been shown to be beneficial in treating these aspects of this condition. We, herein, investigated the effects and possible mechanisms of NO 2 -OA on erectile function as assessed in a streptozotocin-induced rat model of diabetes. Our results revealed that the erectile function of DMED rats was significantly impaired compared with that of the control group. However, in response to 4 weeks of NO 2 -OA treatment, there was an improvement in erectile function. The expression of oxidative stress-related indicators was significantly increased and the NO/cGMP pathway was impaired in the DMED group. The expression of proapoptotic factors was increased, while that of antiapoptotic factors was decreased in the DMED group. Moreover, the cell morphology in the cavernous tissue of the DMED group also changed adversely. NO 2 -OA treatment significantly reversed all these changes observed in the DMED group. In conclusion, NO 2 -OA treatment partially improved erectile function in DMED rats through mechanisms that included inhibition of oxidative stress, activation of the NO/cGMP pathway, and a reduction in apoptosis.

Krüppel-like factor 12 regulates aging ovarian granulosa cell apoptosis by repressing SPHK1 transcription and sphingosine-1-phosphate (S1P) production

Oxidative stress triggered by aging, radiation, or inflammation impairs ovarian function by inducing granulosa cell (GC) apoptosis. However, the mechanism inducing GC apoptosis has not been characterized. Here, we found that ovarian GCs from aging patients showed increased oxidative stress, enhanced reactive oxygen species activity, and significantly decreased expression of the known antiapoptotic factor sphingosine-1-phosphate/sphingosine kinase 1 (SPHK1) in GCs. Interestingly, the expression of Krüppel-like factor 12 (KLF12) was significantly increased in the ovarian GCs of aging patients. Furthermore, we determined that KLF12 was significantly upregulated in hydrogen peroxide-treated GCs and a 3-nitropropionic acid-induced invivo model of ovarian oxidative stress. This phenotype was further confirmed to result from inhibition of SPHK1 by KLF12. Interestingly, when endogenous KLF12 was knocked down, it rescued oxidative stress-induced apoptosis. Meanwhile, supplementation with SPHK1 partially reversed oxidative stress-induced apoptosis. However, this function was lost in SPHK1 with deletion of the binding region to the KLF12 promoter. SPHK1 reversed apoptosis caused by hydrogen peroxide-KLF12 overexpression, a result further confirmed in an invitro ovarian culture model and an invivo 3-nitropropionic acid-induced ovarian oxidative stress model. Overall, our study reveals that KLF12 is involved in regulating apoptosis induced by oxidative stress in aging ovarian GCs and that sphingosine-1-phosphate/SPHK1 can rescue GC apoptosis by interacting with KLF12 in negative feedback.

Unravelling the Collective Calcium Dynamics of Physiologically Aged Astrocytes under a Hypoxic State In Vitro.

Astrocytes serve many functions in the brain related to maintaining nerve tissue homeostasis and regulating neuronal function, including synaptic transmission. It is assumed that astrocytes are crucial players in determining the physiological or pathological outcome of the brain aging process and the development of neurodegenerative diseases. Therefore, studies on the peculiarities of astrocyte physiology and interastrocytic signaling during aging are of utmost importance. Calcium waves are one of the main mechanisms of signal transmission between astrocytes, and in the present study we investigated the features of calcium dynamics in primary cultures of murine cortical astrocytes in physiological aging and hypoxia modeling in vitro. Specifically, we focused on the assessment of calcium network dynamics and the restructuring of the functional network architecture in primary astrocytic cultures. Calcium imaging was performed on days 21 ("young" astrocyte group) and 150 ("old" astrocyte group) of cultures' development in vitro. While the number of active cells and frequency of calcium events were decreased, we observed a reduced degree of correlation in calcium dynamics between neighboring cells, which was accompanied by a reduced number of functionally connected cells with fewer and slower signaling events. At the same time, an increase in the mRNA expression of anti-apoptotic factor Bcl-2 and connexin 43 was observed in "old" astrocytic cultures, which can be considered as a compensatory response of cells with a decreased level of intercellular communication. A hypoxic episode aggravates the depression of the connectivity of calcium dynamics of "young" astrocytes rather than that of "old" ones.

