Changes in the chemical properties and metabolite profiling of fish sauce prepared from underutilized large yellow croaker roes during fermentation at different temperatures.

Fish sauce is a popular aquatic condiment with unique flavor. In this study, the changes in the chemical properties and metabolite profiling of fish sauce from large yellow croaker roes during fermentation at different temperatures were revealed. The results found that the contents of total acid, amino acid nitrogen, total soluble nitrogen and soluble salt-free solids of fish sauce fermented at 40 °C were higher than those in other temperatures groups (25 °C and 32 °C), while the contents of total volatile basic nitrogen were lower than other temperatures. Therefore, 40 °C was the ideal fermentation temperature for fish sauce. The metabolomics analysis showed that organic acids, amino acids, nucleotide, and lipid compounds were found to participate in the biosynthesis pathway. Compared to 25 °C and 32 °C, fermented at 40 °C could increase the abundance of metabolic substances in the fish sauce, such as sugar alcohols, L-Citrulline, L-Aspartic acid, L-Cysteine, Glutathione, and L-Arginine. These results provide a theoretical basis for the production of high-quality fish sauce and the high-value utilization of fish roes.

Insight into the effect of large yellow croaker roe phospholipids on the physical properties of surimi gel and their interaction mechanism with myofibrillar protein.

BACKGROUND: The present study aimed to investigate the effects of large yellow croaker roe phospholipids (LYCRPLs) on the physical properties of surimi gels and to clarify their interaction mechanism with myofibrillar proteins (MPs) in terms of chemical forces and the spatial conformation. RESULTS: LYCRPLs could improve the gel strength, textural properties, rheological properties and water-holding capacity of surimi gels. Moreover, the interaction mechanism between LYCRPLs with MPs was revealed through intermolecular forces, Fourier transform infrared spectroscopy and ultraviolet visible absorption spectroscopy. The findings demonstrated that LYCRPLs enhanced the surface hydrophobicity and particle size of MPs, facilitating expansion and cross-linking of MPs. CONCLUSION: These results provide a theoretical basis for improving the characteristics of surimi gels and thus facilitate the application of LYCRPLs in the aquatic food industry. © 2023 Society of Chemical Industry.

CircHERC1 promotes non-small cell lung cancer cell progression by sequestering FOXO1 in the cytoplasm and regulating the miR-142-3p-HMGB1 axis.

BACKGROUND: Noncoding RNAs such as circular RNAs (circRNAs) are abundant in the human body and influence the occurrence and development of various diseases. Non-small cell lung cancer (NSCLC) is one of the most common malignant cancers. Information on the functions and mechanism of circRNAs in lung cancer is limited; thus, the topic needs more exploration. The purpose of this study was to identify aberrantly expressed circRNAs in lung cancer, unravel their roles in NSCLC progression, and provide new targets for lung cancer diagnosis and therapy. METHODS: High-throughput sequencing was used to analyze differential circRNA expression in patients with lung cancer. qRT‒PCR was used to determine the level of circHERC1 in lung cancer tissues and plasma samples. Gain- and loss-of-function experiments were implemented to observe the impacts of circHERC1 on the growth, invasion, and metastasis of lung cancer cells in vitro and in vivo. Mechanistically, dual luciferase reporter assays, fluorescence in situ hybridization (FISH), RNA immunoprecipitation (RIP) and RNA pull-down experiments were performed to confirm the underlying mechanisms of circHERC1. Nucleocytoplasmic localization of FOXO1 was determined by nucleocytoplasmic isolation and immunofluorescence. The interaction of circHERC1 with FOXO1 was verified by RNA pull-down, RNA immunoprecipitation (RIP) and western blot assays. The proliferation and migration of circHERC1 in vivo were verified by subcutaneous and tail vein injection in nude mice. RESULTS: CircHERC1 was significantly upregulated in lung cancer tissues and cells, ectopic expression of circHERC1 strikingly facilitated the proliferation, invasion and metastasis, and inhibited the apoptosis of lung cancer cells in vitro and in vivo. However, knockdown of circHERC1 exerted the opposite effects. CircHERC1 was mainly distributed in the cytoplasm. Further mechanistic research indicated that circHERC1 acted as a competing endogenous RNA of miR-142-3p to relieve the repressive effect of miR-142-3p on its target HMGB1, activating the MAPK/ERK and NF-κB pathways and promoting cell migration and invasion. More importantly, we found that circHERC1 could bind FOXO1 and sequester it in the cytoplasm, adjusting the feedback AKT pathway. The accumulation of FOXO1 in the cytosol and nuclear exclusion promoted cell proliferation and inhibited apoptosis. CircHERC1 is a new circRNA that promotes tumor function in NSCLC and may serve as a potential prognostic biomarker and therapeutic target for NSCLC. CONCLUSIONS: CircHERC1 is a new circRNA that promotes tumor function in NSCLC and may serve as a potential diagnosis biomarker and therapeutic target for NSCLC. Our findings indicate that circHERC1 facilitates the invasion and metastasis of NSCLC cells by regulating the miR-142-3p/HMGB1 axis and activating the MAPK/ERK and NF-κB pathways. In addition, circHERC1 can promote cell proliferation and inhibit apoptosis by sequestering FOXO1 in the cytoplasm to regulate AKT activity and BIM transcription.

