Expression Of Apoptosis-related Genes Research Articles

Integrating bioinformatics and machine learning to uncover lncRNA LINC00269 as a key regulator in Parkinson's disease via pyroptosis pathways

BackgroundPyroptosis, a specific type of programmed cell death, which has become a significant factor to Parkinson's disease (PD). Concurrently, long non-coding RNAs (lncRNAs) have garnered attention for their regulatory roles in neurodegenerative disorders. This study was designed to ascertain the key lncRNAs in pyroptosis pathways of PD and elucidate their regulatory mechanisms.MethodsEmploying a combination of bioinformatics and machine learning, we analyzed PD data sets GSE133347 and GSE110716. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) recognized different lncRNAs. Through various algorithms such as Least Absolute Shrinkage and Selection Operator (LASSO), Random Forest (RF), and Weighted Gene Co-expression Network Analysis (WGCNA), we recognized LINC01606 and LINC00269, which are key factors during the emergence and development of PD. Furthermore, experimental validation was conducted in PD mouse models to confirm these bioinformatics findings.ResultsThe analysis showed that there were a large number of apoptosis-related gene expression changes in Parkinson's syndrome, for example, CASP1 and GSDME were up-regulated, and CASP9 and AIM2 were down-regulated. Among the lncRNAs, LINC01606 and LINC00269 were identified as potential modulators of pyroptosis. Notably, LINC00269 was observed to be significantly downregulated in the brain tissues of a PD mouse model, supporting its involvement in PD. The study also highlighted potential interactions of these lncRNAs with genes like ONECUT2, PRLR, CTNNA3, and LRP2.ConclusionsThis study identifies LINC00269 as a potential contributor to pyroptosis pathways in PD. While further investigation is required to fully elucidate its role, these findings provide new insights into PD pathogenesis and suggest potential avenues for future research on diagnostic and therapeutic targets. The study underscores the importance of integrating bioinformatics with experimental validation in neurodegenerative disease research.

Effect of Liquid Blood Concentrates on Cell Proliferation and Cell Cycle- and Apoptosis-Related Gene Expressions in Nonmelanoma Skin Cancer Cells: A Comparative In Vitro Study.

Nonmelanoma skin cancer (NMSC) presents a significant challenge to global healthcare due to its rising incidence, prompting the search for innovative treatments to overcome the limitations of current therapies. Our study aims to explore the potential effects of the liquid blood concentrate platelet-rich fibrin (PRF) on basal cell carcinoma cells (BCCs) and squamous cell carcinoma cells (SCCs) in order to obtain results that may lead to new possible adjunctive therapies for managing localized skin cancers, particularly NMSC. Basal cell carcinoma (BCC) cells and squamous cell carcinoma (SCC) cells were indirectly treated with PRF generated via different relative centrifugation forces, namely high and low RCF PRF, for 7 days. PRF-treated cells were comparatively analyzed for cell viability, proliferation and cell cycle- and apoptosis-related gene expression. Analysis of MTS assay results revealed a significant decrease in cell viability in both BCC and SCC cells following PRF treatment for 7 days. Ki-67 staining showed a decreased percentage of Ki-67-positive cells in both BCC and SCC cells after 2 days of treatment compared to the control group. The downregulation of CCND1 gene expression in both cell types at 2 days along with the upregulation of p21 and p53 gene expression in SCC cells demonstrated the effect of PRF in inhibiting cell proliferation and inducing cell cycle arrest, especially during the initial phases of treatment. Increased expression of caspase-8 and caspase-9 was observed, indicating the activation of both extrinsic and intrinsic apoptotic pathways by PRF treatment. Although the exact immunomodulatory properties of PRF require further investigation, the results of our basic in vitro studies are promising and might provide a basis for future investigations of PRF as an adjunctive therapy for managing localized skin cancers, particularly NMSC.

Elaiophylin targets EIF4B to suppress the growth of esophageal squamous cell carcinoma via the PI3K/AKT signaling pathway.

Elaiophylin is known to exert antitumor effects through certain signaling pathways; however, no reports regarding its effects on esophageal cancer are available. This study explored the effects of elaiophylin in esophageal squamous cell carcinoma (ESCC) cells. Transwell and immunofluorescence assays confirmed that elaiophylin inhibited the migration and proliferation of ESCC cells, and western blotting assays showed that it affected apoptosis-related gene expression in ESCC cells. Based on RNA-seq analyses, Single-cell RNA-seq, a human cancer pathway phosphorylation antibody array, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomics analyses, we found that elaiophylin was related to low expression of EIF4B and activation of the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. In both in vitro and in vivo experiments, ESCC cells treated with elaiophylin showed low EIF4B expression, which inhibited their proliferation and promoted apoptosis by activating the PI3K/AKT signaling pathway; EIF4B overexpression could reverse these effects of elaiophylin on ESCC cells. Therefore, our results indicate that elaiophylin targets EIF4B to inhibit ESCC cell proliferation via the PI3K/AKT signaling pathway. Targeting elaiophylin or the EIF4B/PI3K/AKT signaling pathway may produce new methods for ESCC treatment.

