GO-PEG Represses the Progression of Liver Inflammation via Regulating the M1/M2 Polarization of Kupffer Cells.

As a highly promising nanomaterial, exploring the impact of the liver, a vital organ, stands out as a crucial focus in the examination of its biological effects. Kupffer cells (KCs) are one of the first immune cells to contact with exotic-substances in liver. Therefore, this study investigates the immunomodulatory effects and mechanisms of polyethylene glycol-modified graphene oxide (GO-PEG) on KCs. Initial RNA-seq and KEGG pathway analyses reveal the inhibition of the TOLL-like receptor, TNF-α and NOD-like receptor pathways in continually stimulated KCs exposed to GO-PEG. Subsequent biological experiments validate that a 48-hour exposure to GO-PEG alleviates LPS-induced KCs immune activation, characterized by a shift in polarization from M1 to M2. The underlying mechanism involves the absorption of double-stranded RNA/single-stranded RNA, inhibiting the activation of TLR3 and TLR7 in KCs. Employing a Kupffer/AML12 cell co-culture model and animal studies, it is observed that GO-PEG indirectly inhibit oxidative stress, mitochondrial dysfunction, and apoptosis in AML12 cells, partially mitigating systemic inflammation and preserving liver tissue/function. This effect is attributed to the paracrine interaction between KCs and hepatocytes. These findings suggest a meaningful and effective strategy for treating liver inflammation, particularly when combined with anti-inflammatory drugs.

Research advance of occupational exposure risks and toxic effects of semiconductor nanomaterials.

In recent years, semiconductor nanomaterials, as one of the most promising and applied classes of engineered nanomaterials, have been widely used in industries such as photovoltaics, electronic devices, and biomedicine. However, occupational exposure is unavoidable during the production, use, and disposal stages of products containing these materials, thus posing potential health risks to workers. The intricacies of the work environment present challenges in obtaining comprehensive data on such exposure. Consequently, there remains a significant gap in understanding the exposure risks and toxic effects associated with semiconductor nanomaterials. This paper provides an overview of the current classification and applications of typical semiconductor nanomaterials. It also delves into the existing state of occupational exposure, methodologies for exposure assessment, and prevailing occupational exposure limits. Furthermore, relevant epidemiological studies are examined. Subsequently, the review scrutinizes the toxicity of semiconductor nanomaterials concerning target organ toxicity, toxicity mechanisms, and influencing factors. The aim of this review is to lay the groundwork for enhancing the assessment of occupational exposure to semiconductor nanomaterials, optimizing occupational exposure limits, and promoting environmentally sustainable development practices in this domain.

Safety assessment of graphene oxide and microcystin-LR complex: a toxicological scenario beyond physical mixture.

BACKGROUND: Nanomaterials have been widely used in electrochemistry, sensors, medicine among others applications, causing its inevitable environmental exposure. A raising question is the "carrier" effect due to unique surface properties of nanomaterials, which may collectively impact the bioavailability, toxicokinetic, distribution and biological effects of classic toxicants. Noteworthy, this aspect of information remains largely unexplored. METHODS: Here, we deliberately selected two entities to mimic this scenario. One is graphene oxide (GO), which is made in ton quantity with huge surface-area that provides hydrophilicity and π-π interaction to certain chemicals of unique structures. The other is Microcystin-LR (MCLR), a representative double-bond rich liver-toxic endotoxin widely distributed in aquatic-system. Firstly, the adsorption of GO and MCLR after meeting under environmental conditions was explored, and then we focused on the toxicological effect and related mechanism of GO-MCLR complex on human skin cutin forming cells (HaCaT cells) and normal liver cells (L02 cells). RESULTS: Abiotically, our study demonstrated that GO could effectively adsorb MCLR through hydrogen bonding and π-π interaction, the oxidation degree of GO-MCLR decreased significantly and surface defect level raised. Compared to GO or MCLR, GO-MCLR was found to induce more remarkable apoptosis and ferroptosis in both HaCaT and L02 cells. The underlying mechanism was that GO-MCLR induced stronger intracellular reactive oxygen species (ROS) and mtROS generation, followed by Fe2+ accumulation, mitochondrial dysfunction and cytoskeletal damage. CONCLUSIONS: These results suggest that the GO-MCLR complex formed by GO adsorption of MCLR may exhibit more toxic effects than the single material, which demonstrates the necessity for assessing nano-toxicant complexity. Our discovery may serve as a new toxicological paradigm in which nanomaterial mediated surface adsorption effects could impact the degree of cytotoxicity and toxicological mechanisms of classic toxins.

