Apoptotic Factors Research Articles

The Effect of a Period of Resistance Training on Some Apoptotic Markers in Kidney Tissue of Diabetic Male Rats with Morphine Withdrawal Syndrome

Background and Objective: The objective of this study was to investigate the effect of resistance training on some apoptotic markers of kidney tissue with morphine withdrawal syndrome in diabetic rats. Materials and Methods: In this experimental study, 32 male Wistar rats were randomly divided into 4 groups of 8, including: diabetes, morphine diabetes, diabetes + resistance training, diabetes morphine + resistance training. After implementing the diabetes induction protocol, the addicted groups received morphine orally for 21 days, and then the training groups participated in the resistance training protocol for 8 weeks. Then all the mice were killed and dissected and their kidney tissue was removed. ELISA kits were used to evaluate apoptotic factors. Results: Cytochrome-C variable results showed a significant decrease in the morphine diabetes (P=0.002) and diabetes + resistance training (P=0.001) and morphine diabetes + resistance training (P=0.001) groups compared to the diabetes group. The results of the caspase-8 variable also showed that the diabetes + resistance training group had a significant decrease compared to the diabetes group (P=0.004) and the morphine diabetes group (P=0.012). Also, the reduction of caspase 3 variable results in the morphine diabetes group (P=0.001), diabetes + resistance training (P=0.001) and morphine diabetes + resistance training (P=0.002) was significant compared to the diabetes group. Conclusion: Resistance training has reduced apoptotic indices in kidney tissue and can be a suitable treatment strategy for diabetes and withdrawal syndrome diabetes.

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.

Cichoric acid improves isoproterenol-induced myocardial fibrosis via inhibition of HK1/NLRP3 inflammasome-mediated signaling pathways by reducing oxidative stress, inflammation, and apoptosis.

Cichoric acid (CA), a natural phenolic compound found in many plants, has been reported to have antioxidant, anti-inflammatory, hypoglycemic, and other effects. The aim of this study was to determine the potential role and underlying mechanisms of CA in isoproterenol (ISO)-induced myocardial fibrosis (MF). The MF model was induced by subcutaneous ISO injection in mice. Blood and heart tissue were collected for examination. Hematoxylin and eosin staining and Masson's trichrome staining were used to evaluate the histopathological changes and collagen deposition. The production of reactive oxygen species markers was observed by fluorescence microscopy, the degree of cardiomyocyte microstructure injury was observed by transmission electron microscope, and oxidative stress factors were detected by kit method, and the effect of CA on inflammatory factors was detected by ELISA. The expression levels of collagen proteins and signaling pathways were further investigated by western blotting. The results showed that CA inhibited the expression of ISO-induced proinflammatory factors (TNF-α, IL-1β, and IL-18) and proteins (HK1, NLRP3, caspase-1, cleaved-caspase-1, and ASC), and regulated the expression of apoptotic factors (caspase-3, cleaved-caspase-3, Bax, and Bcl-2). The results indicated that CA may regulate the HK1/NLRP3 inflammasome pathway by inhibiting HK1 expression and play a protective role in MF.

Every-other day (EOD) feeding regime decreases oxidative stress and inflammatory cascade in mouse liver: The immunohistochemical study

IntroductionPositive effects of calorie restrictions (CR) and EOD include decreased body weight, prolonged life span, but also changes in metabolism of the liver. In present paper our aim was to examine antioxidative enzymes: Catalase (CAT) and Manganese superoxidative dismutase (MnSOD, SOD2) and Glutathione peroxidase 4 (Gpx-4) in connection to caspase-3 and inflammatory mediators (IL-1β and TNF- α) in EOD liver tissue in comparison to control mice. MethodsAfter 9 months of EOD treatment male mouse liver tissue was harvested and prepared for analysis. Protein semi-quantitative estimation and cellular immunolocalization was performed for CAT, SOD2, Gpx-4, caspase-3, IL-1β and TNF- α in liver tissue of mice fed every-other day in comparison to control (AL fed) animals. ResultsAfter prolonged EOD feeding in mice we observed decreased level of SOD2 and Gpx-4, decreased caspase-3, IL-1β and TNF-α expression in liver tissue on protein level measured by semi-quantitative DAB staining intensity. ConclusionFor the first time we showed immunolocalization of major antioxidative enzymes (CAT, SOD2, Gpx-4) in liver tissue after DR. Decrease of two major antioxidant enzymes combined with decrease of apoptotic marker and inflammatory factors indicate decrease in oxidative stress as the result of fast in EOD feeding regime.

