Stress Protein Levels Research Articles

Sinapic acid alleviates glutamate-induced excitotoxicity by inhibiting neuroinflammation and endoplasmic reticulum stress pathway in C6 glioma cells

Sinapic acid (SA) is a polyphenol compound derived from hydroxycinnamic acid found in various foods such as cereals and vegetables and has antioxidant, anti-inflammatory and neuroprotective properties. However, its effects on glutamate-induced excitotoxicity, which is important in neurodegenerative diseases, have not been fully elucidated. This study aimed to investigate the effect of SA on glutamate excitotoxicity and the possible role of proinflammatory cytokines and the endoplasmic reticulum (ER) stress pathway. In the study, C6 rat glioma cell line was used and the cells were divided into 4 groups: control, glutamate, SA and glutamate+SA. Cells were treated with 10 mM glutamate for 24 h to induce excitotoxicity. Additionally, SA was applied to cells at concentrations of 12.5 to 100 μM to examine its effects on glutamate excitotoxicity. XTT test was used for cell viability, and apoptotic cells were determined by immunofluorescence and flow cytometry methods. Proinflammatory cytokines (tumor necrosis factor-alpha, TNF-α and interleukin-beta, IL-1β), ER stress markers (glucose regulatory protein 78, GRP78; C/EBP homologous protein, CHOP and activating transcription factor-4, ATF-4) and caspase-3 was used to measure ELISA method. Findings indicated that SA (50 μM) significantly increased cell viability against glutamate-induced excitotoxicity (p < 0.05). Also, SA caused a significant decrease in TNF-α, IL-1β, GRP78, CHOP, ATF-4 and caspase-3 levels in glutamate-treated cells (p < 0.05). Flow cytometry and immunofluorescence staining results showed that SA reduced apoptosis in C6 glioma cells. In conclusion, our findings suggested that SA attenuated glutamate-induced excitotoxicity by preventing apoptosis through reducing proinflammatory cytokines and ER stress protein levels.

Ficus deltoidea standardized methanolic extract improves kidney structure and function against streptozotocin-induced diabetic nephropathy in rats

BackgroundOxidative stress and peripheral insulin resistance are the key pathogenesis of diabetic nephropathy. We have previously demonstrated that Ficus deltoidea increases insulin secretion and improves tissue regeneration by reducing oxidative stress in diabetic rats. However, it remains uncertain whether F. deltoidea could delay the progression of nephropathy. Hypothesis/PurposeThis study aimed to examine the nephroprotective effects of F. deltoidea on streptozotocin-induced diabetic nephropathy in rats. MethodsHigh-Performance Liquid Chromatography (HPLC) was used to identify the flavonoid compounds in the leaf extract of F. deltoidea. The methanolic extract of F. deltoidea was administered orally at 1000 mg/kg body weight for eight weeks to diabetic rats. Serum creatinine, urea, uric acid, total bilirubin, and urinary creatinine were measured to estimate kidney function. Enzyme-linked immunosorbent assays (ELISA) assessed the levels of oxidative stress, antioxidants, and apoptosis-related proteins in the kidney tissue. The spectral markers related to nephroprotective activity were predicted using Fourier-transform infrared (FTIR) spectroscopy combined with chemometric analysis. Histomorphometric evaluations were performed on the kidney sections stained with hematoxylin and eosin (H&E). ResultsHPLC identified the presence of vitexin, isovitexin, quercetin, and kaempferol in F. deltoidea methanolic leaves extract. The biochemical and histological examinations consistently showed that F. deltoidea extract improved kidney structure and function by reducing oxidative stress and apoptosis. We noticed that biochemical and pathological changes corresponded to the infrared (IR) peaks at 1545 cm-1 and 1511 cm-1 (amide II), 1246 cm-1 (DNA/RNA phosphate backbones), and 1181 cm-1 (C–O vibrations). Multivariate analysis of IR spectra demonstrated that the diabetic rats treated with F. deltoidea extract had a similar clustering pattern to that of the normal animals. ConclusionThese findings indicate that the methanolic extract of F. deltoidea exhibits nephroprotective activity. FTIR spectroscopy may be useful for monitoring structural and biochemical alterations in the kidney during diabetes treatment.

The Neuroprotective Effects of Peripheral Nerve Microcurrent Stimulation Therapy in a Rat Model of Middle Cerebral Artery Occlusion.