Effect of DHA-Enriched Phospholipids from Fish Roe on Rat Fecal Metabolites: Untargeted Metabolomic Analysis.

Lipid metabolism disorder has become an important hidden danger threatening human health, and various supplements to treat lipid metabolism disorder have been studied. Our previous studies have shown that DHA-enriched phospholipids from large yellow croaker (Larimichthys Crocea) roe (LYCRPLs) have lipid-regulating effects. To better explain the effect of LYCRPLs on lipid regulation in rats, the fecal metabolites of rats were analyzed from the level of metabolomics in this study, and GC/MS metabolomics measurements were performed to figure out the effect of LYCRPLs on fecal metabolites in rats. Compared with the control (K) group, 101 metabolites were identified in the model (M) group. There were 54, 47, and 57 metabolites in the low-dose (GA), medium-dose (GB), and high-dose (GC) groups that were significantly different from that of group M, respectively. Eighteen potential biomarkers closely related to lipid metabolism were screened after intervention with different doses of LYCRPLs on rats, which were classified into several metabolic pathways in rats, including pyrimidine metabolism, the citric acid cycle (TCA cycle), the metabolism of L-cysteine, carnitine synthesis, pantothenate and CoA biosynthesis, glycolysis, and bile secretion. L-cysteine was speculated to be a useful biomarker of LYCRPLs acting on rat fecal metabolites. Our findings indicated that LYCRPLs may regulate lipid metabolism disorders in SD rats by activating these metabolic pathways.

Development of a Golgi-targeted superoxide anion fluorescent probe for elucidating protein GOLPH3 function in myocardial ischemia-reperfusion injury.

Superoxide anion (O2•-) is an important reactive oxygen species (ROS) and participates in various physiological and pathological processes in the organism. The O2•- burst induced by ischemia-reperfusion (I/R) is associated with cardiovascular disease and promotes the cell apoptosis. In this work, a turn-on type Golgi-targeting fluorescent probe Gol-Cou-O2•- was rationally designed for sensitive and selective detection of O2•-. The minimum detection limit concentration for O2•- was about 3.9 × 10-7 M in aqueous solution. Gol-Cou-O2•- showed excellent capacity of detecting exogenous and endogenous O2•- in living cells and zebrafish, and was also used to capture the up-regulated O2•- level during the duration of I/R process in cardiomyocytes. Golgi Phosphoprotein 3 (GOLPH3) is a potential Golgi stress marker protein and plays a key role in cells apoptosis during I/R. The fluorescence imaging and flow cytometry assay results indicated that silencing GOLPH3 through siRNA could give rise to the down-regulated O2•- level and alleviation of apoptosis in I/R myocardial cells. Thus, development of Gol-Cou-O2•- provides a diagnostic tool for myocardial oxidative stress injury and distinct insights on roles of GOLPH3 in myocardial I/R injury.