The mechanism of acrolein exposure inhibited the release of neutrophil extracellular traps: By reducing respiratory burst and Raf/MEK/ERK pathway and promote cell apoptosis

Acrolein (AC) is a highly toxic volatile substance in the environment, and studies have found that excessive AC had a toxic effect on the immune system. Neutrophils are the first line of defense against pathogen invasion. The release of neutrophil extracellular traps (NETs) is a protective mechanism for neutrophils, and its release is affected by environmental pollutants. However, the effect of AC on NETs release and its mechanism remains unclear. In this study, chicken peripheral blood neutrophils were pretreated with 20 μM AC and treated with 5 μM Phorbol 12-myristate 13-acetate (PMA) to stimulate the release of NETs. The results showed that AC exposure significantly inhibited the release of NETs induced by PMA, respiratory burst, and the expression levels of phospho-rapidly accelerated fibrosarcoma (p-Raf), phospho-mitogen-activated extracellular signal-regulated kinase (p-MEK) and phospho-extracellular regulated protein kinases (p-ERK). In addition, AC exposure significantly inhibited the expression of B-cell lymphoma-2 (Bcl-2) and promoted the expression of apoptotic factors Bcl2-Associated X (Bax), cytochrome c (Cyt C), cysteinyl aspartate specific proteinase 9 (Casp 9) and cysteinyl aspartate specific proteinase 3 (Casp 3). Further inhibition of neutrophil apoptosis significantly improved the release of NETs. The above results indicated that AC exposure led to a decrease in the formation of NETs, which is caused by excessive AC-induced neutrophil apoptosis. Our study clarified the immune toxicity mechanism of AC on chickens, which is of great significance and reference value for protecting the ecological environment and poultry health.

The Effect of Vitamin E on Intestinal Epithelial Cells in Broiler Subjected to Heat Stress

Background: Heat stress leads to great economic losses in poultry industry because of increasing climate change globally. There are several nutrients which are generally used as to combat against heat stress. Vitamin E is one of the feed additives used to prevent losses due to heat stress. This study investigated the protective effect of vitamin E against cellular damage that may occur in intestinal epithelial cells against the heat stress. There is no information on the response of vitamin E to cellular damage in intestinal epithelial cells in broilers under heat stress. Methods: In the current study, damage and the response to damage were tried to be revealed by considering both apoptotic factors, necrosis and inflammation markers. For this purpose, a total of 30 broiler chicks were divided into three replicates and analyzed. The control group was kept at 24°C, while the heat stress group was subjected to heat stress at 35°C (5 hours/day). Finally, 300 mg/kg vitamin E (α-tocopherol acetate) was supplemented daily to the heat stress+vitamin E group while heat stress was applied at 35°C (5 h/day). Result: In the study, immuno-stainings were performed with caspase-9 to detect the intrinsic pathway of apoptosis, caspase-8 and TNF-α to detect the extrinsic pathway of apoptosis and IL-6 primary antibodies to detect the pro-inflammatory immune response. Consequently, it was determined in the study that caspase-9 activity, which was positive only in epithelial cells in all groups, increased significantly in heat stress, but vitamin E decreased this activity by 75%. However, caspase-8 activity increased with heat stress in both epithelial and connective tissue and vitamin E had a protective effect. It was observed that TNF-α activity increased with heat stress in all three parts of the small intestine, especially in the duodenum. However, this activity in the group with vitamin E was closer to the control group. IL-6 activity, an indicator of pro-inflammatory response, caused no difference between the groups. As a result, the study indicates that apoptosis may occur even if there is no acute systemic inflammation in the intestinal epithelial cells of living organisms exposed to heat stress. Additionally, this study reveals that vitamin E may reduce the effect of apoptosis in the intestines against heat stress.