Bushen Jianpi Tiaoxue Decoction (BJTD) ameliorates oxidative stress and apoptosis induced by uterus ageing through activation of the SIRT1/NRF2 pathway

BackgroundUterus ageing is a crucial factor contributing to decreased fertility in older women and is also implicated in menstrual disorders, endometritis, and adenomyosis. Bushen Jianpi Tiaoxue Decoction (BJTD) is a traditional Chinese medicine formulation used to ameliorate endocrine disorders in the female reproductive system and finds extensive application in ageing-related endometrial diseases. However, the mechanisms underlying its improvement of uterus ageing have not been thoroughly investigated. PurposeTo explore the potential components and mechanisms of BJTD in ameliorating uterus ageing through network pharmacology, in vivo, and in vitro experiments. MethodsMorphological changes were observed using hematoxylin and eosin staining, collagen deposition was assessed using Masson staining, and apoptotic-related molecules were detected using Western blot. After determining the modeling doses, BJTD intervention was administered at two doses, and the expression of oxidative stress and apoptosis-related genes and proteins was measured. The levels of cellular apoptosis were evaluated using the TUNEL assay kit and Annexin V/FITC-PI assay kit. The main components of BJTD were determined by UPLC-MS, and the potential targets and mechanisms of BJTD action were explored using network pharmacology and molecular docking. BJTD-Containing Serum (BJTD-S) was extracted and applied in vitro experiments using human endometrial stroma cells (hESC) to preliminarily identify the pathways affected. ResultsWe demonstrated that modeling with 600 mg/kg/day D-Gal for 5 weeks significantly increased collagen deposition in uterine tissues, particularly in the glands and stroma. Additionally, it significantly elevated the levels of TNF-α and IL-1β and increased the expression of p53 and BAX while decreasing BCL-2 expression. BJTD significantly reduced the increased levels of TNF-α and IL-1β induced by D-Gal, and modulated oxidative stress markers such as SOD, MDA, GSH-Px, and T-AOC. BJTD also inhibited the cascade activation of apoptosis induced by D-Gal, suppressing the expression of cleaved-Caspase 8, cleaved-Caspase 3, and BAX. SIRT1 is a potential target of BJTD action. In vitro experiments showed that BJTD-S significantly improved D-Gal-induced apoptosis in hESC cells, and the expression levels of SIRT1, NRF2, and HO-1 were significantly decreased in D-Gal-induced hESC, and BJTD-S significantly increased their expression. ConclusionBJTD can ameliorate oxidative stress and cell apoptosis levels in D-Gal-induced uterine aging, and its active ingredients can activate the SIRT1/NRF2 pathway to exert its effects. Importantly, our study provides novel insights into the molecular mechanisms by which traditional Chinese medicine influence uterus ageing. By specifically targeting the SIRT1/NRF2 pathway, BJTD presents a unique therapeutic approach that has not been extensively explored in previous studies, marking a significant advancement in the treatment of uterus ageing.

Lactiplantibacillus plantarum Ameliorated Morphological Damage and Barrier Dysfunction and Reduced Apoptosis and Ferroptosis in the Jejunum of Oxidatively Stressed Piglets.

Oxidative stress induces apoptosis and ferroptosis, leading to intestinal injury of piglets. Lactiplantibacillus plantarum P8 (P8) has antioxidant capacity, but its roles in intestinal apoptosis and ferroptosis remain unclear. Here, 24 weaned piglets were assigned to three treatments: control (Con), diquat injection (DQ), and P8 supplementation + DQ injection (DQ + P8). The results showed that the increased jejunal oxidative stress, jejunal morphology impairment, and barrier dysfunction in the DQ-treated piglets were decreased by P8 supplementation. TUNEL and apoptosis-related gene expressions showed increased jejunal apoptosis of DQ-treated piglets; however, reduced apoptosis was observed in the DQ + P8 group. In addition, the mitochondrial morphology and ferroptosis-related gene expressions indicated elevated jejunal ferroptosis in the DQ-treated piglets, and the DQ + P8 treatment attenuated the ferroptosis. Transcriptome identified various differentially expressed genes (DEGs) between different treatments. KEGG analysis indicated that the DEGs were enriched in the PI3K-AKT, NF-κB, and apoptosis pathways. The expressions of key DEGs and key proteins in the PI3K-AKT and NF-κB pathways were further verified. In summary, our results indicate that P8 supplementation ameliorated jejunal oxidative stress, morphological damage, barrier dysfunction, apoptosis, and ferroptosis in the DQ-treated piglets. Moreover, the beneficial effect of P8 may be related to the regulation of PI3K/AKT and NF-κB pathways.