LIGHT-INDUCED RICE1 Regulates Light-Dependent Attachment of LEAF-TYPE FERREDOXIN-NADP+ OXIDOREDUCTASE to the Thylakoid Membrane in Rice and Arabidopsis.

LIR1 (LIGHT-INDUCED RICE1) encodes a 13-kD, chloroplast-targeted protein containing two nearly identical motifs of unknown function. LIR1 is present in the genomes of vascular plants, mosses, liverworts, and algae, but not in cyanobacteria. Using coimmunoprecipitation assays, pull-down assays, and yeast two-hybrid analyses, we showed that LIR1 interacts with LEAF-TYPE FERREDOXIN-NADP(+) OXIDOREDUCTASE (LFNR), an essential chloroplast enzyme functioning in the last step of photosynthetic linear electron transfer. LIR1 and LFNR formed high molecular weight thylakoid protein complexes with the TIC62 and TROL proteins, previously shown to anchor LFNR to the membrane. We further showed that LIR1 increases the affinity of LFNRs for TIC62 and that the rapid light-triggered degradation of the LIR1 coincides with the release of the LFNR from the thylakoid membrane. Loss of LIR1 resulted in a marked decrease in the accumulation of LFNR-containing thylakoid protein complexes without a concomitant decrease in total LFNR content. In rice (Oryza sativa), photosynthetic capacity of lir1 plants was slightly impaired, whereas no such effect was observed in Arabidopsis thaliana knockout mutants. The consequences of LIR1 deficiency in different species are discussed.

OsHAC4 is critical for arsenate tolerance and regulates arsenic accumulation in rice.

Soil contamination with arsenic (As) can cause phytotoxicity and elevated As accumulation in rice grain. Here, we used a forward genetics approach to investigate the mechanism of arsenate (As(V)) tolerance and accumulation in rice. A rice mutant hypersensitive to As(V), but not to As(III), was isolated. Genomic resequencing and complementation tests were used to identify the causal gene. The function of the gene, its expression pattern and subcellular localization were characterized. OsHAC4 is the causal gene for the As(V)-hypersensitive phenotype. The gene encodes a rhodanase-like protein that shows As(V) reductase activity when expressed in Escherichia coli. OsHAC4 was highly expressed in roots and was induced by As(V). In OsHAC4pro-GUS transgenic plants, the gene was expressed exclusively in the root epidermis and exodermis. OsHAC4-eGFP was localized in the cytoplasm and the nucleus. Mutation in OsHAC4 resulted in decreased As(V) reduction in roots, decreased As(III) efflux to the external medium and markedly increased As accumulation in rice shoots. Overexpression of OsHAC4 increased As(V) tolerance and decreased As accumulation in rice plants. OsHAC4 is an As(V) reductase that is critical for As(V) detoxification and for the control of As accumulation in rice. As(V) reduction, followed by As(III) efflux, is an important mechanism of As(V) detoxification.

OsCLT1, a CRT-like transporter 1, is required for glutathione homeostasis and arsenic tolerance in rice.

Arsenic (As) contamination in a paddy environment can cause phytotoxicity and elevated As accumulation in rice (Oryza sativa). The mechanism of As detoxification in rice is still poorly understood. We isolated an arsenate (As(V))-sensitive mutant of rice. Genomic resequencing and complementation identified OsCLT1, encoding a CRT-like transporter, as the causal gene for the mutant phenotype. OsCLT1 is localized to the envelope membrane of plastids. The glutathione and γ-glutamylcysteine contents in roots of Osclt1 and RNA interference lines were decreased markedly compared with the wild-type (WT). The concentrations of phytochelatin PC2 in Osclt1 roots were only 32% and 12% of that in WT after As(V) and As(III) treatments, respectively. OsCLT1 mutation resulted in lower As accumulation in roots but higher As accumulation in shoots when exposed to As(V). Under As(III) treatment, Osclt1 accumulated a lower As concentration in roots but similar As concentration in shoots to WT. Further analysis showed that the reduction of As(V) to As(III) was decreased in Osclt1. Osclt1 was also hypersensitive to cadmium (Cd). These results indicate that OsCLT1 plays an important role in glutathione homeostasis, probably by mediating the export of γ-glutamylcysteine and glutathione from plastids to the cytoplasm, which in turn affects As and Cd detoxification in rice.