The delaying effect of Qingxin Zishen decoction on ovarian aging: Examining regulation of ovarian mitochondria apoptosis in aged rats

Ovarian aging is a significant challenge in gynecology, and there is currently no effective treatment for it. However, the medicinal agent Qingxin Zishen decoction (QZD) has shown potential in the treatment of ovarian dysfunction. The present study aimed to evaluate the mitochondrial apoptotic mechanism of delayed ovarian aging in QZD in aging rats. The healthy female Sprague-Dawley (SD) rats (n = 40, 350 ± 20 g) were randomly assigned to different dosage groups and 4-month-old SD rats (n = 10) were assigned to the control group. QZD groups were treated with QZD for four weeks, and ovarian tissues were extracted for mRNA and protein assays to examine the role of the apoptotic pathway in QZD. The results showed that QZD treatment for four weeks significantly increased the mRNA and protein expressions of the anti-apoptotic gene B-cell lymphoma 2 (Bcl-2) and Bcl-2/Bax ratio, as well as downregulated the pro-apoptotic genes Bax, caspase-3, and caspase-9. Moreover, QZD treatment effectively reduced the expression of cytochrome C (cyto-C) and apoptotic protease-activating factor 1 (Apaf-1), both of which are components of the intrinsic apoptotic pathway. These changes exhibited a dose-response manner. The findings suggest that QZD might have therapeutic potential in delaying ovarian mitochondrial function decline and in preventing and treating ovarian aging-related diseases by downregulating and upregulating the pro-apoptotic (Bax, caspase-3, caspase-9, cyto-C, Apaf-1) and anti-apoptotic (Bcl-2 and Bcl-2/Bax ratio) genes, respectively.

Alteration of pulmonary mitochondrial oxidation status, inflammation, energy metabolizing enzymes, oncogenic and apoptotic markers in mice model of diisononyl phthalate-induced asthma

Primary plasticizer used in polyvinyl chloride is diisononyl phthalate (DiNP) and exposure to DiNP has been associated with the development of asthma and allergies. In the current study, how DiNP alters pulmonary antioxidant status, inflammation, energy metabolizing enzymes, oncogenic and apoptotic markers in DiNP-induced asthmatic mice was examined. Male BALB/c mice (n=20, 20-30 g) were divided into 2 groups of 10 mice each: group 1 (control) received saline (0.2ml/kg) orally for 23 days, and group 2 (DiNP) received 50 mg/kg DiNP (Intraperitoneal and intranasal) once per day. After the last administration, mice were sacrificed, lungs were removed and used for biochemical and histopathological analysis. DiNP treated mice experienced alterations in their lung histoarchitecture, levels of oncogenic and apoptotic factors, glycolytic, tricarboxylic acid cycle (TCA), and electron transport chain enzymes (ETC), antioxidant status, and inflammatory biomarkers. DiNP decreased the lungs levels of reduced glutathione and ascorbic acid, and the activities of superoxide dismutase, catalase, and glutathione-s-transferase. In the lungs of DiNP-treated mice compared to the control group, malondialdehyde and inflammatory biomarkers (nitric oxide and myeloperoxidase) were significantly greater (p&lt;0.05). Furthermore, the activities of glycolytic enzymes hexokinase, aldolase, lactate dehydrogenase were downregulated with a concomitant increase in NADase (77%). TCA enzymes and ETC enzymes were significantly reduced as well. CAS-3, p53, Bax, c-MYC, K-Ras increased by 65%, 51%, 70%, 59% and 82% respectively while BCL-2 decreased by 74%. Histopathological analysis revealed distortion of the airway structure characterized by inflammatory cell infiltration, oedema, hemorrhage, and constricted alveoli space. Exposure to DiNP caused oxidative stress which promotes lung inflammation via depletion of antioxidants, pulmonary energy transduction enzymes, levels of oncogenic and apoptotic factors were impaired as well, suggesting that the lungs may not be able to perform its morphological and physiological functions effectively.

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.

Phenylalanine Plays Important Roles in Regulating the Capacity of Intestinal Immunity, Antioxidants and Apoptosis in Largemouth Bass (Micropterus salmoides).