This study investigated the neuroprotective effects of peripheral nerve microcurrent stimulation therapy in a rat model of middle cerebral artery occlusion (MCAO). Twenty 8-week-old male Sprague Dawley rats weighing 300-330 g were categorised into group A, serving as the healthy control; group B, including rats subjected to MCAO; group C, including rats receiving microcurrent therapy immediately after MCAO, which was continued for one week; and group D, including rats receiving microcurrent therapy one week before and one week after MCAO. A gross morphological analysis, behavioural motion analysis, histological examination, immunohistochemistry, and Western blotting were conducted. Microcurrent therapy significantly reduced ischaemic damage and pyramidal cells of the hippocampus CA1 region. Haematoxylin and eosin staining revealed infarction areas/viable pyramidal cell numbers of 0%/94.33, 28.53%/40.05, 17.32%/80.13, and 5.38%/91.34 in groups A, B, C, and D, respectively (p < 0.001). A behavioural analysis revealed that the total distances moved were 1945.24 cm, 767.85 cm, 1781.77 cm, and 2122.22 cm in groups A, B, C, and D, respectively (p < 0.05), and the mean speeds were 6.48 cm/s, 2.50 cm/s, 5.43 cm/s, and 6.82 cm/s, respectively (p < 0.05). Inflammatory markers (cluster of differentiation 68, interleukin-6, and tumour necrosis factor-α) significantly decreased in the treated groups (p < 0.001). Western blotting revealed reduced proinflammatory, oxidative stress, and apoptosis-related protein levels, along with increased angiogenic factors and mitogen-activated protein kinase (MAPK) pathway modulation in the treated groups. Peripheral nerve microcurrent stimulation therapy effectively mitigates ischaemic damage, promotes recovery, reduces inflammation, and modulates protein expression, emphasising its potential as a therapeutic strategy for ischaemic stroke.

Effect of propolis applied to goat kids at weaning period on heat shock protein genes

Weaning is a stressful period that affects the welfare of goat kids. During this period, goat kids lose weight due to reduced food intake, and they become susceptible to viral and bacterial infections. In recent years, many studies have been carried out on more natural and organic additives to prevent the loss of offspring in goat breeding and to promote their growth and development by limiting the use of antibiotics. Propolis has potent immunomodulatory, antioxidant, and anti-inflammatory properties. We aimed to investigate the effects of propolis on weaning stress in this study. The expression levels of the molecular chaperones which modulate the cellular stress response HSP27, HSP60, and HSP70 were examined. The Saanen kids were divided into propolis-treated (n=10) and control (non-propolis treated; n=10) groups. The propolis-treated group was given 0.4 cc of propolis once daily for 2 weeks. Expression levels were calculated by 2 -ΔΔCt using the Pfaffl method in the sample taken on the day after weaning and at the end of the 2-week propolis application. During weaning stress in kids, the expression levels of HSP27 and HSP60 were increased by 2-fold or more while HSP70 was upregulated by 3.57 fold. When 0.4 cc of propolis was administered to kids under weaning stress, there was a statistically significant down-regulation of 0.99-fold in the HSP27, 1.53-fold in the HSP60 level and 2.61-fold in HSP70 level. Our study showed that propolis application reduced stress protein levels during weaning stress.

Potential protective role of chlorogenic acid against cyclophosphamide-induced reproductive damage in male mice.

Cyclophosphamide (CP) is an anticancer drug; however, clinical utilization of CP is limited, resulting from its considerable toxicities. This research was performed to explore the protective effects of Chlorogenic acid (CGA) on reproductive damage induced by CP in mice. Blood samples were collected for analysis of hormone content subsequently; semen samples were evaluated for quality, and testis samples were used for histopathological evaluation and analysis of oxidative stress biomarkers, protein and gene expression levels of steroid regulatory factors, and steroid synthase. The results noted that CGA increased serum testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) activity; increased SOD, GPx, and GSH oxidative stress levels in testis tissue; and decreased MDA content in testis tissue. Testicular cells in the CGA treatment group gradually returned to normal morphology, and CYP11A1 and CYP17A1 levels increased after CGA treatment. The mRNA levels of CYP11A1, CYP17A1, StAR, 3β-HSD, and 17β-HSD were significantly raised in the CGA dose group. In the test dose range, CGA can improve sperm quality, quantitative abnormality, and serum T synthesis disorder caused by CP. This mechanism may be correlated with the inhibition of oxidative stress and antioxidation levels. Therefore, CGA has a protective impact on testicular injuries arising from CP in mice.

