Heat Stress-induced Damage Research Articles

Melatonin alleviates heat stress-induced testicular damage in dairy goats by inhibiting the PI3K/AKT signaling pathway

Current measures mainly focus on how melatonin reduces physiological heat stress in animals, but its effects on reproductive damage to male dairy goats have been neglected. This study aimed to determine the protective effect of melatonin on male reproduction during heat stress in dairy goats and to further explore its mechanisms. A natural heat stress model of Saanen dairy goats was used to assess testicular tissue damage 7 days after heat stress and to examine semen quality changes during a spermatogenic cycle. RNA-seq, Western blot, RT–qPCR, and immunofluorescence staining were used to explore the mechanism by which melatonin protects against heat stress-induced reproductive damage and to validate the results. The data suggested that melatonin significantly alleviated the heat stress-induced decrease in sperm quality, protected varicose tubule structure, reduced the levels of heat shock proteins and apoptotic proteins and protected the spermatocytes and round spermatozoa, which are mainly affected by heat stress. RNA-seq results suggest that melatonin inhibits the PI3K/AKT signaling pathway, reduces the level of p-AKT, and promotes elevated BCL-2. In addition, melatonin treatment could upregulate the gene expression of MT2 which was downregulated by heat stress and improve the change in extracellular matrix components and restore serum testosterone levels. Our results suggest that melatonin can protect against testicular and spermatogenic cell damage and improve semen quality in male dairy goats under heat stress. This study provides an important reference for subsequent studies on the molecular mechanisms of melatonin in protecting male reproductive processes under heat stress and using exogenous melatonin to prevent heat stress.

Effects of N-acetyl-l-cysteine on chronic heat stress-induced oxidative stress and inflammation in the ovaries of growing pullets

The aims of this study were to investigate the effects of supplemental N-acetyl-l-cysteine (NAC) on chronic heat stress-induced oxidative stress and inflammation in the ovaries of growing pullets. A total of 120, 12-wk-old, Hy-Line Brown hens were randomly separated into 4 groups with 6 replicates of 5 birds in each group for 21 d. The 4 treatments were as follows: the CON group and CN group were supplemented with basal diet or basal diet with 1 g/kg NAC, respectively; and the HS group and HSN group were heat-stressed groups supplemented with basal diet or basal diet with 1 g/kg NAC, respectively. The results indicated that the ovaries suffered pathological damage due to chronic heat stress and that NAC effectively ameliorated these changes. Compared with the HS group, antioxidant enzyme activities (including SOD, GSH-Px, CAT, and T-AOC) were enhanced, while the MDA contents and the expression levels of HSP70 were decreased in the HSN group. In addition, NAC upregulated the expression levels of HO-1, SOD2, and GST by upregulating the activity of Nrf2 at different time points to mitigate oxidative stress caused by heat exposure. Simultaneously, NAC attenuated chronic heat stress-induced NF-κB pathway activation and decreased the expression levels of the proinflammatory cytokines IL-8, IL-18, TNF-α, IKK-α, and IFN-γ. Cumulatively, our results indicated that NAC could ameliorate chronic heat stress-induced ovarian damage by upregulating the antioxidative capacity and reducing the secretion of proinflammatory cytokines.

Inoculation with Arbuscular Mycorrhizal Fungi Alleviates the Adverse Effects of High Temperature in Soybean.