Comparison of the Mitophagy and Apoptosis Related Gene Expressions in Waste Embryo Culture Medium of Female Infertility Types.

Mitochondria is an important organelle for the oocyte-to-embryo transition in the early embryonic development period. The oocyte uses mitochondria functionally and its mitochondrial DNA (mtDNA) content as the main energy source in the embryo development at the preimplantation stage. The aim of this study is to compare mitophagic, apoptotic and humanin gene expressions from the culture medium fluid in which embryos are developed and monitored among normoresponder (NOR), polycystic ovary syndrome (PCOS), young and older patients with poor ovarian reserve (POR). The study groups consisted of infertile patients who applied to the Bahçeci Umut IVF Center as NOR (Control), PCOS, POR-Advanced (POR-A) and POR-Young (POR-Y). After the isolation of total RNA from the collected samples, MFN1, MFN2, PINK1, PARKIN, SMN1, SMN2, p53 and Humanin gene expressions were determined by Real Time-PCR. The average age of only the POR-A was determined to be higher than the NOR (p < 0.001). The MFN1, SMN2 (p < 0.05), Humanin and p53 gene expressions (p < 0.001) increased, while PINK1 gene expression decreased (p < 0.05), in the POR-Y compared to the NOR. A decrease in MFN2, PARKIN (p < 0.05) and PINK1 gene expressions was determined in the PCOS compared to the NOR (p < 0.001). Furthermore, a decrease was observed in MFN2, PINK1 (p < 0.001) and Humanin gene expressions compared to the NOR (p < 0.05). The current data are the first in the literature determining the apoptotic and mitophagic status of the oocyte. The current results prove that waste embryo culture fluid may provide a non-invasive profile for important cellular parameters such as mitochondrial dysfunction in female infertility. The evaluation of significant cellular parameters can be performed much earlier without any intervention into the embryo.

Soy\'s secret weapon: Genistein\'s fight against triple-negative breast cancer

Triple-negative breast cancer (TNBC) is an aggressive form of breast cancer with limited treatment options. Genistein a natural isoflavone found in soybeans and legumes is a plant-based compound with anticancer properties that has been shown to be promising for TNBC treatment in preclinical studies. This review examines genistein’s potential as a therapeutic agent against TNBC. In vitro studies reveal its capacity to inhibit cell growth, induce apoptosis, and suppress TNBC metastasis. In vivo, genistein suppresses tumor growth and extends survival in TNBC mice. It targets key oncogenic pathways, including NF-κB, Akt, and PI3K. It has also been shown to regulate apoptosis-related gene expression, promote apoptosis, and activate the DNA damage response. Furthermore, Genistein demonstrated the ability to reactivate estrogen receptor expression in ERα-negative MDA-MB-231 xenograft mice, particularly when combined with the histone deacetylase inhibitor trichostatin A. This reactivation restored ER-dependent cellular sensitivity to tamoxifen and 17-estradiol. Combination therapy with genistein and other drugs or phytochemicals has shown to be particularly effective in preclinical studies. These findings suggest that Genistein holds promise as a potential therapeutic agent for TNBC by targeting key signaling pathways involved in cell proliferation, survival, and apoptosis, as well as modulating ER expression and enhancing sensitivity to existing therapies.

Ferulic Acid Protects Human Lens Epithelial Cells Against UVA-Induced Oxidative Damage by Downregulating the DNA Demethylation of the Keap1 Promoter.