This experiment was planned to explore the role of dietary phenylalanine levels in intestinal immunity, antioxidant activity and apoptosis in largemouth bass (Micropterus salmoides). Six iso-nitrogen and iso-energy diets with phenylalanine levels of 1.45% (DPHE1), 1.69% (DPHE2), 1.98% (DPHE3), 2.21% (DPHE4), 2.48% (DPHE5) and 2.76% (DPHE6) were designed. Juvenile largemouth bass were fed the experimental diet for 8 weeks. In this study, the DPHE5 group increased the expression of intestinal antioxidant genes in largemouth bass (p < 0.05), and the increase of antioxidant enzyme activities and content of related substances was most concentrated in the DPHE3 and DPHE4 groups (p < 0.05). The results of plasma biochemistry were similar to that of enzyme activity. The expression of genes related to the TOR signalling pathway mainly increased significantly in the DPHE5 group (p < 0.05). Similarly, the expression of inflammatory factors, as well as apoptotic factors, also showed significant increases in the DPHE5 group (p < 0.05). In conclusion, unbalanced phenylalanine in the diet could lead to a decrease in intestinal immune and antioxidant capacity and also cause a decline in the aggravation of intestinal cell apoptosis.

Complement C3 Reduces Apoptosis via Interaction with the Intrinsic Apoptotic Pathway.

Myocardial ischemia/reperfusion (I/R) elicits an acute inflammatory response involving complement factors. Recently, we reported that myocardial necrosis was decreased in complement C3-/- mice after heart I/R. The current study used the same heart model to test the effect of C3 on myocardial apoptosis and investigated if C3 regulation of apoptosis occurred in human cardiomyocytes. Comparative proteomics analyses found that cytochrome c was present in the myocardial C3 complex of WT mice following I/R. Incubation of exogenous human C3 reduced apoptosis in a cell culture system of human cardiomyocytes that did not inherently express C3. In addition, human C3 inhibited the intrinsic apoptosis pathway in a cell-free apoptosis system. Finally, human pro-C3 was found to bind with an apoptotic factor, pro-caspase 3, in a cell-free system. Thus, we present firsthand evidence showing that C3 readily reduces myocardial apoptosis via interaction with the intrinsic apoptotic pathway.

Arsenic and polystyrene-nano plastics co-exposure induced testicular toxicity: Triggers oxidative stress and promotes apoptosis and inflammation in mice.

Co-existing of polystyrene-nano plastics (PSNPs) and arsenic (As) in the environment caused a horrendous risk to human health. However, the potential mechanism of PSNPs and As combination induced testicular toxicity in mammals has not been elucidated. Therefore, we first explore the testicular toxicity and the potential mechanism in male Kunming mice exposed to As or/and PSNPs. Results revealed that compared to the As or PSNPs group, the combined group showed more significant testicular toxicity. Specifically, As and PSNPs combination induced irregular spermatozoa array and blood-testis barrier disruption. Simultaneously, As and PSNPs co-exposure also exacerbated oxidative stress, including increasing the MDA content, and down-regulating expression of Nrf-2, HO-1, SOD-1, and Trx. PSNPs and As combination also triggered testicular apoptosis, containing changes in apoptotic factors (P53, Bax, Bcl-2, Cytc, Caspase-8, Caspase-9, and Caspase-3). Furthermore, co-exposed to As and PSNPs aggravated inflammatory damage characterized by targeted phosphorylation of NF-κB and degradation of I-κB. In summary, our results strongly confirmed As + PSNPs co-exposure induced the synergistic toxicity of testis through excessive oxidative stress, apoptosis, and inflammation, which could offer a new sight into the mechanism of environmental pollutants co-exposure induced male reproductive toxicity.