High temperature is foremost abiotic stress and there are inadequate studies explicating its impact on soybean. In this study, a pot experiment was done in a greenhouse maintained at a day/night temperature of 42/28 °C with a mean temperature of 35 °C to examine the effects of high temperature in soybean plants inoculated with and without arbuscular mycorrhizal fungi (AMF).Various parameters were taken in soybean plants treated with AMF (+) and AMF (−) such as growth analysis, chlorophyll content, canopy temperature, number of stomata, gas exchange, chlorophyll fluorescence, seed yield, and its attributes. It was observed that growth parameters like leaf area, stem height, root length, shoot and root dry biomass were increased in AMF (+) as compared to AMF (−) plants. Chlorophyll content, the number of stomata, photosynthesis rate, stomatal conductance, transpiration rate, and water use efficiency increased in AMF (+) as compared to AMF (−) plants. Chlorophyll fluorescence parameters such as Fv/Fm, Fv/Fo, PhiPSII, fluorescence area, performance index, photochemical quenching, linear electron transport rate, and active reaction centres density of PSII were also found to be enhanced in AMF (+) plants. However, canopy temperature, intercellular CO2, Fo/Fm, and non-photochemical quenching were higher in AMF (−) as compared to inoculated plants. An increase in growth and photosynthesis ultimately enhanced the seed yield and its attributes in AMF (+) as compared to AMF (−). Thus, AMF (+) plants have shown much better plant growth, photosynthesis parameters, and seed yield as compared to AMF (−) plants under high temperature. Thus, it is concluded that heat stress-induced damage to the structure and function of the photosynthetic apparatus was alleviated by AMF inoculum. Therefore, AMF can be used as a biofertilizer in alleviating the adverse effects of heat stress in soybean.

Depletion of serum-derived exosomes aggravates heat stress-induced damage of bovine mammary epithelial cells.

Exosomes are involved in intercellular communication, affecting many physiological and pathological process. The present study evaluated the effects of serum exosomes on the function of bovine mammary epithelial cells (BMECs) and milk synthesis under heat stress. We cultured the BMECs in fetal bovine serum (FBS) or exosome-free FBS medium and examined, their viability using CCK-8 kit. The results showed that culturing the cells in an exosome-free medium decreased viability and increased the levels of reactive oxygen species. The BMECs cultured in the exosome-free medium had reduced mitochondrial membrane potential, decreased manganese superoxide dismutase activity, and disrupted mitochondrial dynamics. They exhibited apoptosis due to upregulated Drp1, Fis1, Bax and HSP70. Lastly, we observed downregulation of milk fat and lactoprotein-related genes: mTOR, PPARγ, p-mTOR and ADD1 and SREBP1, ELF5, and CSN2, respectively, after culturing the cells in an exosome-free medium. These negative effects of the exosome-free medium on the BMECs could be further reinforced under heat stress. Our results demonstrated that exosomes from serum are critical for maintaining the normal function of BMECs.

Linoleic and linolenic acids reduce the effects of heat stress-induced damage in pig oocytes during maturation in vitro.

Elevated environmental temperatures can induce heat stress which could reduce fertility and early embryonic development. Fatty acids can initiate an endergonic reaction that absorbs cellular heat and decreases intracellular temperature. This study's objective was to minimize heat stress-induced damage to in vitro matured oocytes by supplementing maturation media with either 50μM linoleic or linolenic acid or both (25 or 50μM) during maturation at either 38.5 or 41.5°C. Oocytes were evaluated for intracellular antioxidative pathways, fertilization characteristics, or early embryonic development. Elevated maturation temperatures increased (p < 0.05) reactive oxygen species (ROS) formation and supplementation with linoleic or linolenic acid decreased (p < 0.05) ROS in oocytes matured at 41.5°C. Maturation temperature had an effect (p < 0.05) on the intracellular antioxidative pathways of the oocyte except for glutathione peroxidase activity. Regardless of maturation temperature, supplementation with linoleic or linolenic acid increased (p < 0.05) the enzyme activities and glutathione concentrations in the oocytes compared to no fatty acid supplementation. Supplementation of both linoleic and linolenic acid decreased (p < 0.05) polyspermic fertilization rates. Supplementing either 25 or 50μM linoleic and linolenic acid to maturing oocytes at 41.5°C increased (p < 0.05) cleavage rates by 48h after IVF and the blastocyst formation rates by 144h after IVF compared to other treatments. Oocytes matured at 38.5°C had greater (p < 0.05) embryonic development than those matured at 41.5°C except for those supplemented with 50μM linoleic and linolenic acid. Supplementing 50μM linoleic and linolenic acid to the maturation medium of pig oocytes reduces the effects of heat stress-induced damage.