Ultraviolet (UV) radiation-triggered production of reactive oxygen species (ROS) is a primary contributor to apoptosis in human lens epithelial cells (HLECs), which can ultimately result in cataract formation. The nuclear factor erythroid-2-related factor 2 (Nrf2)-Kelch ECH associating protein 1 (Keap1) pathway, a fundamental oxidative stress regulation mechanism, plays a crucial role in the development of cataracts. Ferulic acid (FA), recognized for its potent antioxidant properties can activate the Nrf2 signaling pathway to mitigate oxidative damage and cell apoptosis. In this study, we have demonstrated the protective effects of FA in reducing UVA-induced oxidative damage and apoptosis in HLECs through the modulation of the Keap1/Nrf2 pathway, as evidenced by both cellular and animal experiments. HLECs and Lens were exposed to 10 J/cm2 UVA radiation with or without prior treatment with FA. We found that UVA radiation increased oxidative damage and cell apoptosis in HLECs, ultimately leading to opacification of rat lenses, while FA was able to attenuate both oxidative damage and cell apoptosis in HLECs and reduce the degree of lens opacification. FA upregulated the expression of antioxidant response factors of the Keap1/Nrf2 pathway and downregulated the expression of apoptosis-related genes in HLECs, as demonstrated by Western blot and RT-qPCR analyses. We also found that UVA radiation increased the degree of demethylation of the Keap1 promoter in HLECs, whereas FA reduced the level of Keap1 promoter demethylation as determined by DNA sequencing. Additionally, UVA upregulated the expression of DNA active demethylase of the Keap1 promoter in HLECs, Dnmt1, Dnmt3a, and Dnmt3b, as shown by immunofluorescence, Western blot, and RT-qPCR, however, FA attenuated the activity of the passive demethylase TET1 in addition to the active demethylases. These results demonstrated that UVA radiation can cause oxidative damage, cell apoptosis, and rat lens opacification by increasing the demethylation of the Keap1 promoter in lens epithelial cells. Conversely, FA was shown to reduce oxidative damage, inhibit cell apoptosis, and decrease rat lens opacification by increasing the methylation of the Keap1 promoter. These findings suggest that FA could be therapeutically beneficial in preventing and mitigating cataracts induced by UVA radiation.

Study on the role of RPL23 gene in active immunity of termite Reticulitermes chinensis against Metarhizium anisopliae

Ribosomal proteins are considered to be involved in the immunity of different animals against pathogens. The protein level of RPL23 increased after fungal infection in termites, but how it influence active immunity in termites is unknown. The role of RPL23 gene was studied to evaluate its impact on active immunity of termite Reticulitermes chinensis against entomopathogenic fungus (EPF) Metarhizium anisopliae. The RPL23 gene fragment (414 bp) was cloned and phylogenetic analysis revealed that it’s very close to termite Coptotermes formosanus. Expression of RPL23 gene was significantly higher in abdomen as compared to thorax and head. Silencing RPL23 gene had no significant impact on the frequency and time of allogrooming towards fungus exposed termites from nestmates, which showed that nestmates acquired spores from infected termites through allogrooming. Expression of immune genes (GNBP1, GNBP2 and phenoloxidase) and apoptosis related genes (TNF-α, caspase 1, caspase 3 and caspase 8) decreased significantly in nestmates of fungus-treated termites after silencing of RPL23 gene as compared to control. Antifungal activity and survival of RPL23 silenced nestmates of fungus-treated termites also decreased. To sum up, this study found that silencing of RPL23 gene broke the active immunity against M. anisopliae infection, reduced the antifungal activity of termites, weakened cell apoptosis, and led to increased mortality of termites, which may help to find a potential alternative for chemical insecticides to control termites.

The Effects of Crude Alkaloid Extracts of Convolutes Arvensis on Tumour Cell Lines and Their Potential For Toxicity

General Background: Cancer remains a leading cause of mortality worldwide, and the search for effective treatments continues to intensify. Specific Background: Convolvulus arvensis has been traditionally used for medicinal purposes, but its potential as an anticancer agent is underexplored. Knowledge Gap: The cytotoxic and apoptotic mechanisms of crude alkaloids extracted from C. arvensis against specific cancer cell lines have not been fully characterized. Aims: This study investigates the antioxidant activity and cytotoxic effects of C. arvensis crude alkaloids on mouse liver cancer (HC) and human breast cancer (AMJ13) cell lines, focusing on apoptosis-related gene expression. Results: The antioxidant activity of C. arvensis was comparable to that of ascorbic acid, with inhibition rates of 92.01% at 500 µg/ml. Crude alkaloids demonstrated dose-dependent cytotoxicity, with a maximum inhibition rate of 82.65% in AMJ13 cells and 79.49% in HC cells at 500 µg/ml. Gene expression analysis revealed upregulation of caspase-9, indicating apoptosis via the intrinsic mitochondrial pathway. Novelty: This study is among the first to provide molecular evidence of C. arvensis-induced apoptosis through the intrinsic pathway, offering a novel insight into its anticancer potential. Implications: These findings suggest that C. arvensis alkaloids could be developed as a therapeutic option for cancer treatment, with future studies needed to isolate specific compounds and assess their in vivo efficacy. Highlights: C. arvensis alkaloids show antioxidant activity similar to ascorbic acid. Alkaloids inhibit liver and breast cancer cell growth dose-dependently. Apoptosis triggered via caspase-9 through the mitochondrial pathway. Keywords: Convolvulus arvensis, alkaloids, cancer, apoptosis, antioxidant