Endoplasmic reticulum stress of endometrial mesenchymal stem cells in endometriosis

ObjectiveThe human endometrium has significant regenerative abilities due to stem cells, which are vital in immunomodulation, immune tolerance, steroid hormone response, and inflammation. Endometriosis, an inflammatory gynecological disorder where endometrium-like tissue grows outside uterus, affects millions of women and often causes infertility. Recent research indicates that stem cells contribute to pathology of endometriosis. ER stress is implicated in various diseases, including endometriosis. This study aims to examine ER stress in eMSCs within endometriosis pathogenesis and uncover underlying disease mechanisms. MethodsSamples were collected from healthy subjects and women with endometriosis in both proliferative and secretory phases. eMSCs were isolated and characterized via flow cytometry. ER stress protein levels were assessed using proteomic analysis, with validation through Western Blot and immunofluorescence staining. Gene expression was analyzed by RT-qPCR, and ultrastructural examination of eMSCs was conducted using TEM. ER stress markers in tissue samples were detected in SUSD2+ eMSCs through immunofluorescence staining and visualized using a confocal microscope. Statistical analysis was performed using SPSS program. ResultsThe proteomics analysis uncovered ER stress-related proteins (DDRGK1, RTN3, ERp44, TMED2, TMEM33, TMX3) whose levels were significantly distinct from control group. Western Blot analysis and immunofluorescence staining results at protein level; RT-qPCR results at gene level supported these findings. TEM analysis also showed ultrastructural presence of ER stress in endometriosis groups. ConclusionPresence of ER stress in eMSCs in pathogenesis of endometriosis has been demonstrated using various methods. Our research has potential to shed light on pathology of endometriosis and offer promising avenues for non-invasive diagnosis and potential treatment.

Proteotoxicity and Apical Toxicity of Nicosulfuron to Danio rerio Embryos: A Comprehensive Assessment at Different Temperatures and pH

In the present study, the toxicity of nicosulfuron to Danio rerio embryos was evaluated in three experiments through standardized toxicity tests according to OECD TG236 guidelines. In the first experiment, six concentrations of nicosulfuron (0, 0.1, 1, 10, 100, 1000 mg/L) were tested under optimal conditions (26 °C, pH 7.0) to assess the general sensitivity of zebrafish embryos to nicosulfuron. The second and third experiment examined the effects of different pH levels (5.0 and 9.0) and temperatures (21 °C and 31 °C) on the toxicity at four nicosulfuron concentrations (0, 10, 100, 1000 mg/L). Additionally, the sub-organismic effects of nicosulfuron on stress protein levels (Hsp70) of fish embryos were analyzed. Throughout the embryo experiments, no malformations were observed in all experiments. The survival rate exceeded 80% in all groups except for the 21 °C (pH 7.0) treatment groups. No significant effect of nicosulfuron on the survival rate was found at the same temperature or pH (p &gt; 0.05). No significant difference in the heart rate was found among all nicosulfuron groups (p &gt; 0.05) at 21 °C. The heart rate of fish embryos at 31 °C, pH 5.0 and pH 9.0 increased with nicosulfuron concentrations. Except for the pH 5.0 (26 °C) and 21 °C (pH 7.0) treatment groups, nicosulfuron was found to increase the hatching rate of embryos in other treatments; however, the corresponding times of action were different. At 21 °C (pH 7.0), the embryos did not hatch until 144 h post-fertilization. In terms of proteotoxicity, nicosulfuron was found to be more toxic to zebrafish embryos in the 21 °C, pH 5.0 and pH 9.0 treatment groups. However, at 31 °C, no significant difference in Hsp70 levels was found among all the different nicosulfuron concentrations (p &gt; 0.05). Our results show that nicosulfuron exerts a weak toxicity to zebrafish embryos; however, this toxicity is amplified by inappropriate pH or temperature conditions.

Zinc oxide nanoparticles loaded with linalool as a potential control agent of malaria infection

This research aimed to produce and analyze zinc oxide nanoparticles (ZNPs) loaded with linalool (LZNPs), and to evaluate their in vitro and in vivo efficacy through targeting the inflammation and oxidative stress. LZNPs were synthesized using an ethanolic solution of polyvinyl alcohol. The Malstat technique was used to evaluate the effectiveness of LZNPs against both sensitive and resistant strains of Plasmosium falciparum. In vivo effects of ZNPs and LZNPs on parasite growth suppression, survival rate, oxidative stress markers, antioxidant genes, and gene and protein levels of inflammatory cytokines were evaluated by Real-time PCR and Western blot techniques. The results indicated that LZNPs demonstrated noteworthy (P < 0.001) antiplasmodial activity against both susceptible and resistant strains of P. falciparum. P. berghei NK65 strain-infected mice treated with the ZNPs and LZNPs at doses of 5–15 mg/kg notably (p < 0.001) increased the survival rates and parasite growth suppression. LZNPs at 5–15 mg/kg demonstrated a significant (p < 0.001) decrease in oxidative stress markers, increased the expression level of antioxidant genes, and reduced the gene and protein expression level of inflammatory cytokines. The current experimental study demonstrated the potent in vitro antiplasmodial activity of LZNPs against chloroquine-resistant and sensitive strains of P. falciparum compared to ZNPs alone. Additionally, the study identified the potential benefits of this nanocomposite in suppressing the parasite and extending the survival rate in mice infected with P. berghei by targeting inflammation and oxidative stress. It also showed minimal toxicity in liver and kidney function in healthy mice. Nevertheless, further research is essential to elucidate the comprehensive mechanisms and practical effectiveness of LZNPs.