Chronic heat stress causes liver damage via endoplasmic reticulum stress-induced apoptosis in broilers

Liver is a central metabolic organ, which is sensitive to heat stress. Liver damage affects animals' health and endangers the livestock and poultry industry. This study aimed to investigate the mechanism of chronic heat stress-induced liver damage in broiler chickens. Broilers were divided into 3 treatments: normal control group (NOR, 22°C), heat stress group (HS, 32°C) and pair-feeding group (PF, 22°C) for a 7-d and 14-d trial. The results showed that 7 d heat exposure caused microvesicular steatosis and reduced glutamine synthetase activity in broiler liver (P < 0.05). After 14 d of heat exposure, heat stress caused vacuolar degeneration and apoptosis in the liver; elevated liver relative weight and liver glutaminase activity as well as plasma ammonia level (P < 0.05). Additionally, heat stress enhanced GRP78 protein expression and the mRNA expressions of endoplasmic reticulum (ER) stress responses genes and apoptosis-related genes in broiler liver after 14 d of heat exposure (P < 0.05). In conclusion, chronic heat stress triggered ER stress-induced apoptosis and caused liver damage, which may compromise ammonia detoxification in broiler liver.

Heat acclimation ameliorated heat stress-induced acute kidney injury and prevented changes in kidney macrophages and fibrosis.

Heatstroke can cause acute kidney injury (AKI), which reportedly progresses to chronic kidney disease. Kidney macrophages may be involved in such injury. Although heat acclimation (HA) provides thermal resilience, its renoprotective effect and mechanism remain unclear. To investigate heat stress-induced kidney injuries in mice and the mitigating effect of HA on them, male C57/BL6J mice were exposed to heat stress (40°C, 1 h) with or without 5-day HA (38°C, 3 h/day) prior to heat stress. Heat stress damaged kidney proximal tubules with an elevation of urinary kidney injury molecule-1. Kidney fibrosis was observed on day 7 and correlated with urinary kidney injury molecule-1 levels on day 3. Kidney resident macrophages decreased on day 1, whereas the number of infiltrating macrophages in the kidney did not change. Both subsets of macrophages polarized to the proinflammatory M1 phenotype on day 1; however, they polarized to the anti-inflammatory M2 phenotype on day 7. HA significantly ameliorated heat stress-induced proximal tubular damage and kidney fibrosis. HA substantially increased heat shock protein 70 expression in the tubules before heat stress and reduced the elevation of cleaved caspase-3 expression after heat stress. HA also induced heat shock protein 70 expression of resident macrophages and prevented heat stress-induced changes in both subsets of kidney macrophages. These results provide pathophysiological data supporting the renoprotective effect of HA. Further studies are needed to confirm that HA can prevent kidney damage due to heat stress in humans.NEW & NOTEWORTHY Heat stress could induce acute kidney injury. Although heat acclimation (HA) reportedly provides thermal tolerance, its effect on heat stress-induced kidney damage remains unclear. This study showed that 5-day HA ameliorates mouse kidney tubular damage and subsequent fibrosis caused by heat stress. It also demonstrated that HA enhances intracellular heat shock protein 70 expression in tubular cells and prevents a decrease in kidney resident macrophages, which explains the renoprotective effect of HA.

Protective effects of different concentrations of selenium nanoparticles on rainbow trout (Oncorhynchus mykiss) primary hepatocytes under heat stress

Heat stress leads to altered expression of associated heat shock proteins (HSPs), which are critical molecular chaperones related to cellular function in living organisms. Selenium nanoparticles (SeNPs), a nanocomposite form of Se, have a protective effect against heat stress-induced cellular damage. In this study, primary rainbow trout hepatocytes were isolated to identify the protective function of SeNPs in rainbow trout hepatocytes. Experiments were divided into five groups and SeNPs were added at concentrations of 0, 2.0, 3.0, 5.0 and 8.0 μg/mL and incubated at 18 ℃ for 4, 8, 12, 24 and 48 h respectively. Hepatocyte viability, GSH-Px and SOD activity were enhanced and MDA content was reduced following the addition of SeNPs. Expression of GSH-P1 and genes related to HSPs (including HSP70a, HSP60, HSP90β, HSP10 and HSP47) were significantly increased and the optimal concentration of SeNPs for adding to hepatocytes was identified as 5.0 µg/mL. Adding 5.0 µg/mL SeNPs following heat stress (24 ℃) increased hepatocyte viability, GSH-Px and SOD activity, while MDA levels first decreased and then increased. Expression of GSH-P1 and genes related to HSPs (including HSP70a, HSP60, HSP90β, HSP10 and HSP47) were significantly higher than controls. In summary, SeNPs and slight heat stress synergistically enhanced the expression of GSH-P1 and HSPs and protected hepatocytes from heat stress damage, suggesting that SeNPs is a potential hepatocyte protective therapeutic agent.