CREBRF regulates apoptosis and estradiol via ISG15/ISGylation in pig granulosa cells

Granulosa cells play a crucial role in the reproductive processes of female animals, as their proliferation, apoptosis, and hormonal secretion are vital for follicular development and ovulation. Although the role and mechanisms of CREBRF in the reproductive system have been partly reported, its functions in ovarian granulosa cells have not been fully explored. In this study, the results indicated that the expression of CREBRF in the ovaries at 30 days after birth was significantly higher than that during puberty and sexual maturity. Studies on the function of CREBRF found that CREBRF could enhance the synthesis of estradiol and had no effect on progesterone synthesis in pig granulosa cells. At the same time, CREBRF could suppress apoptosis through the Bax/caspase3/caspase9 pathway and modulation of ISG15/ISGylation in pig granulosa cells. During this process, the expression of many genes changed in granulosa cells. Several genes (CMPK2, MX1, MX2, ZBP1, PML, CHAC1, and BAX) which were promoted apoptosis, were upregulated after CREBRF knockdown with siRNA. ISG15-protein conjugation genes (HERC5, UBA7, UBE2L6, ISG15) were also were upregulated. On the contrary, the expression of anti-apoptotic (RFK, SNAP23) genes decreased. In conclusion, CREBRF could enhance the synthesis of estradiol and acted as anti-apoptosis role in pig granulosa cells. This discovery can provide novel insights for further elucidating the molecular mechanisms of granulosa cells in the ovary and potentially identifies CREBRF as a molecular target for improving fertility.

Mitoquinone improves porcine embryo development through modulating oxidative stress and mitochondrial function

Oxidative stress caused by excess reactive oxygen species (ROS) is one of the main causes of low efficiency in in vitro production of embryos. These ROS can cause mitochondrial dysfunction and apoptosis, resulting in poor embryo development. Therefore, to prevent mitochondrial damage and apoptosis caused by ROS, we investigated the effects of mitoquinone (MitoQ), a mitochondrial-targeted antioxidant, on the in vitro culture (IVC) of porcine embryos. Various concentrations of MitoQ (0, 0.01, 0.1, or 1 nM) were supplemented during the entire period of IVC. The results showed that supplementation with 0.1 nM MitoQ significantly increased the blastocyst formation rate, with a higher total cell number including trophectoderm cell number and higher transcript expression of lineage-specific transcription factors in blastocysts. In addition, the 0.1 nM MitoQ-treated group showed a significantly lower percentage and number of apoptotic cells in blastocysts with positively regulated transcript expression of apoptosis-related genes. Therefore, 0.1 nM MitoQ was suggested as optimal concentration for porcine IVC and used for further investigations. MitoQ treatment significantly reduced intracellular ROS levels and increased glutathione levels in Day 2 embryos, with upregulated the transcript expression of antioxidant enzymes-related genes. Furthermore, the MitoQ group exhibited a significantly higher mitochondrial quantity, mitochondrial membrane potential, and ATP content in Day 2 embryos, with increased transcript expression of mitochondrial biogenesis-related genes. Taken together, these findings reveal that MitoQ supplementation can enhance the developmental competence of porcine embryos by decreasing oxidative stress and improving mitochondrial function.

Apoptosis-stimulating protein of p53 (ASPP) participates in the regulation of apoptosis in Litopenaeus vannamei under ammonia-N and nitrite-N stress