Integrative Comparison of the Role of the PHOSPHATE RESPONSE1 Subfamily in Phosphate Signaling and Homeostasis in Rice.

Phosphorus (P), an essential macronutrient for all living cells, is indispensable for agricultural production. Although Arabidopsis (Arabidopsis thaliana) PHOSPHATE RESPONSE1 (PHR1) and its orthologs in other species have been shown to function in transcriptional regulation of phosphate (Pi) signaling and Pi homeostasis, an integrative comparison of PHR1-related proteins in rice (Oryza sativa) has not previously been reported. Here, we identified functional redundancy among three PHR1 orthologs in rice (OsPHR1, OsPHR2, and OsPHR3) using phylogenetic and mutation analysis. OsPHR3 in conjunction with OsPHR1 and OsPHR2 function in transcriptional activation of most Pi starvation-induced genes. Loss-of-function mutations in any one of these transcription factors (TFs) impaired root hair growth (primarily root hair elongation). However, these three TFs showed differences in DNA binding affinities and messenger RNA expression patterns in different tissues and growth stages, and transcriptomic analysis revealed differential effects on Pi starvation-induced gene expression of single mutants of the three TFs, indicating some degree of functional diversification. Overexpression of genes encoding any of these TFs resulted in partial constitutive activation of Pi starvation response and led to Pi accumulation in the shoot. Furthermore, unlike OsPHR2-overexpressing lines, which exhibited growth retardation under normal or Pi-deficient conditions, OsPHR3-overexpressing plants exhibited significant tolerance to low-Pi stress but normal growth rates under normal Pi conditions, suggesting that OsPHR3 would be useful for molecular breeding to improve Pi uptake/use efficiency under Pi-deficient conditions. We propose that OsPHR1, OsPHR2, and OsPHR3 form a network and play diverse roles in regulating Pi signaling and homeostasis in rice.

Characterization of Cell-Death-Inducing Members of the Pectate Lyase Gene Family in Phytophthora capsici and Their Contributions to Infection of Pepper.

Pectate lyases (PL) play a critical role in pectin degradation. PL have been extensively studied in major bacterial and fungal pathogens of a wide range of plant species. However, the contribution of PL to infection by oomycete pathogens remains largely unknown. Here, we cloned 22 full-length pectate lyase (PcPL) genes from a highly aggressive strain of Phytophthora capsici SD33. Of these, PVX agroinfiltration revealed that 12 PcPL genes were found to be highly induced during infection of pepper by SD33 but the induction level was twofold less in a mildly aggressive strain, YN07. The four genes with the highest transcript levels as measured by by quantitative reverse-transcription polymerase chain reaction (PcPL1, PcPL15, PcPL16, and PcPL20) also produced a severe cell death response following transient expression in pepper leaves but the other eight PcPL genes did not. Overexpression of these four genes increased the virulence of SD33 on pepper slightly, and increased it more substantially during infection of tobacco. Overexpression of the genes in YN07 restored its aggressiveness to near that of SD33. Gene silencing experiments with the 12 PcPL genes produced diverse patterns of silencing of PcPL genes, from which it could be inferred from regression analysis that PcPL1, PcPL16, and PcPL20 could account for nearly all of the contributions of the PcPL genes to virulence.

Pulmonary hazard identifications of Graphene family nanomaterials: Adverse outcome pathways framework based on toxicity mechanisms.