Extranuclear p53 suppresses autophagy through AMPK/mTOR signaling to promote heat stress-induced vascular endothelial cell damage

To explore the role of extranuclear p53-mediated autophagy suppression by regulating AMPK/mTOR signaling pathway in heat stress (HS)-induced injury of mouse aortic endothelial cells (MAECs). Primary cultures of MAECs were pretreated with compound C (an AMPK inhibitor), rapamycin (a mTOR inhibitor) or pifithrin-α (PFT, a selective p53 inhibitor) for 1 h before exposure to HS (43 ℃) for 2 h. The changes in cell viability at different time points after HS were examined using CCK-8 assay, and the protein expressions of P53, LC3-II, Beclin-1, p62 and the AMPK/mTOR signaling proteins were detected using Western blotting. In the animal experiment, C57 mice were pretreated with compound C, rapamycin or PFT and exposed to a high temperature at 40 ℃ to induce HS. The pathological changes in the aorta of the mice were observed with HE staining, and cell apoptosis was detected using TUNEL staining. In cultured MAECs, the cell viability was significantly reduced (P < 0.05) and the mitochondrial fraction of p53 increased while its cytoplasmic fraction decreased progressively over time following HS. HS significantly lowered the expressions of LC3-II and Beclin-1, increased p62 level, suppressed AMPK phosphorylation, and increased mTOR phosphorylation and the expressions of its downstream proteins at 6 h after the exposure (P < 0.05). Pretreatment with compound C significantly inhibited LC3-II and Beclin- 1 expression, enhanced p62 expression, and aggravated HS-induced cell injury and apoptosis in MAECs; rapamycin treatment produced the opposite effects (P < 0.05). PFT treatment significantly enhanced the viability of MAECs and alleviated HSinduced injury and apoptosis; PFT also significantly promoted activation of AMPK phosphorylation, inhibited mTOR phosphorylation and its downstream proteins (P < 0.05), enhanced the expressions of LC3-II and Beclin 1, and inhibited p62 expression in the MAECs (P < 0.05). In C57 mice, HS resulted in swelling, shedding and apoptosis of aortic vascular endothelial cells. Pretreatment with compound C obviously aggravated HS-induced vascular injury and endothelial cell apoptosis, while pretreatment with either rapamycin or PFT significantly alleviated these injuries. Autophagy inhibition mediated by extranuclear p53 via inhibiting AMPK activity and activating mTOR signaling participates in HS-induced injury of MAECs.

Nrf2 activation mediates the protection of mouse Sertoli Cells damage under acute heat stress conditions

Heat stress is known to negatively impact the reproductive process of livestock, which inevitably leads to a decline in animal fertility. Nuclear factor E2-related factor 2 (Nrf2) is an inducible transcription factor, which is essential for maintaining redox signal transmission against oxidative stress. However, there is no reliable research on the response mechanism of Sertoli Cells (SCs) against heat stress and the activation of Nrf2 when SCs are exposed to heat stress. Here, we used primary mouse SCs and SCs line TM4, along with Nrf2 specific inhibitor to determine the reaction mechanism of SCs to maintain intracellular redox homeostasis and self-survival by activating Nrf2. We found that acute heat stress only affected the vitality of SCs and the expression of functional molecules (tight junction-associated proteins and lactate dehydrogenase A [LDHA]) but did not cause cell apoptosis. When Nrf2 was inhibited, more cell death occurred in TM4 cells post heat stress treatment, along with a greater decrease in cell viability and a significant increase in intracellular ROS levels. Our study clarified for the first time the protective effect of Nrf2 activation on heat stress-induced SCs damage. It explained the possible reasons or mechanisms involved in the survival of SCs, the critical protective cells in the testis, which were not affected by heat stress. This study further improved the response mechanism of SCs in the reproductive injury caused by a high-temperature environment.