Apoptosis-stimulating protein of p53 (ASPP) is a key regulatory factor closely related to p53 in apoptosis pathway. To further investigate the molecular mechanisms of ASPP in Litopenaeus vannamei, the expression of LvASPP mRNA under ammonia-N and nitrite-N stress was explored, and the effects of knocking out LvASPP on mortality, histological damage, and the apoptosis pathway in L. vannamei under ammonia-N and nitrite-N stress were investigated. Healthy L. vannamei (7.78 ± 0.70 g) were used in this study. After shrimp were stressed with an ammonia-N concentration of 30.00 mg/L for 48 h, qRT-PCR was used to detect a significant increase in LvASPP mRNA expression in the hepatopancreas, gills, and muscle. Following 48 h of nitrite-N stress at a concentration of 60.00 mg/L, LvASPP mRNA expression was significantly increased in the hepatopancreas. The survival rate notably increased under 80 h of ammonia-N stress (25.00 mg/L) after LvASPP RNA interference (30 % more than the control group), and the number of shrimp deaths decreased after 48 h of nitrite-N stress. Moreover, under the stress of ammonia-N and nitrite-N respectively, LvASPP silencing reduced the expression of p53, and led to a decrease in the expression of apoptosis-related genes (Bax, Apaf-1, Caspase 9, MDM2). Caspase 3 activity, TUNEL-positive cells and the apoptotic index in the hepatopancreas markedly reduced under ammonia-N and nitrite-N stress. The potential pathway suggests that inhibiting LvASPP reduces the mRNA expression of p53, which leads to a decrease in Caspase 3 activity, inhibiting apoptosis in the hepatopancreatic cells of L. vannamei under ammonia-N and nitrite-N stress. These data indicate that the knockdown of LvASPP positively impacted the tolerance of L. vannamei to ammonia-N and nitrite-N stress by regulating the apoptosis pathway. This suggests that employing gene-targeted dsRNA could be an effective strategy for alleviating the environmental pressures faced by shrimp in aquaculture management.

Acupuncture improves spatial learning and memory impairment caused by herpes simplex virus type-1 in rats through the p38 MAPK/CREB pathway

BackgroundAcupuncture can improve herpes simplex encephalitis (HSE), but the underlying mechanism is not clear. Therefore, we evaluated the cognitive function and apoptosis in hippocampus caused by herpes simplex virus type-1 (HSV-1) in rats after acupuncture and described the molecular mechanism.MethodsSprague–Dawley rats were induced into HSE models by HSV-1 infection. After 3 days, they received acupuncture at the acupoints of Xuanzhong (GB39), Baihui (GV20), Shenmen (HT7), Shenting (GV24), and Sanyinjiao (SP6), and/or intraperitoneal injection of the p38 MAPK inhibitor SB203580. Morris water maze test was performed on rats. The hippocampus of rats was obtained, and the expression of apoptosis-related genes in the tissues was detected by qRT-PCR. In addition, apoptosis-related proteins and proteins related to the p38 MAPK/CREB pathway in the tissues was detected by western blot.ResultsAfter HSV-1 induction, the rat's escape latency was increased, the time spent on the platform in the target quadrant and the number of platform crossings significantly decreased. In addition, there was an increase in apoptosis in the hippocampus, accompanied by elevated levels of p–p38 and decreased levels of p-CREB. However, these effects could be improved by acupuncture treatment. Interestingly, SB203580 plays a similar role to acupuncture, and acupuncture could further enhance the impacts of SB203580 on cognitive function and apoptosis in hippocampus in HSE rats.ConclusionAcupuncture improves spatial learning and memory impairment caused by HSV-1 in rats. The functional mechanism of acupuncture may be through the p38 MAPK/CREB pathway.

DNA damage-inducible transcript 3 positively regulates RIPK1-mediated necroptosis.

DNA damage-inducible transcript 3 (DDIT3) is a well-known transcription factor that regulates the expression of apoptosis-related genes for promoting apoptosis during endoplasmic reticulum stress. Here, we report an unrecognized role of DDIT3 in facilitating necroptosis. DDIT3 directly binds and competitively prevents the p38 MAPK-MK2 interaction and thereby blocking MK2 activation while stimulating p38 MAPK activation. This blockage of MK2 activation initially prevents RIPK1 phosphorylation at Ser320 (inactivation), subsequently relieving its suppression of RIPK1 activation. Consequently, p38 MAPK facilitates RIPK1 phosphorylation at Ser166 (activation) through DDIT3 phosphorylation-related mechanisms, leading to necroptosis. Mechanistically, a 10-amino acid segment (Glu19-Val28) within DDIT3's N-terminus is identified to account for its pro-necroptotic function. In vivo studies demonstrate that forced expression of DDIT3 induces necroptosis, whereas deletion of DDIT3 alleviates necroptosis in mouse hearts under stress. These findings shed light on a novel regulatory mechanism by which DDIT3 promotes RIPK1 activation and subsequent necroptosis.

Damage to Mitochondria During the Cryopreservation, Causing ROS Leakage, Leading to Oxidative Stress and Decreased Quality of Ram Sperm.