Exercise mitigates reductive stress-induced cardiac remodeling in mice

The endoplasmic reticulum (ER) regulates protein folding and maintains proteostasis in cells. We observed that the ER transcriptome is impaired during chronic reductive stress (RS) in cardiomyocytes. Here, we hypothesized that a prolonged moderate treadmill exercise mitigates the RS-induced ER dysfunction and cardiac remodeling in cardiac-specific constitutively active Nrf2 mice (CaNrf2-TG). RNA sequencing showed notable alterations in the ER transcriptome of TG hearts at 4, 12, and 24 weeks (16, 28, and 35 genes, respectively). Notably, the downregulation of ER genes was significant at 12 weeks, and further pronounced at 24 weeks, at which the cardiac pathology is evident. We also observed increased levels of ubiquitinated proteins in CaNrf2-TG hearts across all ages, along with VCP, a marker of ERAD function, at 24 weeks. These findings indicate that constitutive Nrf2 activation and RS impair protein-folding activity and augments ERAD function over time. Exercise intervention for 20 weeks (beginning at 6 weeks of age), reduced cardiomyocyte hypertrophy (from 448 μm2 to 280 μm2) in TG mice, through adaptive remodeling, and preserved the cardiac function. However, while exercise did not influence antioxidants or ER stress protein levels, it significantly improved ERAD function and autophagy flux (LC-I to LC-II) in the TG-EXE hearts. Collectively, our findings underscore the prophylactic potential of exercise in mitigating RS-associated pathology, highlighting its essential role in maintaining cellular proteostasis through ER-independent mechanisms.

Ferulic Acid Inhibits Arsenic-Induced Colon Injury by Improving Intestinal Barrier Function.

The prolonged exposure to arsenic results in intestinal barrier dysfunction, which is strongly concerned with detrimental processes such as oxidative stress and the inflammatory response. Ferulic acid (FA), as a phenolic acid, possesses the capability to mitigate arsenic-induced liver damage and cardiotoxic effects dependent on inhibition of oxidative stress and inflammatory responses. FA can mitigate testicular tissue damage and alveolar epithelial dysfunction, the mechanism of which may rely on nuclear factor erythroid 2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) activation and nuclear factor-kappa B (NF-κB) pathway blocking. Based on the antioxidant and anti-inflammatory properties of FA, we speculated that FA might have the potential to inhibit arsenic-induced intestinal damage. To confirm this scientific hypothesis, mice exposed to sodium arsenite were treated with FA to observe colonic histopathology and TJ protein levels, and oxidative stress and TJ protein levels in Caco-2 cells exposed to sodium arsenite were assessed after FA intervention. In addition, molecular levels of NF-κB and Nrf2/HO-1 pathway in colon and Caco-2 cells were also detected. As shown in our data, FA inhibited arsenic-induced colon injury, which was reflected in the improvement of mucosal integrity, the decrease of down-regulated expression of tight junction (TJ) proteins (Claudin-1, Occludin, and ZO-1) and the inhibition of oxidative stress. Similarly, treatment with FA attenuated the inhibitory effect of arsenic on TJ protein expression in Caco-2 cells. In addition to suppressing the activation of NF-κB pathway, FA retrieved the activation of Nrf2/HO-1 pathway in colon and intestinal epithelial cells induced by arsenic. In summary, our findings propose that FA has the potential to mitigate arsenic-induced intestinal damage by preserving the integrity of intestinal epithelial TJs and suppressing oxidative stress. These results lay the groundwork for the potential use of FA in treating colon injuries caused by arsenic.

SERCA1 Overexpression in Skeletal Muscle Attenuates Muscle Atrophy and Improves Motor Function in a Mouse Model of ALS.