Cold-inducible RNA-binding protein regulates cyclin B1 against spermatogenesis arrest caused by heat stress.

Spermatogenesis arrest and spermatogenic cell apoptosis occur in the testes of heat-stressed mice. Although heat stress-induced spermatogenic cell apoptosis is due to the decreased expression of cold-inducible RNA-binding protein (CIRBP), it remains unclear whether spermatogenesis arrest is also affected by CIRBP. Additionally, the specific mechanism by which CIRBP regulates spermatogenic cell apoptosis or inhibits spermatogenesis remains to be elucidated. To investigate the mechanism by which CIRBP contributes to heat stress-induced testicular spermatogenesis arrest. Target mRNAs downstream of CIRBP in testicular tissue of BALB/c mice, exposed or not to heat stress, were sequenced. Sequencing data were subjected to bioinformatics analysis to identify key mRNAs and pathways associated with heat stress-induced spermatogenic damage. The link between CIRBP and its target mRNA Ccnb1 (cyclin B1) was verified by western blotting, flow cytometry, and RNA pulldown assays, and the ability of CIRBP to inhibit germ cell cycle arrest by regulating cyclin B1 expression was investigated in a mouse spermatocyte cell line (GC-2spd). Changes in mRNA expression downstream of CIRBP were mainly associated with the cell cycle and RNA binding, transport and splicing. Cyclin B1 was found to regulate the G2/M transition during the first meiotic division of spermatogenic cells. Further, CIRBP was shown to bind directly to the 3'-untranslated region of Ccnb1 mRNA and was associated with cyclin B1-induced inhibition of spermatogenesis arrest. In conclusion, our results provide strong evidence that CIRBP may exert its key function in heat stress-induced testicular spermatogenic cell injury partly by regulating the expression of Ccnb1, the product of which inhibits spermatogenesis arrest.

Vitamin A alleviates heat stress-induced damage to hair follicle development in Rex rabbits.

Rex rabbits are important fur rabbits. Heat stress severely reduces the fur quality of Rex rabbits. The aim of this study was to experimentally investigate the effect of dietary vitamin A (VA) addition on hair follicle development and related signal pathways in Rex rabbits under heat stress. In the experiment, 90 Rex rabbits were randomly divided into three groups: control group (20-25 °C, fed basic diet), heat stress group (30-34 °C, fed basic diet), and heat stress + VA group (20-25 °C, fed 12 000 IU/kg VA in addition to the basic diet). VA could significantly increase the hair follicle density (P < 0.01), hair length (P < 0.05), and the ratio of secondary to primary hair follicles (P < 0.05). In addition, VA could significantly inhibit the expression of BMP2, BMP4, FGF5, TGF-β1, and miR-214 in heat-stressed Rex rabbits and significantly increase the expression of noggin, IGF1, IGF1R, Wnt10b, CTNNB1, SHH, and miR-203 and the levels of Wnt10b and p-β-catenin; however, there was no significant effect of VA on the expression of EGF and miR-205. The dietary addition of VA can increase the hair follicle density and fur quality of heat-stressed Rex rabbits. Wnt10/β-catenin, insulin-like growth factor 1 (IGF1), fibroblast growth factor 5 (FGF5), noggin-BMP, and sonic hedgehog (SHH) signaling were associated with VA regulation under heat stress. It is possible that miR-205 and miR-194 contribute to the regulation of Wnt10/β-catenin and bone morphogenetic protein (BMP) signaling. © 2021 Society of Chemical Industry.

Spermidine Induces Expression of Stress Associated Proteins (SAPs) Genes and Protects Rice Seed from Heat Stress-Induced Damage during Grain-Filling.