Graphene-family nanomaterials (GFNs) are revolutionary new nanomaterials that have attracted significant attention in the field of nanomaterials, but the ensuing problems lie in the potential threats to public health and the ecosystem caused by these nanomaterials. From the perspective of GFN-related health risk assessments, this study reviews the current status of GFN-induced pathological lung events with a focus on the damage caused to different biological moieties (molecular, cellular, tissue, and organ) and the mechanistic relationships between different toxic endpoints. These multiple sites of damage were matched with existing adverse outcome pathways (AOPs) in an online knowledge base to obtain available molecular initiation events (MIEs), key events (KEs), and adverse outcomes (AOs). Among them, the MIEs were discussed in combination with the structure-activity relationship due to the correlation between toxicity and physical and chemical properties of GFNs. Based on the collection of information regarding MIEs, Kes, and AOs in addition to upstream and downstream causal extrapolation, the AOP framework for GFN-induced pulmonary toxicity was developed, highlighting the possible mechanisms of GFN-induced lung damage. This review intended to combine AOP with classic toxicological methods with a view to rapidly and accurately establishing a nanotoxicology infrastructure so as to contribute to public health risk assessment strategies through iteration from and animal models up to the population level.

MPA-capped CdTequantum dots induces endoplasmic reticulum stress-mediated autophagy and apoptosis through generation of reactive oxygen species in human liver normal cell and liver tumor cell.

The rapid developments in nanotechnology have brought increased attention to the safety of Quantum Dots (QDs). Exploring their mechanisms of toxicity and characterizing their toxic effects in different cell lines will help us better understand and apply QDs appropriately. This study aims to elucidate the importance of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress-induced autophagy for CdTe QDs toxicity, that is, the importance of the nanoparticles in mediating cellular uptake and consequent intracellular stress effects inside the cell. The results of the study showed that cancer cells and normal cells have different cell outcomes as a result of intracellular stress effects. In normal human liver cells (L02), CdTe QDs leads to ROS generation and prolong ER stress. The subsequent autophagosome accumulation eventually triggers apoptosis by activating proapoptotic signaling pathways and the expression of proapoptotic Bax. In contrast, in human liver cancer cells (HepG2 cells), expression of UPR restrains proapoptotic signaling and downregulates Bax, and activated protective cellular autophagy, as a result of protecting these liver cancer cells from CdTe QDs-induced apoptosis. In summary, we assess the safety of CdTe QDs and recounted the molecular mechanism underlying its nanotoxicity in normal and cancerous cells. Notwithstanding, additional detailed studies on the deleterious effects of these nanoparticles in the organisms of interest are required to ensure low-risk application.

How UV radiation and pH alternation impact graphene oxide mediated environmental toxicant adsorption and resulting safety characteristics - A toxicology study beyond a classic carrier effect.

Once released into water, the widely used graphene oxide (GO) is likely to adsorb classical environmental pollutants, exemplified by Microcystin-LR (MCLR) that is a representative double-bond rich liver-toxic endotoxin. While GO-mediated carrier effect is fairly predictable, the involvement of environmental factors like UV and pH may add additional level of sophistication as these factors may impact the adsorption capacity of GO to MCLR. Here, we firstly investigated the changes of GO structure under different UV-radiation durations and pH conditions with a view to establish the correlation in terms of MCLR adsorption onto GO. We demonstrated that GO reduction especially oxygen-containing groups reduction induced by UV- radiation caused the compromised adsorption MCLR capacity on GO. Besides, the higher pH decreased the non-biological MCLR adsorption to GO by reducing GO defect sites and increasing electrostatic repulsion. These abiotic discoveries were further investigated to compare the safety features of GO-MCLR complex. Under dark condition (pH = 7), we revealed the cytotoxicity of GO-MCLR to normal liver cells, which involved the ROS generation and cell ferroptosis caused by Fe2+ accumulation. Introduction of UV and pH alternation in environment impacted GO-mediated environmental toxicant adsorption and resulting safety characteristics, which reminded us environmental factors should not be ignored in the GO-mediated carrier effect.

Protective Effect of Ganoderma atrum Polysaccharide on Acrolein-Induced Apoptosis and Autophagic Flux in IEC-6 Cells.