Semen cryopreservation can achieve long-term preservation of sperm. Ice crystal damage, as well as oxidative stress, result in mitochondrial dysfunction and a reduction in sperm motility after thawing. However, limited information exists regarding the impact of reactive oxygen species (ROS) and mitochondria on the cryopreservation of ram sperm. The primary objective of this study was to investigate the relationship between ROS and mitochondria concerning sperm quality during the cryopreservation of ram sperm. This investigation assessed sperm motility, kinematic characteristics, membrane integrity, acrosome integrity, mitochondrial membrane potential (MMP), adenosine triphosphate (ATP) levels, expression of mitochondrial respiratory genes (NDUFV2, SDHA, CYC1, and COXIV), ROS levels, malondialdehyde (MDA) content, phosphatidylserine externalisation rate, sperm ultrastructure, mtDNA copy number, expression of apoptosis-related genes (Bax, Caspase-3, and Caspase-8), Cytochrome C, and Caspase-3 content. The results showed the cryopreservation significantly (p < 0.05) decreased motility, kinetic parameters, membrane integrity, acrosome integrity, MMP, ATP, mRNA expression levels of mitochondrial respiratory-related genes, and significantly (p < 0.05) increased ROS levels, MDA content, phosphatidylserine externalisation rate, damage of sperm ultrastructure, mtDNA copy number, mRNA expression levels of apoptosis-related genes, Cytochrome C and Caspase-3 content compared to the fresh semen group. In conclusion, the cryopreservation causes damage to mitochondria, leading to increased ROS and subsequent oxidative stress. This process also initiates mitochondrial dysfunction and interferes with the electron transport chain, ultimately resulting in decreased MMP and ATP production. Furthermore, the liberation of Cytochrome C prompted the increase in Caspase-3 expression and subsequent sperm apoptosis occurred, ultimately leading to a deterioration in sperm quality after thawing.

Protective Effect of Endogenous ω-3 Polyunsaturated Fatty Acid Against Cisplatin Induced Myelosuppression

To investigate the protective effect of endogenous ω-3 polyunsaturated fatty acid (PUFA) against cisplatin-induced myelosuppression and the mechanism of reducing apoptosis in bone marrow nucleated cells using mfat-1 transgenic mice. The experimental animals were divided into 4 groups: wild-type mice normal control group, mfat-1 transgenic mice normal control group, wild-type mice model group and mfat-1 transgenic mice model group. The mice in the model group were injected intraperitoneally with 7.5 mg/kg cisplatin on day 0 and day 7 to construct a myelosuppression model, while the mice in the normal control group were injected intraperitoneally with an equal amount of saline, and their status was observed and their body weight was measured daily. Peripheral blood was taken after 14 day for routine blood analysis, and the content and proportion of PUFA in peripheral blood were detected using gas chromatography. Bone marrow nucleated cells in the femur of mice were counted. The histopathological changes in bone marrow were observed by histopathological staining. The apoptosis of nucleated cells and the expression level changes of apoptosis-related genes in the bone marrow of mice were detected by flow cytometry and fluorescence quantitative PCR. Compared with wild-type mice, mfat-1 transgenic mice showed significantly increased levels of ω-3 PUFA in peripheral blood and greater tolerance to cisplatin. Peripheral blood analysis showed that endogenous ω-3 PUFA promoted the recovery of leukocytes, erythrocytes, platelets and haemoglobin in peripheral blood of myelosuppressed mice. The results of HE staining showed that endogenous ω-3 PUFA significantly improved the structural damage of bone marrow tissue induced by cisplatin. Flow cytometry and PCR showed that, compared with wild-type mice model group, the apoptosis rate of bone marrow nucleated cells in mfat-1 transgenic mice was significantly reduced ( P < 0.001), and the expression of anti-apoptotic genes Bcl-2 mRNA was significantly increased ( P < 0.01), while the expressions of pro-apoptotic genes Bax and Bak mRNA were significantly reduced ( P < 0.001, P < 0.05). Endogenous ω-3 PUFA can reduce cisplatin-induced apoptosis in bone marrow nucleated cells, increase the number of peripheral blood cells and exert a protective effect against cisplatin-induced myelosuppression by regulating the expression of apoptosis-related genes.