Heat stress during seed maturation significantly reduced seed size and quality. Polyamines, especially spermidine (Spd), were reported to be closely related to seed development and plant heat tolerance. Stress-associated proteins (SAPs) also played a critical role in plant heat resistance, but the relationship between Spd and SAPs in improving rice tolerance to heat stress during grain filling has not been reported. Our results showed that the external spraying Spd (1.5 mM) significantly increased seed germination rate, germination index, vigor index and 1000-grain weight, significantly increased endogenous Spd, spermine (Spm) content and peroxidase activity; significantly reduced MDA content; and greatly alleviated the impact of heat stress on rice seed quality during grain filling stage as compared with high temperature control. OsSAP5 was the most upregulated expression induced by Spd, and may be mainly involved in the Spd-mediated enhancement of high-temperature resistance during rice seed development. Overexpression of OsSAP5 in Arabidopsis enhanced 1000-grain weight and seed heat resistance. Exogenous Spd alleviated the survival rate and seedling length, reduced MDA content, and upregulated the expression levels of SPDS and SPMS in Atsap4 mutant under high temperature during seed germination. In all, exogenous Spd alleviated the heat damage on seed quality during the grain filling stage and seed germination stage by improving endogenous Spd and Spm. OsSAP5, a key gene induced by Spd, might be involved in the rice heat resistance and seed quality in coordination with Spd and Spm.

Heat Stress Alters the Intestinal Microbiota and Metabolomic Profiles in Mice.

BackgroundHeat stress has negative effects on the intestinal health of humans and animals. However, the impact of heat stress on intestinal microbial and metabolic changes remains elusive. Here, we investigated the cecal microbial and metabolic profiles in mice in response to heat stress.MethodsThe mouse heat stress model was constructed by simulating a high-temperature environment. Twenty mice were randomly assigned to two groups, the control group (CON, 25°C) and the heat treatment group (HS, 40°C from 13:00 to 15:00 every day for 7 days). Serum and cecal contents were collected from the mice for serum biochemical analysis, 16S rRNA high-throughput sequencing, and non-targeted metabolomics.ResultsBoth core body temperature and water intake were significantly increased in the HS group. Serum biochemical indicators were also affected, including significantly increased triglyceride and decreased low-density lipoprotein in the heat stress group. The composition and structure of intestinal microbiota were remarkably altered in the HS group. At the species level, the relative abundance of Candidatus Arthromitus sp. SFB-mouse-Japan and Lactobacillus murinus significantly reduced, while that of Lachnospiraceae bacterium 3-1 obviously increased after HS. Metabolomic analysis of the cecal contents clearly distinguished metabolite changes between the groups. The significantly different metabolites identified were mainly involved in the fatty acid synthesis, purine metabolism, fatty acid metabolism, cyanoamino acid metabolism, glyceride metabolism, and plasmalogen synthesis.ConclusionIn summary, high temperature disrupted the homeostatic balance of the intestinal microbiota in mice and also induced significant alterations in intestinal metabolites. This study provides a basis for treating intestinal disorders caused by elevated temperature in humans and animals and can further formulate nutritional countermeasures to reduce heat stress-induced damage.

Supplemental Zinc exerts a positive effect against the heat stress damage in intestinal epithelial cells: Assays in a Caco-2 model

• The effect of Zn in Caco-2 cells under heat stress was investigated for first time. • Zn pretreatment induces beneficial effects on cell viability and membrane integrity. • ROS generation and antioxidant enzyme defense indicate antioxidant power of Zn. • Supplemental Zn partly prevent heat stress-induced damage in TEER and permeability. Heat stress (HS) lead to intestinal epithelial damage through induction of cellular oxidative stress. Since Zn is an essential nutrient with antioxidant properties that is involved in maintaining the intestinal barrier function, this study aimed to examine the effects of Zn supplementation in the intestinal epithelium under HS. Caco-2 cells were pre-incubated with Zn (50 and 100 µM) prior to control (37 °C) or HS conditions (41–42 °C) for 24 h. Zn pretreatment reduced the lactate dehydrogenase release, a parameter reflecting the cell membrane damage. Zn supplementation alleviated the ROS generation in cells exposed to HS and promoted the activity of antioxidant enzymes. Moreover, Zn (specially 50 µM) partly attenuated the HS induced damage on monolayer integrity as measured by transepithelial electrical resistance and phenol red permeability. Therefore, Zn supplementation had beneficial effects on the intestinal damage evoked by high temperatures and may be a useful nutritional strategy against global warming.