This study was designed to explore the beneficial effect and mechanism of Ganoderma atrum (G. atrum) polysaccharide (PSG-1) on acrolein-induced IEC-6 cells. Our results indicated that PSG-1 significantly reduced the impairment of acrolein on cell viability, decreased oxidative stress, and enabled normal expression of tight junction (TJ) proteins that were inhibited by acrolein in IEC-6 cells. Furthermore, PSG-1 attenuated the elevation of microtubule-associated proteins light chain 3 (LC3) and Beclin 1-like protein 1 (Beclin 1) and increased the protein levels of phospho-mTOR (p-mTOR) and phospho-akt (p-akt), indicating that PSG-1 activated the mammalian target of rapamycin (mTOR) signaling pathway and alleviated acrolein-induced autophagy in IEC-6 cells. Moreover, PSG-1 markedly attenuated the acrolein-induced apoptosis, as evidenced by the increase in mitochondrial membrane potential (MMP) and B-cell lymphoma 2 (Bcl-2) expression, and the decrease in cysteine aspartate lyase (caspase)-3 and caspase-9. In addition, autophagy the inhibitor inhibited acrolein-induced TJ and apoptosis of IEC-6 cells, while the apoptosis inhibitor also inhibited acrolein-induced TJ and autophagy, suggesting that autophagy and apoptosis were mutually regulated. Taken together, the present study proved that PSG-1 could protect IEC-6 cells from acrolein-induced oxidative stress and could repair TJ by inhibiting apoptosis and autophagic flux, where autophagy and apoptosis were mutually regulated.

The Role of Acrolein in Neurodegenerative Diseases and Its Protective Strategy.

Neurodegenerative diseases are characterized by a massive loss of specific neurons, which can be fatal. Acrolein, an omnipresent environmental pollutant, is classified as a priority control contaminant by the EPA. Evidence suggests that acrolein is a highly active unsaturated aldehyde related to many nervous system diseases. Therefore, numerous studies have been conducted to identify the function of acrolein in neurodegenerative diseases, such as ischemic stroke, AD, PD, and MS, and its exact regulatory mechanism. Acrolein is involved in neurodegenerative diseases mainly by elevating oxidative stress, polyamine metabolism, neuronal damage, and plasma ACR-PC levels, and decreasing urinary 3-HPMA and plasma GSH levels. At present, the protective mechanism of acrolein mainly focused on the use of antioxidant compounds. This review aimed to clarify the role of acrolein in the pathogenesis of four neurodegenerative diseases (ischemic stroke, AD, PD and MS), as well as protection strategies, and to propose future trends in the inhibition of acrolein toxicity through optimization of food thermal processing and exploration of natural products.

"Dialogue" between Caco-2 and DCs regulated by Ganoderma atrum polysaccharide in intestinal-like Caco-2/DCs co-culture model.

The previous research has indicated that Ganoderma atrum polysaccharide (PSG-1) indirectly affects the immune function of dendritic cells (DCs) in intestinal-like Caco-2/DCs co-culture model, in which NF-κB and MAPK pathway play an essential role. To explore the interaction of Caco-2 in the interaction between the intestinal epithelium and its internal immune cells, the intestinal-like Caco-2/DCs co-culture model was developed. All transcripts of Caco-2 treated with or without PSG-1 were globally screened by RNA-seq. The expression of 452 genes regulated by PSG-1 was statistically significant, the counts of up-regulated and down-regulated genes were 198 and 256, respectively. According to KEGG analysis, tumor necrosis factor (TNF)-α and NF-κB signaling pathways of Caco-2 were selected to elucidate the mechanism of interaction between Caco-2/DCs induced by PSG-1. After the addition of TNF-α inhibitor Apremilast and NF-κB inhibitor BAY11-70821 in Caco-2, expression of cytokines (TNF-α, IL-6, IL-1ß, IL-10), chemokines (RANTES, MIP-1α, MCP-1), and the key proteins of MAPK and NF-κB pathways of DCs were all reduced. In summary, "dialogue" between Caco-2 and DCs was regulated by PSG-1 through TNF-α and NF-κB signaling pathways of Caco-2 in the model.

Characterization of the direct interaction between apple condensed tannins and cholesterol in vitro.