Amyotrophic lateral sclerosis (ALS) is characterized by progressive loss of muscle mass and muscle function. Previous work from our lab demonstrated that skeletal muscles from a mouse model of ALS show elevated intracellular calcium (Ca2+) levels and heightened endoplasmic reticulum (ER) stress. To investigate whether overexpression of sarcoplasmic reticulum (SR) Ca2+ ATPase 1 (SERCA1) in skeletal muscle would improve intracellular Ca2+ handling, attenuate ER stress, and improve motor function ALS transgenic mice. B6SJL-Tg (SOD1*G93A)1Gur/J (ALS-Tg) mice were bred with skeletal muscle α-actinin SERCA1 overexpressing mice to generate wild type (WT), SERCA1 overexpression (WT/+SERCA1), ALS-Tg, and SERCA1 overexpressing ALS-Tg (ALS-Tg/+SERCA1) mice. Motor function (grip test) was assessed weekly and skeletal muscles were harvested at 16 weeks of age to evaluate muscle mass, SR-Ca2+ ATPase activity, levels of SERCA1 and ER stress proteins - protein disulfide isomerase (PDI), Grp78/BiP, and C/EBP homologous protein (CHOP). Single muscle fibers were also isolated from the flexor digitorum brevis muscle to assess changes in resting and peak Fura-2 ratios. ALS-Tg/+SERCA1 mice showed improved motor function, delayed onset of disease, and improved muscle mass compared to ALS-Tg. Further, ALS-Tg/+SERCA1 mice returned levels of SERCA1 protein and SR-Ca2+ ATPase activity back to levels in WT mice. Unexpectedly, SERCA-1 overexpression increased levels of the ER stress maker Grp78/BiP in both WT and ALS-Tg mice, while not altering protein levels of PDI or CHOP. Lastly, single muscle fibers from ALS-Tg/+SERCA1 had similar resting but lower peak Fura-2 levels (at 30 Hz and 100 Hz) compared to ALS-Tg mice. These data indicate that SERCA1 overexpression attenuates the progressive loss of muscle mass and maintains motor function in ALS-Tg mice while not lowering resting Ca2+ levels or ER stress.

Hora est. The relationship between stress, salivary proteins and caries experience in children

Dental caries is a major public health problem and untreated caries has serious consequences for children. Psychosocial factors have multiple consequences, among others on the composition of saliva. Therefore, this study investigated whether stress and various salivary protein levels are associated with dental caries experience in children. The activity of the Matrix Metalloproteinases MMP-8 and MMP-9 and the total proteolytic activity in saliva turned out to be indicators for the caries experience. Salivary Alpha-Amylase seems to be a protector and was a strong indicator for caries experience. In cases where children were exposed to two different dental treatments, the level of salivary cortisol- and alpha-amylase increased, in which a distinction could be made between non-invasive and invasive treatment. The results of the study emphasise the need for further research into the way stress and salivary protein concentrations can affect the caries experience and how different dental treatments can influence the behaviour and stress levels in children.

Combination of Polygonatum Rhizoma and Scutellaria baicalensis triggers apoptosis through downregulation of PON3-induced mitochondrial damage and endoplasmic reticulum stress in A549 cells.

Scutellaria baicalensis (SB) and Polygonatum Rhizoma (PR), two traditional Chinese medicines, are both known to suppress cancer. However, the mechanism and effect of combined treatment of them for lung cancer are rarely known. Investigating the combined effect of SB and PR(hereafter referred to as SP) in potential mechanism of lung cancer is required. This study was to evaluate the inhibitory effects of SP on A549 cell growth and to explore the underlying molecular mechanisms. According to the theory of Chinese medicine and network pharmacology, in the in vivo experiment, a mouse model of carcinoma in situ was constructed, and lung carcinoma in situ tissues were collected for proteomics analysis, hematoxylin-eosin staining, and CK19 immunohistochemistry. In the in vitro experiment, lung cancer A549 cells at logarithmic growth stage were taken, and the inhibitory effect of SP on the proliferation of A549 cells was detected by CCK8 method. The expression of PON3 was detected by quantitative polymerase chain reaction and western blot. In addition, the effect of SP on the induction of apoptosis in A549 cells and the changes of membrane potential and reactive oxygen species (ROS) content were detected by flow cytometry. The changes of PON3 content in endoplasmic reticulum (ER) are observed by laser confocal microscopy, whereas the effects of SP on the expression of apoptosis-related proteins and ER stress-related proteins in A549 cells were examined by western blot. By searching the Traditional Chinese Medicines of Systems Pharmacology (TCMSP) (https://www.tcmspe.com/index.php) database and SymMapdatabase, the respective target genes of PR and SB were mapped into protein network interactions, and using Venn diagrams to show 38 genes in common between PR and SB and lung cancer, SPwas found to play a role in the treatment of lung cancer. In vivo experiments showed that in a lung carcinoma in situ model, lung tumor tissue was significantly lower in the SP group compared with the control group, and PON3 was shown to be downregulated by lung tissue proteomics analysis. The combination of SP was able to inhibit the proliferation of A549 cells in a concentration-dependent manner (p < .0001). The expression levels of apoptosis-related proteins and ER stress proteins were significantly increased and the expression levels of PON3 and anti-apoptosis-related proteins were decreased in A549 cells. At the same time, knockdown of PON3 could inhibit tumor cell proliferation (p < .0001). The combination of different concentrations of SP significantly induced apoptosis in A549 cells (p < .05; p < .0001), increased ROS content (p < .01), and damaged mitochondrial membrane potential of A549 cells (p < .05; p < .0001), and significantly increased the expression levels of apoptosis-related proteins and ER stress proteins in lung cancer A549 cells. SP inhibits proliferation of lung cancer A549 cells by downregulating PON3-induced apoptosis in the mitochondrial and ER pathways.