Responses and coping methods of different testicular cell types to heat stress: overview and perspectives.

To facilitate temperature adjustments, the testicles are located outside the body cavity. In most mammals, the temperature of the testes is lower than the body temperature to ensure the normal progression of spermatogenesis. Rising temperatures affect spermatogenesis and eventually lead to a decline in male fertility or even infertility. However, the testes are composed of different cell types, including spermatogonial stem cells (SSCs), spermatocytes, spermatozoa, Leydig cells, and Sertoli cells, which have different cellular responses to heat stress. Recent studies have shown that using different drugs can relieve heat stress-induced reproductive damage by regulating different signaling pathways. Here, we review the mechanisms by which heat stress damages different cells in testes and possible treatments.

Hydrolyzed camel whey protein alleviated heat stress-induced hepatocyte damage by activated Nrf2/HO-1 signaling pathway and inhibited NF-κB/NLRP3 axis.

Liver damage is the most severe complication of heat stress (HS). Hydrolyzed camel whey protein (CWP) possesses bioactive peptides with obviously antioxidant and anti-inflammatory activities. The current study aims to investigate whether CWP that is hydrolyzed by a simulated gastrointestinal digestion process, named S-CWP, protects BRL-3A hepatocytes from HS-induced damage via antioxidant and anti-inflammatory mechanisms. BRL-3A cells were pretreated with S-CWP before being treated at 43°C for 1h, and the levels of the cellular oxidative stress, inflammation, apoptosis, biomarkers for liver function, the activities of several antioxidant enzymes, and the cell viability were analyzed. The expression level of pivotal proteins in correlative signaling pathways was evaluated by western blotting. We confirmed that S-CWP alleviated HS-induced hepatocytes oxidative stress by decreased reactive oxygen species (ROS), nitric oxide (NO), 8-Hydroxy-2'-deoxyguanosine (8-OHdG), lipid peroxidation (LPO), protein carbonylation (PCO), and the activities of NADPH oxidase while enhanced superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), heme oxygenase-1 (HO-1) activities, and GSH content. S-CWP suppressed HS-induced inflammatory response by reducing the phosphorylation of NF-κB p65, the expression of NLRP3, and caspase-1 and finally alleviated caspase-3-mediated apoptosis. S-CWP also alleviated HS-induced hepatocyte injury by reducing alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels and restoring Heat Shock Protein 70 (HSP70) expression. Furthermore, S-CWP treatment significantly enhanced the expression of NF-E2-related nuclear factor erythroid-2 (Nrf2) and HO-1. The antioxidant and anti-inflammatory effects of S-CWP were weakened by ML385, a specific Nrf2 inhibitor. Additionally, zinc protoporphyrin (ZnPP), a specific HO-1 inhibitor, significantly reversed S-CWP-induced reduction in the phosphorylation of NF-κB p65. Thus, our results revealed that S-CWP protected against HS-induced hepatocytes damage via activating the Nrf2/HO-1 signaling pathway and inhibiting NF-κB/NLRP3 axis.