Development of a Golgi-targeted fluorescent chemosensor for detecting ferrous ions overload under Golgi stress.

Ferrous ion (Fe2+) is a crucial metal ion in the body and participates in the diseases related to oxidation and reduction. Golgi apparatus is the main subcellular organelle of Fe2+ transport in cells, and the stability of its structure is related to the Fe2+ at an appropriate concentration. In this work, a turn-on type Golgi-targeting fluorescent chemosensor Gol-Cou-Fe2+ was rationally designed for sensitive and selective detection of Fe2+. Gol-Cou-Fe2+ showed excellent capacity of detecting exogenous and endogenous Fe2+ in HUVEC and HepG2 cells. It was used to capture the up-regulated Fe2+ level during the hypoxia. Moreover, the fluorescence of sensor was enhanced over time under Golgi stress combining with the reduce of Golgi matrix protein GM130. However, elimination of Fe2+ or addition of nitric oxide (NO) would restore the fluorescence intensity of Gol-Cou-Fe2+ and the expression of GM130 in HUVEC. Thus, development of chemosensor Gol-Cou-Fe2+ provides a new window for tracking Golgi Fe2+ and elucidating Golgi stress-related diseases.

Redox-Reversible Near-Infrared Fluorescent Probe for Imaging of Acute Kidney Oxidative Injury and Remedy.

Drug-induced acute kidney injury (DIAKI) is associated with high morbidity and mortality. It remains a diagnostic and therapeutic dilemma due to failure of providing unambiguous real-time feedback on nephrotoxicity, which is regarded as a serious problem in clinics. Herein, we report a reversible fluorescence probe, NRN, to monitor the ONOO-/GSH in an acute kidney injury model. The NRN near-infrared fluorescent probe features a big Stokes shift (83 nm), which was oxidized by ONOO- and reduced by succussive glutathione (GSH) with excellent selectivity and good sensitivity (detection limit: 418 nM and 0.28 mM, respectively). Taking the reversibility of NRN toward ONOO- and GSH, real-time evaluations in vivo with cisplatin (CP) alone and CP combined with acetaminophen-stimulated acute kidney injury and the following remedy process with l-carnitine were realized for the first time. The experiments revealed that acute kidney injury caused by combined drugs might be more serious and irreversible under certain conditions. Therefore, NRN could act as a potential tool for understanding oxidative stress-related DIAKI disease processes.

Enhancing the Selectivity of Leucine Aminopeptidase Near-Infrared Fluorescent Probes for Assisting in Surgical Tumor Resection.

Selective fluorescence imaging of analytes is a challenge for monitoring diseases as homologues interfere with the imaging agents. Leucine aminopeptidase (LAP), a kind of protease, is related to tumor pathogenesis. The known LAP fluorescent probes based on leucine recognition have limited selectivity. Herein, a selective t-butyl-alanine recognition unit for LAP through the ligand regulation strategy is prepared as a new near-infrared (NIR) fluorescent probe (DCM-LAP) having a large Stokes shift of 214 nm and a high sensitivity with a detection limit of 168 mU/L. DCM-LAP has an enhanced response toward LAP with NIR fluorescence at 656 nm based on intramolecular charge transfer. The probe is selective without being interfered with by biological enzymes including the aminopeptidase N (APN). DCM-LAP can image LAP activity in living cells. It can also visualize the cell invasion and migration processes. DCM-LAP is employed in the real-time imaging of LAP in tumor-bearing nude mice and guides in the accurate resection of breast tumors. It also distinguishes tumor tissues from normal with a high tumor-to-normal ratio (9.8). The DCM-LAP probe can thus assist in the investigations of LAP-associated clinical disease.