Exploring the role and mechanism of Fuzi decoction in the treatment of osteoporosis by integrating network pharmacology and experimental verification

BackgroundFuzi decoction (FZD), a traditional Chinese medicine formula, was used to treat musculoskeletal diseases by warming channels, strengthening yang and dispelling pathogenic cold and dampness. In clinical practice, FZD has been used to treat rheumatoid arthritis and osteoarthritis. It alleviated osteoarticular disorders through ameliorating the degradation of cartilage and improving meniscal damage in osteoarthritis, while its roles and mechanisms in the treatment of bone loss diseases remain unclear. This study aims to investigate the underlying mechanisms of FZD in treating osteoporosis using an integrative method of network pharmacology and experimental study.MethodsIn this study, network pharmacology was used to predict the core targets and potential pathways of the bioactive ingredients of FZD to attenuate osteoporosis. Molecular docking was performed to evaluate the interactions between core compounds and key targets. In addition, both cell and animal experiments were carried out to validate the role and potential mechanism in treating osteoporosis.ResultsIn the present study, data revealed that kaempferol, beta-sitosterol, stigmasterol, fumarine, and (+)-catechin may be the primary bioactive ingredients of FZD in the treatment of osteoporosis, which were closely associated with the osteoporosis-related targets. And the KEGG results indicated that the NF-κB pathway was closely associated with the function of FZD in treating osteoporosis. In addition, in vivo demonstrated that FZD ameliorated osteoporosis. In vitro experiments showed that the pro-apoptotic factors indicators including CASP3 and BAX were decreased by FZD and the anti-apoptotic factor BCL2 was increased by FZD. In addition, FZD significantly suppressed the osteoclast differentiation in culture and the expression levels of osteoclast-related genes including TRAF6, CTSK, and MMP9. And the NF-κB pathway was confirmed, via in vitro experiment, to be involved in osteoclast differentiation.ConclusionsThis study demonstrated that FZD played a pivotal role in suppressing the osteoclast differentiation via regulating the NF-κB pathway, indicating that FZD could be a promising antiosteoporosis drug and deserve further investigation.

Expression of NF-κB, Bcl-2 and Caspase-8 apoptosis markers in endometrium in external genital endometriosis and chronic endometritis: A retrospective study

Aim. To study the regulation of apoptosis in the endometrium of women with chronic endometritis (CE) and external genital endometriosis (EGE) based on the study of the expression of its regulators such as transcription factor (NF-), Bcl-2 and Caspase-8.&#x0D; Materials and methods. The study evaluated the expression of apoptosis factors such as Caspase-8, NF-B transcription factor, and Bcl-2, in patients with chronic endometritis (CE, n=23), external genital endometriosis (EGE, n=20), and intact endometrium (control group; n=7). Statistically significant differences were found for all test markers (p0.05).&#x0D; Results. The increased expression of apoptosis factors is probably related to high endometrial pro-inflammatory activity in both CE and EGE. For EGE, this effect is more pronounced at the transcription level in the form of an increase in the expression of the transcription factor NF-B in both glandular and stromal endometrial cells. It is probably due to the systemic nature of the chronic inflammatory process. For CE, despite the antiapoptotic activity of Bcl-2, an increase in Caspase-8 expression in the endometrium may also indicate a predominance of innate immunity activation.&#x0D; Conclusion. The imbalance of pro- and antiapoptotic factors can be an inducer of both cytotoxic and cytoprotective mechanisms of EGE and CE. However, it requires further research.

DSC2 suppresses the growth of gastric cancer through the inhibition of nuclear translocation of γ-catenin and PTEN/PI3K/AKT signaling pathway.

Globally, gastric cancer (GC) is still a major leading cause of cancer-associated deaths. Downregulated desmocollin2 (DSC2) is considered to be closely related to tumor progression. However, the underlying mechanisms of DSC2 in GC progression require further exploration. We initially constructed different GC cells based on DSC2 contents, established the mouse tumor xenografts, and subsequently performed clonal formation, MTT, Caspase-3 activity, and sperm DNA fragmentation assays to detect the functions of DSC2 in GC growth. Subsequently, we performed western blot, Co-IP, and immunofluorescence assays to investigate the underlying mechanisms through pretreatment with PI3K inhibitor, LY294002, and its activator, recombinant human insulin-like growth factor I (IGF1). DSC2 could significantly inhibit the viability of GC cells at both in vitro and in vivo levels. The underlying mechanism may be that DSC2 binds the γ-catenin to decrease its nuclear level, thereby downregulating the anti-apoptotic factor BCL-2 expression and upregulating the pro-apoptotic factor P53 expression, which adjusts the PTEN/PI3K/AKT signaling pathway to promote the cancer cell apoptosis. Our finding suggests that DSC2 might be a potential therapeutic target for the treatment of cancers, most especially GC.