LncRNA Pvt1 aggravates cardiomyocyte apoptosis via the microRNA-216/Ccnd3 axis

ObjectiveOur study aims to evaluate the role of long non-coding RNA variant translocation gene Pvt1 in cardiomyocyte apoptosis, as well as the potential targets and mechanisms involved in Pvt1-miRNA-mRNA axis. Methods1.Pvt1 knockdown in cells by transfection with small interfering RNA (si-Pvt1), HL-1 cells were randomly divided into control group, hypoxia group, hypoxia + negative control group and hypoxia + si-Pvt1 group. Apoptosis-related genes expression was detected by Western blot assay, RT-qPCR and Flow cytometry assay. 2.Pvt1 knockdown model (sh-Pvt1) was established by injecting adeno-associated virus (AAV) vector shRNA-Pvt1 into the caudal vein 7 days before myocardial infarction, and echocardiography was used to measure cardiac function 7 days after myocardial infarction induced by ligation of the left anterior descending branch. HE staining was used to evaluate the pathological injury of mouse heart tissue, and the apoptotic protein expression was detected by Western blot. 3.lncRNA-related microRNAs were predicted by bioinformatics tools and further verified by dual luciferase experiment. Western blot analysis was used to identify the expression of apoptotic genes following the simultaneous transfection of si-Pvt1 and miR-216 mimics. Genes differentially expressed in hypoxia + si-NC and hypoxia + si-Pvt1 groups were identified by RNA sequencing. These genes were then compared with the target genes of miR-216 predicted by bioinformatics tools. The gene of interest Ccnd3 was excluded from the analysis. Western blot analysis was used to assess the expression of Apoptosis-related proteins in HL-1 cells co-transfected with miR-216 mimics and overexpressed Ccnd3. Results1. Pvt1 was highly expressed in HL-1 induced by hypoxia, and Pvt1 knockdown can reduce cell apoptosis in hypoxia cells. 2. MI causes myocardial injury in mice, and inhibition of Pvt 11 can improve the cardiac function of mice with myocardial infarction, prevent some inflammatory cell infiltration, and reduce myocardial cell apoptosis. 3. Pvt1 acts as a sponge for miR-216 and promotes the expression of Ccnd3. ConclusionPvt1 may promote myocardial infarct-induced apoptosis through the miR-216/Ccnd3 axis.

Enhanced understanding of cinnamaldehyde's therapeutic potential in osteoarthritis through bioinformatics and mechanistic validation of its anti-apoptotic effect.

Osteoarthritis (OA) is a globally prevalent joint disorder affecting approximately 240 million individuals worldwide. Cinnamaldehyde, known for its broad anti-inflammatory and anti-aging effects across various cell types, has not been investigated for its potential impact on apoptosis in OA chondrocytes. To explore the effectiveness of cinnamaldehyde in mitigating knee osteoarthritis by reducing chondrocyte apoptosis, bioinformatics analysis was first conducted to identify apoptosis-associated differentially expressed genes (APDEGs). Gene expression datasets GSE55235 and GSE114007 were analyzed using weighted gene co-expression network analysis (WGCNA). Gene modules of interest were cross-referenced with APDEGs to identify those specific to OA. LASSO regression analysis was employed to build a risk model, and this model, along with datasets GSE114007, GSE55457, and GSE12021, was validated using ROC analysis. Cellular experiments and blood analyses from OA patients were performed to evaluate the effects of cinnamaldehyde on apoptosis-related gene expression. Cinnamaldehyde administration was found to rectify the abnormal expression of key apoptosis-related genes in OA patients. Specifically, cinnamaldehyde may affect knee osteoarthritis by regulating apoptosis-related genes such as ZFAND5, BCL6, ELL2, FOSL2, MARCKS, and SGCD. Additionally, three novel apoptotic targets in OA chondrocytes-ZFAND5, ELL2, and SGCD-were identified. These findings provide significant theoretical support for the clinical use of cinnamaldehyde in OA treatment. The discovery of novel apoptotic targets presents new therapeutic possibilities for future OA interventions.

Dietary Addition of Selenium Attenuates Cadmium-Induced Liver Injury in Grass Carp (Ctenopharyngodon idellus) by Reducing Oxidative Stress and Apoptosis

In order to investigate the effects of selenium (Se) against cadmium (Cd) toxicity, 180 healthy grass carp were separated into three groups and fed diets containing 0.147 (control group), 0.562, and 1.044 mg/kg of selenium Yeast throughout 60 days. In grass carp livers, malondialdehyde (MDA) content, antioxidant enzyme activities, and apoptosis-related gene expression were examined. As a result of acute exposure to cadmium, MDA content decreased significantly. With time, catalase (CAT), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px) activities changed. The relative transcript levels of heavy metal scavenging genes abcc2 and mt2 were significantly reduced. The relative levels of expression of jnk, bax, caspase-3, caspase-8, and caspase-9 in apoptosis-associated factors were significantly elevated after cadmium exposure. Selenium-supplementation downregulated the expression of apoptosis-related factors. As compared to the control group, liver cells supplemented with selenium had a significantly lower apoptotic index. Comprehensive analysis showed that dietary selenium supplementation significantly attenuated cadmium-induced peroxidative damage and apoptosis in liver by increasing GSH-Px activity, and that cadmium toxicity could be alleviated by the addition of yeast selenium.