A mitochondrial-targeted near-infrared fluorescent probe for visualizing the fluctuation of hypochlorite acid in idiopathic pulmonary fibrosis mice.

Idiopathic pulmonary fibrosis (IPF) is a chronic inflammatory disease destroying lungs irreversibly with high mortality rates. There are challenges in diagnosing IPF and treating it at an early stage. Mounting evidence suggests that hypochlorous acid (HClO) can help in diagnosing inflammation and relevant conditions. Pulmonary fibrosis is linked to the mitochondrial oxidative stress where excessive HClO production is a key molecular mechanism. Measuring mitochondrial HClO levels assists in the investigations of how the mitochondrial oxidative stress affects IPF. Herein, NIR-PTZ-HClO was developed and optimized as a probe for detecting fluctuations in HClO concentrations of cells and mice models through near-infrared (NIR) fluorescence. The probe featured large Stokes shift of 150 nm, NIR turn-on signal at 650 nm, high sensitivity (45-fold) and quick HClO detection (2 s). The probe is selective for HClO in the presence of range of other analytes. NIR-PTZ-HClO visualized both endogenous and exogenous HClO in living cells (RAW264.7, H460 and A549). The probe monitored HClO in mice models with IPF and moreover the HClO profile could be tracked during the IPF process. The probe also detected precipitous decrease in HClO levels in IPF mice treated with OFEV. NIR-PTZ-HClO probe has thus the potential for earlier diagnosis of lung fibrosis, thereby improving the treatment efficacy.

circTAB2 inhibits lung cancer proliferation, migration and invasion by sponging miR-3142 to upregulate GLIS2.

Circular RNAs (circRNAs) are a specialized circular structure, are deregulated in cancers and play essential roles in biological processes involved in tumor progression. However, the mechanism by which circRNAs affect lung tumorigenesis and progression remains largely unexplored. To investigate the role of circRNA in lung cancer, circRNA expression profile was screened by bioinformatics analysis. The levels of circTAB2, miR-3142, and GLIS family zinc finger 2 (GLIS2) were measured by quantitate real-time (qRT-PCR) or western blot. Cell proliferation, apoptosis, migration and invasion were detected by EdU, flow cytometry, and transwell assays, respectively. Bioinformatics, western blot, RIP, pull down, dual luciferase reporter and rescue experiments were used to verify the direct relationship between miR-3142 and circTAB2 or GLIS2. The xenograft assays were used to assess the role of circTAB2 in vivo.CircTAB2 exhibited low expression in cancer tissues. Gain and loss-of-function assays indicated that circTAB2 could inhibit cell proliferation, migration and invasion. Functional studies revealed that circTAB2 acted as a miRNA sponge, directly interacted with miR-3142 and consequently regulated GLIS2 /AKT. Taken together, circTAB2 serves as an inhibitory role in lung cancer through a novel circTAB2 /miR-3142 /GLIS2 /AKT pathway and could be exploited a novel marker in lung cancer.

Antioxidative properties of fish roe peptides combined with polyphenol on the fish oil oleogel.

BACKGROUND: This study aimed to investigate the effects of large yellow croaker (Larimichthys crocea) roe protein hydrolysate (LYCPH)-polyphenol (catechin (CA), gallic acid (GA), and tannic acid (TA)) conjugates on the oxidative stability of fish oil in an oleogel system. RESULTS: Scanning electron microscopy and Fourier transform infrared spectroscopy suggested that the LYCPH-polyphenol conjugates were nearly spherical and non-covalent and that covalent effects could coexist between LYCPH and polyphenols. LYCPH-TA exhibited the highest ABTS scavenging, reducing capacities, and emulsifying stability. Raman spectra and chemometrics revealed that LYCPH-TA loaded with oleogels had the best oxidative stability. Additionally, 32 volatile compounds were identified in fish oil by headspace gas chromatography-ion mobility spectrometry. CONCLUSION: Overall, this study demonstrated that fish oil oleogels loaded with LYCPH-polyphenol conjugates could inhibit fish oil oxidation. © 2022 Society of Chemical Industry.