To provide the scientific evidences for a possible new hypocholesterolemic mechanism of apple condensed tannins (ACT), the direct interaction of ACT with cholesterol (CH) was investigated in the present study. Our results suggested that the quenching of ACT fluorescence by CH was carried out according to a static mechanism, while the interaction between ACT and CH in vitro was a spontaneous process. ACT were capable of binding with CH directly, and the CH-binding capacity (35.9-43.9%) of ACT remarkably enhanced with the increase of ACT concentration (0.5-2.0 mg proanthocyanidin B2 equivalent/mL). Besides, spectroscopic methods and morphological analysis were used to characterize the ACT-CH coprecipitates, the findings indicated that ACT were able to precipitate CH via ionic interactions, hydrophobic interactions and intermolecular hydrogen bonds rather than covalent bonds. In conclusion, the direct interaction of ACT with CH might play a role in their CH-lowering effects in humans and animals.

Preparation, characterization and in vitro hypoglycemic activity of banana condensed tannin-inulin conjugate.

To enhance the hypoglycemic effects of inulin, banana condensed tannins (BCT) were grafted onto inulin via a free radical method to synthesize the novel BCT grafted inulin (BCT-g-inulin) complex. Spectroscopic methods, XRD, TGA, 1H NMR, GPC and morphology analyses were utilized to characterize the structural properties of the BCT-g-inulin complex, and our results confirmed the conjugation of BCT and inulin. The conjugation possibly occurred between the hydroxyl group attached at the C6 position of inulin and the C6/C8 position of flavon-3-ol units of BCT. The grafting ratio and grafting efficiency of the BCT-g-inulin complex were 357.54 ± 2.98 g kg-1 complex and 74.57 ± 1.44%, respectively. The data of the antioxidant assays indicated that the BCT-g-inulin complex showed a significantly higher antioxidant activity than native inulin. Also, the grafting reaction remarkably improved the in vitro anti-diabetic activity of inulin. The glucose adsorption capacity and glucose dialysis retardation index of the BCT-g-inulin complex were remarkably higher than those of inulin, while the BCT-g-inulin complex showed much stronger inhibitory effects against α-amylase and α-glucosidase compared with inulin. Notably, the inhibition of both α-amylase and α-glucosidase by the BCT-g-inulin complex occurred through mixed-competitive mode. On the basis of fluorescence spectroscopy, the fluorescence of α-amylase and α-glucosidase could be quenched by the BCT-g-inulin complex through a static quenching mechanism. Hence, the BCT-g-inulin complex might have the potential to be developed as an effective anti-diabetic agent.

A Ganoderma atrum polysaccharide alleviated DSS-induced ulcerative colitis by protecting the apoptosis/autophagy-regulated physical barrier and the DC-related immune barrier.

Polysaccharides are one of the main active substances in Ganoderma atrum (G. atrum). The purpose of this study was to explore the protective effect of a G. atrum polysaccharide (PSG-1) on DSS-induced colitis and the underlying mechanism. The results showed that PSG-1 could maintain the integrity of the intestinal structure by promoting the expression of goblet cells and levels of tight junction proteins in the colon of DSS-induced colitis mice. Furthermore, PSG-1 relieved the inhibition of Bcl-2 and the overexpression of caspase-3 and caspase-9 caused by DSS. Simultaneously, PSG-1 restored the expression of Atg5, Atg7 and beclin-1 and inhibited the p-akt and p-mTOR levels, suggesting that PSG-1 promoted autophagy via the Akt/mTOR pathway. Moreover, PSG-1 inhibited the content of DCs in the colon and modulated the expression of IL-10 in DCs. In conclusion, PSG-1 alleviated DSS-induced ulcerative colitis by protecting the apoptosis/autophagy-regulated physical barrier and the DC-related immune barrier.

Regulatory effects of Ganoderma atrum polysaccharides on LPS-induced inflammatory macrophages model and intestinal-like Caco-2/macrophages co-culture inflammation model.