Apoptin and apoptotic protease-activating factor 1 plasmid-assisted multi-functional nanoparticles in hepatocellular carcinoma therapy

Cancer drugs usually have side effects in chemotherapy. Apoptin, a protein recognized by its good therapeutical effect on tumors and innocuous to body, is employed to treat hepatocellular carcinoma (HCC). As our previous data shown, the efficiency of apoptin protein might be limited by the protein of apaf-1. Therefore, we designed the multi-functional nanoparticles (MFNPs) encapsulating apoptin and apaf-1 plasmids by layer-by layer assembly. The NPs could release drugs into tumor site specifically and had good compatibility to normal cells and tissues. The groups of biotin, ε-polylysine, and nuclear localization signal in MFNPs conferred NPs the capabilities to enter cancer cells specifically, escape lysosome and enter the nucleus, respectively. In vitro inhibition experiment and in vivo anti-tumor therapy confirmed MFNPs as an excellent carrier to treat HCC. In addition, the dual-drug system was superior to any of the single-drug system. The mechanism analysis proved that supplement of the protein of apaf-1 might enhance apoptosome formation, causing the increase of therapeutical efficacy.

High affinity of protocatechuic acid to human serum albumin and modulatory effects against oxidative stress and inflammation in alveolar epithelial cells: Modulation of pulmonary fibrosis

Protocatechuic acid (PCA), C7H6O4, has been shown to possess potential antioxidant properties. But its interaction with the main plasma carrier protein, human serum albumin (HSA), as well as its antioxidant mechanism remains largely unknown. It has been shown that induced pulmonary fibrosis can be modulated through mitigating epithelial apoptosis mediated by the prohibition of oxidative stress. Therefore, in this study, the interaction of PCA and HSA was explored by spectroscopy, calorimetry (DSC), as well as molecular docking studies. Also, the protective effects of PCA against lipopolysaccharide (LPS)-induced cytotoxicity and oxidative stress were evaluated by MTT, ROS, ELISA, real-time PCR, and western blot assays. It was shown that under the interaction of HSA with PCA a spontaneous interaction occurs with the contribution of hydrophobic forces, which results in the formation of a stable complex. Cellular assays showed that PCA reduced LPS-induced cytotoxicity in human type II alveolar epithelial cells (AECs) through mitigation of ROS production, release and gene expression of TNf-α and IL-1β as pro-inflammatory mediators, and caspase-3 gene and protein as an apoptotic factor. Also, it was shown that PCA can reduce the expression of NF-κB at the protein level, indicating a possible inhibition of pulmonary fibrosis via regulating the NF-κB signaling pathway. In conclusion, PCA could be a promising therapeutic agent for the control of oxidative stress in AECs which is an important factor in redox modulatory therapy, while its pharmacodynamics can be modulated by interaction with HSA.

Exercise improves cardiac function and attenuates myocardial inflammation and apoptosis by regulating APJ/STAT3 in mice with stroke

Stroke can induce cardiac dysfunction without a primary cardiac disease. Exercise can promote the overall rehabilitation of stroke patients and be beneficial for all kinds of heart diseases. However, the mechanisms underlying the protective effects of exercise in stroke-induced cardiac dysfunction are poorly understood. Hence, we aimed to distinguish the different effects of acute and long-term exercise and further study the mechanism of protection against cardiomyopathy caused by stroke. Mice underwent a single acute session or long-term exercise for 30 days, followed by middle cerebral artery occlusion surgery. The expression of apoptosis-related proteins and proinflammatory factors in the heart was evaluated. Then, overexpression of apelin peptide jejunum (APJ) transfected adeno-associated virus type 9 (AAV9) and inhibition of signal transducer and activator of transcription 3 (STAT3) by Stattic were used in stroke mice or hypoxic cardiomyocytes. ML221 were used to inhibit APJ activity in exercise mouse. Thereafter, changes in apoptotic and proinflammatory factors were evaluated. The results demonstrated that chronic exercise prevented myocardial inflammation, apoptosis and cardiac dysfunction after stroke. However, acute exercise did not have similar effects. Exercise maintained the levels of APJ expression and decreased phosphorylated-STAT3 (p-STAT3) activation to protect cardiomyocytes. Moreover, APJ overexpression promoted cardiomyocyte survival and reduced p-STAT3 levels. STAT3 inhibition also reduced apoptosis and proinflammatory factors in mice hearts. Conversely, the protective effect of exercise was eliminated by APJ inhibition. This study showed that exercise can maintain APJ expression and inhibit p-STAT3, thus, conferring protection against myocardial inflammation and apoptosis induced by stroke.