Sustainable Release Selenium Laden with SiO2 Restoring Peripheral Nerve Injury via Modulating PI3K/AKT Pathway Signaling Pathway.

Inhibiting ROS overproduction is considered a very effective strategy for the treatment of peripheral nerve injuries, and Se has a remarkable antioxidant effect; however, since the difference between the effective concentration of Se and the toxic dose is not large, we synthesized a nanomaterial that can release Se slowly so that it can be used more effectively. Se@SiO2 NPs were synthesized using a mixture of Cu2-x Se nanocrystals, and the mechanism of action of Se@SiO2 NPs was initially explored by performing sequencing, immunofluorescence staining and Western blotting of cellular experiments. The mechanism of action of Se@SiO2 NPs was further determined by performing behavioral assays after animal experiments and by sampling the material for histological staining, immunofluorescence staining, and ELISA. The effects, mechanisms and biocompatibility of Se@SiO2 NPs for peripheral nerve regeneration were determined. Porous Se@SiO2 was successfully synthesized, had good particle properties, and could release Se slowly. CCK-8 experiments revealed that the optimal experimental doses were 100 μM H2O2 and 200 μg/mL Se@SiO2, and RNA-seq revealed that porous Se@SiO2 was associated with cell proliferation, apoptosis, and the PI3K/AKT pathway. WB showed that porous Se@SiO2 could increase the expression of cell proliferation antigens (PCNA and S100) and antiapoptotic proteins (Bcl-2), decrease the expression of proapoptotic proteins (Bax), and increase the expression of antioxidative stress proteins (Nrf2, HO-1, and SOD2). EdU cell proliferation and ROS fluorescence assays showed that porous Se@SiO2 promoted cell proliferation and reduced ROS levels. The therapeutic effect of LY294002 (a PI3K/AKT pathway inhibitor) was decreased significantly and its effect was lost when it was added simultaneously with porous Se@SiO2. Animal experiments revealed that the regenerated nerve fiber density, myelin thickness, axon area, gastrocnemius muscle wet-to-weight ratio, myofiber area, sciatic nerve function index (SFI), CMAP, apoptotic cell ratio, and levels of antioxidative stress proteins and anti-inflammatory factors were increased following the administration of porous Se@SiO2. The levels of oxidative stress proteins and anti-inflammatory factors were significantly greater in the Se@SiO2 group than in the PNI group, and the effect of LY294002 was decreased significantly and was lost when it was added simultaneously with porous Se@SiO2. Se@SiO2 NPs are promising, economical and effective Se-releasing nanomaterials that can effectively reduce ROS production, inhibit apoptosis and promote cell proliferation after nerve injury via the PI3K/AKT pathway, ultimately accelerating nerve regeneration. These findings could be used to design new, promising drugs for the treatment of peripheral nerve injury.

Zhi Zhu Ma Ren pill relieves constipation in mice through endoplasmic reticulum stress-mediated apoptosis.

This study explores the detailed effects and mechanisms of Zhi Zhu Ma Ren Pill (ZZMRP) on constipation. Mouse constipation was induced by using loperamide (Lop). The effects and mechanisms of ZZMRP on constipation were addressed by various methods including charcoal meals, hematoxylin and eosin (H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labelling (TUNEL), transmission electron microscopy, and western blot experiments. Lop-treated mice exhibited delayed transit and reduced ink progradation rates after charcoal meal administration. H&E staining confirmed severe pathologic symptoms in these mice. Additionally, a decline in interstitial Cajal cells (ICCs) was observed in Lop-treated mice, accompanied by reduced concentrations of 5-hydroxytryptamine (5-HT), acetylcholinesterase (AchE), substance P (SP), and vasoactive intestinal peptide (VIP), coupled with an elevated concentration of nitric oxide synthase (NOS). However, ZZMRP treatment markedly ameliorated these changes. In addition, ZZMRP introduction significantly reversed the Lop-induced enhancement in apoptosis rate, endoplasmic reticulum (ER) stress, and levels of proapoptotic proteins and ER stress proteins, and the decrease in the expression of antiapoptotic proteins. These effects were further confirmed with the use of 4-phenylbutyrate (4-PBA), which also reversed the changes in apoptosis rate and protein levels. ZZMRP alleviates constipation primarily through modulating ER stress-mediated apoptosis in Lop-treated mice, suggesting its use as a therapeutic agent for constipation.