Lipopolysaccharide (LPS)-induced inflammatory macrophages model and intestinal-like Caco-2/macrophages co-culture inflammation model were established to evaluate the anti-inflammatory effect and underlying mechanism of Ganoderma atrum polysaccharides (PSG-1). It was found that PSG-1 reduced LPS-induced secretion of pro-inflammatory cytokine (TNF-α, IL-6 and IL-1ß), ROS levels, and inhibited the expression of COX-2 in LPS-stimulated inflammatory macrophages model and intestinal-like Caco-2/macrophages co-culture inflammation model. Furthermore, PSG-1 suppressed the LPS-induced activation of MAPKs signaling pathways, and regulated oxidative stress by activating the Nrf2/Keap1 signaling pathways. These above results indicated that PSG-1 not only has a direct anti-inflammatory effect in LPS-induced inflammatory macrophages model, but also has an indirect anti-inflammatory effect in intestinal-like Caco-2/macrophages co-culture inflammation model. These findings provide new insight of the mechanism underlying the anti-inflammatory activities of PSG-1, and facilitated the expansion of the application of PSG-1 in natural functional food.

Protective effect of Ganoderma atrum polysaccharide on acrolein-induced macrophage injury via autophagy-dependent apoptosis pathway.

The study aimed to investigate the protective effect and underlying mechanism of Ganoderma atrum (G. atrum) polysaccharide (PSG-1) on macrophage injury induced by acrolein. The results showed that PSG-1 restored the cell viability damaged by acrolein. In addition, PSG-1 significantly reduced the acrolein-induced occurrence of apoptosis via increase of Bcl-2 expression, mitochondrial membrane potential (MMP), decrease of ROS, cytochrome c (Cyt-C), caspase-3, caspase-9. Moreover, the overexpressions of autophagy-related proteins (LC3, Beclin-1, Atg7 and Atg5) were suppressed by PSG-1, which demonstrated that PSG-1 inhibited autophagy in acrolein treated macrophage. Beside, PSG-1 significantly elevated the expression level of p-mTOR, suggested that PSG-1 mediated autophagy through mTOR pathway. Furthermore, inhibitor of autophagy could inhibit apoptosis in acrolein-induced macrophage, suggesting that autophagy may be involved in the regulation of apoptosis. In summary, the present study demonstrated that PSG-1 protected acrolein-induced macrophage injury via autophagy-dependent apoptosis.

Leptin siRNA promotes ovarian granulosa cell apoptosis and affects steroidogenesis by increasing NPY2 receptor expression.

Leptin has been found to be involved in the ovarian granulosa cell apoptosis and steroidogenesis. Loss of neuropeptide Y (NPY) can correct the obesity syndrome of mutant mice lacking of leptin (ob/ob). However, the association of NPY and leptin in ovarian granulosa cells and ovarian steroidogenesis has not been investigated. Here, C57BL/6J ob/ob mice and C57BL/6J (control) mice were intraperitoneally injected with PBS, leptin (0.4µg/g bodyweight) or BIIE0246 (NPY2 receptor [NPY2R] antagonist, 30µg/kg bodyweight) every day for 15days. We found that NPY2R mRNA expression in mouse ovary was suppressed by leptin treatment, but increased by leptin deficiency. Leptin or BIIE0246 treatment significantly increased E2, but notably decreased progesterone in both mice. A lower level of E2 and a higher level of progesterone was observed in ob/ob mice than in control mice. Further, we then knocked down leptin expression in human ovarian granulosa cells by siRNA transfection and treated the cells with DMSO or BIIE0246. In vitro experiments confirmed the findings in mice. siLeptin treatment decreased the secretion of E2, anti-Mullerian hormone (AMH), insulin-like growth factor (IGF)-1 and transforming growth factor (TGF)-ß, and the cell proliferation, but increased the secretion of progesterone and cell apoptosis. Western blotting analysis of PCNA, Bcl-2 and Bax confirmed the results of cell proliferation and apoptosis. Activation of JAK2 and STAT3 was also suppressed by knocking down leptin. All the effects of siLeptin on ovarian granulosa cells were partially reversed by BIIE0246. In conclusion, knockdown of leptin significantly affected ovarian steroidogenesis and ovarian function through NPY. siLeptin transfection impaired the activation of JAK2/STAT3 and contributed to ovarian granulosa cell apoptosis partially through up-regulating NPY2R expression.