Apoptotic Factors Are Evolutionarily Conserved Since Mitochondrial Domestication.

The mechanisms initiating apoptotic programmed cell death in diverse eukaryotes are very similar. Basically, the mitochondrial permeability transition activates apoptotic proteases, DNases, and flavoproteins such as apoptosis-inducing factors (AIFs). According to the hypothesis of the endosymbiotic origin of apoptosis, these mechanisms evolved during mitochondrial domestication. Various phylogenetic analyses, including ours, have suggested that apoptotic factors were eubacterial protomitochondrial toxins used for killing protoeukaryotic hosts. Here, we tested whether the function of yeast Saccharomyces cerevisiae apoptotic proteases (metacaspases Mca1 and Nma111), DNase Nuc1, and flavoprotein Ndi1 can be substituted with orthologs from remotely related eukaryotes such as plants, protists, and eubacteria. We found that orthologs of remotely related eukaryotic and even eubacterial proteins can initiate apoptosis in yeast when triggered by chemical stresses. This observation suggests that apoptotic mechanisms have been maintained since mitochondrial domestication, which occurred approximately 1,800 Mya. Additionally, it supports the hypothesis that some of these apoptotic factors could be modified eubacterial toxins.

Dose-dependent neuroprotective effects of adipose-derived mesenchymal stem cells on amyloid β-induced Alzheimer's disease in rats

AimMesenchymal stem cells (MSCs) have emerged as an intriguing candidate in cell therapy for treating neurodegenerative diseases, including Alzheimer's disease (AD). To achieve the maximum efficiency of cell therapy, determining the optimal dose of MSCs is essential. This study was conducted to assess the dose-dependent therapeutic response of MSCs against pathological and behavioral AD-associated alterations. MethodsAβ1-42 was injected intrahippocampally to establish an AD rat model. The MWM test was utilized to evaluate the animal's behavioral functions after receiving low and high doses of MSCs in the hippocampus region. ELISA and RT-qPCR were also employed to assess the concentration of markers related to antioxidant activity and inflammation and the gene expression related to apoptosis in the hippocampus region, respectively. ResultsLow-dose MSC transplantation by increasing the concentrations of the antioxidant GSH, the anti-inflammatory cytokine IL-10, as well as by lowering the concentrations of TNF-α, and the expression levels of apoptotic factors (Bax and caspase 3), exerted a neuroprotective effect in the hippocampus of AD rats and relatively ameliorated spatial learning and memory impairments. However, increasing the dose of MSCs decreased the therapeutic benefits of these cells and had no significant effect on the recovery of behavioral disorders. ConclusionOur findings reveal the dose-dependent neuroprotective effect of MSCs in AD. The therapeutic response of MSCs to ameliorate the pathological and behavioral alterations associated with AD is attenuated when the dosage of MSCs is increased.