LncRNA Peg13 Alleviates Myocardial Infarction/Reperfusion Injury through Regulating MiR-34a/Sirt1-Mediated Endoplasmic Reticulum Stress.

Myocardial infarction/reperfusion (I/R) injury significantly impacts the health of older individuals. We confirmed that the level of lncRNA Peg13 was downregulated in I/R injury. However, the detailed function of Peg13 in myocardial I/R injury has not yet been explored.To detect the function of Peg13, in vivo model of I/R injury was constructed. RT-qPCR was employed to investigate RNA levels, and Western blotting was performed to assess levels of endoplasmic reticulum stress and apoptosis-associated proteins. EdU staining was confirmed to assess the cell proliferation.I/R therapy dramatically produced myocardial injury, increased the infarct area, and decreased the amount of Peg13 in myocardial tissues of mice. In addition, hypoxia/reoxygenation (H/R) notably induced the apoptosis and promoted the endoplasmic reticulum (ER) stress of HL-1 cells, while overexpression of Peg13 reversed these phenomena. Additionally, Peg13 may increase the level of Sirt1 through binding to miR-34a. Upregulation of Peg13 reversed H/R-induced ER stress via regulation of miR-34a/Sirt1 axis.LncRNA Peg13 reduces ER stress in myocardial infarction/reperfusion injury through mediation of miR-34a/Sirt1 axis. Hence, our research might shed new lights on developing new strategies for the treatment of myocardial I/R injury.

γ-Aminobutyric Acid Effectively Modulate Growth Performance, Physiological Response of Largemouth Bass (Micropterus Salmoides) Under Combined Stress of Flow Velocity and Density.

The circular aquaculture model of largemouth bass pond engineering has the characteristics of high yield and efficiency, but it is prone to stress caused by flow velocity and density, which affects the yield of largemouth bass (Micropterus salmoides). γ-Aminobutyric acid (GABA) is believed to have the effect of improving growth and stress tolerance. We divided the largemouth bass into three groups: a control group, a flow rate and density combined stress group, and a combined stress feed supplemented with GABA (0.9%) group, and conducted a 60-day aquaculture experiment. The results showed that the final weight, weight gain rate (WGR), specific growth rate (SGR), and feed efficiency (FE) of largemouth bass significantly decreased in the combined stress group (P < 0.05). The serum aspartate aminotransferase (AST), alanine transaminase (ALT) activity, and glucose (GLU), malondialdehyde (MDA) level of largemouth bass significantly higher than the control group, and the serum lysozyme (LZM) activity and total antioxidant capacity (T-AOC) were significantly lower than the control group (P < 0.05). After adding GABA, the final weight, WGR, SGR, and FE decreased, and the serum GLU levels, AST, ALT activity, and MDA levels were downregulated, and the serum LZM activity and T-AOC of largemouth bass were upregulated. But most of the above are still at the level of the control group. Under combined stress, the messenger ribonucleic acid (mRNA) expression levels of growth hormone (GH), insulin-like growth factor-Ⅰ (IGF-I), B-cell lymphoma-2 (Blc2), nuclear transcription factor E2-related factor 2 (Nrf2), catalase (CAT), and superoxide dismutase (SOD) genes were significantly reduced (P < 0.05), while the mRNA expression levels of heat stress protein 70 (HSP70), heat stress protein 90 (HSP90), nuclear factor kappa-B (NF-κB), interleukin-1β (IL-1β), Bax and keap1 genes were significantly increased (P < 0.05). After the exogenous addition of GABA, all the above genes have a certain degree of callback, but GH, HSP70, HSP90, IL-1β, Bax, Nrf2, CAT, and SOD have not yet reached the level of the control group. These results indicate that adding GABA to feed can alleviate the adverse effects of combined stress of flow rate and density to a certain extent and provide insights for solving the problems in the circular aquaculture model of largemouth bass.