Synthetic strigolactone (rac-GR24) alleviates the adverse effects of heat stress on seed germination and photosystem II function in lupine seedlings

There is increasing experimental evidence that strigolactones, a class of carotenoid-derived sesquiterpenoid hormones, and their downstream signal components play a role in plant resilience to abiotic stress. Strigolactones positively influence plant coping mechanisms in response to abiotic stressors like drought and high salinity. In this study, we examined the effects of rac-GR24 (a synthetic strigolactone analog) and strigolactone inhibitors on the physiological and molecular responses associated with thermotolerance during seed germination and seedling development in Lupinus angustifolius under heat stress. Photosystem I & II functions were also evaluated via Chl a fluorescence transient analysis in heat stressed lupine seedlings. Our results suggest a putative role for GR24 in mediating tolerance to heat stress during seed germination and seedling development albeit these responses appeared independent of D14-mediated signalling. Seeds primed with GR24 had the highest of all germination indices, enhanced proline content and reduced peroxidation of lipids. GR24 also enhanced the activities of enzymes of the antioxidant and glyoxalase systems in lupine seedlings. The JIP-test indicated that GR24 conferred resistance to heat stress-induced damage to the oxygen evolution complex while also preventing the inactivation of PSII reaction centres thus ensuring PSII thermotolerance.

Melatonin Suppressed the Heat Stress-Induced Damage in Wheat Seedlings by Modulating the Antioxidant Machinery.

Melatonin (N-acetyl-5-methoxytryptamine) is a pleiotropic signaling molecule that plays a crucial role in the regulation of various environmental stresses, including heat stress (HS). In this study, a 100 μM melatonin (MT) pretreatment followed by exposure to heat stress for different time periods was found to efficiently reduce oxidative stress by preventing the over-accumulation of hydrogen peroxide (H2O2), lowering the lipid peroxidation content (malondialdehyde (MDA) content), and increasing proline (Pro) biosynthesis. Moreover, the activities of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), were increased substantially in MT-pretreated wheat seedlings. The presence of MT significantly improved the heat tolerance of wheat seedlings by modulating their antioxidant defense system, activating the ascorbate–glutathione (AsA–GSH) cycle comprising ascorbate peroxidase (APX), and increasing glutathione reductase (GR) activities. It also held the photosynthetic machinery stable by increasing the chlorophyll content. Enhancement in the endogenous MT contents was also observed in the MT+HS-treated plants. Furthermore, the expression of reactive oxygen species (ROS)-related genes TaSOD, TaPOD, and TaCAT, and anti-stress responsive genes, such as TaMYB80, TaWRKY26, and TaWRKY39, was also induced in MT-treated seedlings. Due to these notable changes, an improvement in stress resistance was observed in MT-treated seedlings compared with control. Taken together, our findings suggest that MT can play a key role in boosting the stress tolerance of plants by modulating the antioxidant defense system and regulating the transcription of stress-responsive genes.

Protective effects of dietary mannan oligosaccharide on heat stress-induced hepatic damage in broilers.

Heat stress is a major concern in broiler's production, which can damage liver of broilers. This study investigated the protective effects of mannan oligosaccharide (MOS) on heat stress-induced hepatic injury in broilers. A total of 144 day-old male chicks were allocated into three treatment groups. Broilers raised under normal ambient temperature were fed a basal diet (control group), and broilers under heat stress (32-33°C for 8h daily) were given the basal diet supplemented without MOS (heat stress group) or with 1g/kg MOS (MOS group) for 42days. Compared with the control group, heat stress reduced liver weight, whereas increased aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the serum. It also reduced glutathione peroxidase (GSH-Px) activity in the serum and liver, GSH content, and superoxide dismutase (SOD) activity in the liver, but increased malondialdehyde (MDA) concentration in the serum and liver. Dietary MOS decreased serum ALT activity in heat-stressed broilers. MOS inclusion also decreased serum MDA content, but elevated hepatic GSH-Px and SOD activities, with MDA content and GSH-Px activity still being different from the control group, and SOD activity being similar to the control group. Heat stress increased concentrations of tumor necrosis factor α (TNF-α) in the serum and liver, interleukin-1β (IL-1β) in the liver, and mRNA abundances of HSP70, TLR4, MyD88, TNF-α, and IL-1β in the liver of broilers. Serum TNF-α content and mRNA abundances of hepatic TLR4 and TNF-α in MOS group were lower than the heat stress group, whereas these indexes were still higher than the control group. Our results indicated that dietary MOS ameliorated hepatic damage in heat-stressed broilers through alleviation of oxidative stress and inflammation.