PROTECTIVE EFFECT OF ANTI-EPILEPTIC AGENT Q808 ON NEURONAL DAMAGE IN A RAT MODEL OF TEMPORAL LOBE EPILEPSY

The detailed effect of Q808 (6-(4-chlorophenoxy)-tetrazolo[5,1-a]phthalazine) which has been demonstrated to have anti-epileptic activity, on the protection from neuronal damage in temporal lobe epilepsy (TLE) still remains unclear. The purpose of this study was to evaluate the protective effects of Q808 on neuronal damage by exploring its mechanism of hippocampal neuronal apoptosis in a rat model of TLE. Fifty rats were divided equally into five groups. The normal group [Formula: see text] served as the normal non-seizure and untreated group. To induce the TLE model, pilocarpine (20[Formula: see text]mg/kg) was intraperitoneally administered to the other 40 rats; 33 rats successfully developed TLE status. Subsequently, the TLE group [Formula: see text], VPA group [Formula: see text], Q808-L group [Formula: see text], and Q808-H group [Formula: see text] were, respectively, treated with 5[Formula: see text]ml normal saline, 1[Formula: see text]ml/kg valproate, 0.5[Formula: see text]ml/kg Q808, and 1[Formula: see text]ml/kg Q808 solution once daily for 28 days. The Morris water maze test was performed to evaluate the effects of Q808 on cognitive impairment in the TLE model. Hematoxylin-eosin (HE) staining was performed to determine histopathological changes in the hippocampus, immunohistochemistry was performed to determine the expression of caspase-3 in the hippocampus, and western blotting was performed to determine the expression levels of apoptosis-related proteins, including Apaf-1, caspase-9, and caspase-3. Q808 decreased the expression levels of the apoptosis-related proteins in the TLE hippocampal tissue, and higher doses are more effective. Besides, Q808 did not impair spatial cognition and memory. Furthermore, the results indicated that Q808 treatment inhibited neuronal cell death, and significantly decreased the expression levels of apoptotic factors, including Apaf-1, caspase-9, and caspase-3. Q808 prevented neuronal damage in the hippocampus of TLE rats by targeting the intrinsic apoptotic pathway of neurons.

Microplastics induced inflammation and apoptosis via ferroptosis and the NF-κB pathway in carp

Microplastics (MPs), a new class of pollutant that threatens aquatic biodiversity, are becoming increasingly prevalent around the world. Fish growth may be severely inhibited by microplastics, resulting in severe mortality. Exposure to microplastics increases the likelihood of intestinal injuries, but the underlying mechanisms remain equivocal. The objective of this study was to investigate the potential toxic mechanisms underlying microplastic-induced intestinal injury in fish and to assist researchers in identifying novel therapeutic targets. In this study, a model of carp exposed to microplastics was established successfully. Histological observation showed that exposure to polyethylene microplastics caused damage to the intestinal mucosal surface and a significant increase in goblet cells, which aggregated on the surface of the mucosa. The mucosal layer was observed to fall off. Lymphocytes in the intestinal wall proliferated and aggregated. TUNEL staining showed that apoptosis occurred in the group exposed to microplastics. The qPCR results showed that the expression of Ferroptosis apoptotic factors COX-2 and ACSL4 was upregulated, while the expression of TFRC, FIH1, SLC7A11, and GPX4 was downregulated. The NF-κB pathway (p-p65, IκBα), inflammatory cytokines (TNF-α, IL-8, IL-6) and apoptosis genes (Bax, Caspase3) were upregulated. Semi-quantitative detection of related proteins by Western blotting was consistent with the gene expression results. In addition, the ELISA assay showed that lipid peroxidation and inflammatory cytokines (TNF-α, IL-1β, IL-6) were increased in the microplastic exposed group. To conclude, lipid peroxidation induced by microplastics activates the NF-κB pathway and causes ferroptosis, ultimately resulting in intestinal damage and cellular apoptosis.

Momordicae Semen inhibits migration and induces apoptotic cell death by regulating c-Myc and CNOT2 in human pancreatic cancer cells

Pancreatic cancer(PC) is less common than other cancers; however, it has a poor prognosis. Therefore, studying novel target signaling and anticancer agents is necessary. Momordicae Semen (MS), the seed of Momordica sochinensis Spreng, mainly found in South-East Asia, including China and Bangladesh, is used to treat various diseases because of its anticancer, antioxidant, anti-inflammatory, and antibacterial properties. However, the effect of the MS extract on pancreatic cancer cells remains unknown. In this study investigated whether the MS extract exerted an anti-cancer effect by regulating c-Myc through CNOT2. Cytotoxicity and proliferation were investigated using MTT and colony formation assays. The levels of apoptotic, oncogenic, and migration-associated factors were confirmed using immunoblotting and immunofluorescence. Wound closure was analyzed using a wound healing assay. The chemical composition of the MS methanol extracts was analyzed using liquid chromatography–mass spectrometry. We confirmed that the MS extract regulated apoptotic factors and attenuated the stability of c-Myc and its sensitivity to fetal bovine serum. Furthermore, the MS extract increased apoptosis by regulating c-Myc and CNOT2 expression and enhanced the sensitivity of 5-FU in pancreatic cancer. This study showed that the MS extract is a promising new drug for PC.