Anticancer and antimicrobial potential of zinc/sodium alginate/polyethylene glycol/d-pinitol nanocomposites against osteosarcoma MG-63 cells

Abstract Background The field of nanomedicine has attracted much interest and is now serving as the impetus for many revolutionary advances in the pharmaceutical industry. Objectives In the current exploration, we intended to fabricate the zinc/sodium alginate/polyethylene glycol/d-pinitol nanocomposites (ZSP/D-Pin/NCs) and evaluate their antimicrobial and anticancer properties against MG-63 cells. Methods ZSP/D-Pin/NCs were synthesized and characterized using several techniques and their cytotoxicity was examined against osteosarcoma MG-63 cells and normal 3T3 cells using the MTT assay. The levels of oxidative stress and apoptotic protein were examined using assay kits and fluorescence staining. Results and Conclusion The findings of several characterization studies revealed the development of agglomerated and crystalline ZSP/D-Pin/NCs. The antimicrobial assay demonstrated that ZSP/D-Pin/NCs substantially inhibited the growth of pathogens. Additionally, the MG-63 cell viability, which was exposed to several doses (1–20 µg) of ZSP/D-Pin/NCs, showed a remarkable decrease at various time periods, i.e., 24, 48, and 72 h without showing toxicity in 3T3 cells. The results of the fluorescence staining assay demonstrated that ZSP/D-Pin/NCs considerably increased apoptosis in the MG-63 cells by triggering oxidative stress. The antioxidants were reduced and upregulated the Bax and caspase expressions in ZSP/D-Pin/NC-treated MG-63 cells.

Danlou tablet attenuates ischemic stroke injury and blood‒brain barrier damage by inhibiting ferroptosis

Ethnopharmacological relevanceDanlou tablet (DLT) is a traditional Chinese medicinal formulation known for replenishing Qi, promoting blood circulation, and resolving stasis. Its pharmacological actions primarily involve anti-inflammatory, antioxidant stress reduction, antiapoptotic, proangiogenic, and improved energy metabolism. DLT has been confirmed to have favorable therapeutic effects on ischemic stroke (IS). However, the underlying mechanism through which DLT affects IS-induced brain injury remains unknown. Aim of the studyThis study aims to investigate the effects and underlying mechanisms of danlou tablet on ischemic stroke based on network pharmacology and experimental verification. Materials and methodsUsing a transient middle cerebral artery occlusion (tMCAO) mouse model, the impact of DLT on the blood‒brain barrier (BBB) and brain injury in mice was assessed. Network pharmacology and bioinformatics analyses were utilized to explore the potential mechanisms of DLT in treating IS. Endothelial cells were cultured to observe the effects of DLT on vascular endothelial cells after oxygen-glucose deprivation/reperfusion, and these findings were validated in the brains of tMCAO mice. ResultsDLT alleviated oxidative stress and brain damage in tMCAO mice, mitigating BBB damage. A total of 185 potential targets through which DLT regulates IS were identified, including COX2, a known critical marker of ferroptosis, which identified as a key target. In vitro and in vivo experiments demonstrated that DLT significantly (p < 0.05) improved cell death and vascular barrier damage in IS, reducing intracellular oxidative stress and COX2 protein levels while increasing SLC7A11 and GPX4 protein levels. ConclusionsThis study demonstrated that DLT maintained BBB integrity and alleviated brain injury of tMCAO mice by inhibiting ferroptosis. The study partially unraveled the mechanism through which DLT functioned in treating IS and further clarified the pivotal active components of DLT, thereby providing a theoretical scientific basis for treating IS with DLT.

Proteomic insight into human directed selection of the domesticated chicken Gallus gallus.

Chicken domestication began at least 3,500 years ago for purposes of divination, cockfighting, and food. Prior to industrial scale chicken production, domestication selected larger birds with increased egg production. In the mid-20th century companies began intensive selection with the broiler (meat) industry focusing on improved feed conversion, rapid growth, and breast muscle yield. Here we present proteomic analysis comparing the modern broiler line, Ross 708, with the UIUC legacy line which is not selected for growth traits. Breast muscle proteome analysis identifies cellular processes that have responded to human directed artificial selection. Mass spectrometry was used to identify protein level differences in the breast muscle of 6-day old chicks from Modern and Legacy lines. Our results indicate elevated levels of stress proteins, ribosomal proteins and proteins that participate in the innate immune pathway in the Modern chickens. Furthermore, the comparative analyses indicated expression differences for proteins involved in multiple biochemical pathways. In particular, the Modern line had elevated levels of proteins affecting the pentose phosphate pathway, TCA cycle and fatty acid oxidation while proteins involved in the first phase of glycolysis were reduced compared to the Legacy line. These analyses provide hypotheses linking the morphometric changes driven by human directed selection to biochemical pathways. These results also have implications for the poultry industry, specifically Wooden Breast disease which is linked to rapid breast muscle growth.