Integrated lipidomic and transcriptomic analyses reveal the mechanism of large yellow croaker roe phospholipids on lipid metabolism in normal-diet mice.

Large yellow croaker roe phospholipids (LYCRPLs) could regulate the accumulation of triglycerides and blood lipid levels. However, there exists little research on the mechanism of LYCRPLs on lipid metabolism in normal-diet mice. In this work, the mice on a normal diet were given low-dose, medium-dose, and high-dose LYCRPLs by intragastric administration for 6 weeks. At the same time, the physiological and biochemical indicators of the mice were determined, and the histomorphological observation of the liver and epididymal fat was carried out. In addition, we examined the gene expression and lipid metabolites in the liver of mice using transcriptomic and lipidomic and performed a correlation analysis. The results showed that LYCRPLs regulated the lipid metabolism of normal-diet mice by affecting the expression of the glycerolipid metabolism pathway, insulin resistance pathway, and cholesterol metabolism pathway. This study not only elucidated the main pathway by which LYCRPLs regulate lipid metabolism, but also laid a foundation for exploring LYCRPLs as functional food supplements.

DHA-enriched phospholipids from large yellow croaker roe regulate lipid metabolic disorders and gut microbiota imbalance in SD rats with a high-fat diet.

Large yellow croaker roe phospholipids (LYCRPLs) have great nutritional value because they are rich in docosahexaenoic acid (DHA), which is an n-3 polyunsaturated fatty acid (n-3 PUFA). In previous research, we studied the effect of LYCRPLs on the inhibition of triglyceride accumulation at the cellular level. However, its lipid regulation effect in rats on a high-fat diet and its influence on the gut microbiota has not yet been clarified. In this study, a high-fat diet was used to induce the lipid metabolism disorder in SD rats, and simvastatin, low-dose, medium-dose and high-dose LYCRPLs were given by intragastric administration for 8 weeks. The rats' body weight, food intake, organ index, blood biochemical indicators, epididymal fat tissue and liver histopathology were compared and analyzed. High-throughput 16S rRNA gene sequencing technology and bioinformatics analysis technology were also used to analyze the diversity of gut microbiota in rats. We found that LYCRPLs can significantly regulate lipid metabolism, and improve the gut microbiota disorder induced in rats by a high-fat diet. These results can lay a foundation for the study of the regulation mechanism of LYCRPLs lipid metabolism, and also provide a theoretical basis for the development of LYCRPLs as functional food additives and excipients with hypolipidemic effects.

Quality assessment of large yellow croaker (Larimichthys crocea) roe oil before and after refining.

This research aimed to assess the quality of the large yellow croaker (Larimichthys crocea) roe oil before and after refining. The crude and refined L. crocea roe oils were compared based on their peroxide value (PV), acid value (AV), iodine value (IV), saponification value (SV), and fatty acid composition. Furthermore, the volatile compounds were identified and analyzed via gas chromatography-mass spectroscopy (GC-MS) and electronic nose (E-nose) analysis. Meanwhile, the flavor fingerprint was established via headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS). The results showed that the PV, AV, IV, and SV of the refined oil were 4.44 ± 0.04 mmol kg-1, 2.86 ± 0.01 mgKOH g-1, 163.1 ± 0.8 g/100 g, and 222.9 ± 0.7 mg g-1, respectively. The docosahexaenoic acids (DHAs) content in the total polyunsaturated fatty acids (PUFAs) was increased. Moreover, 55 volatile compounds were identified in the refined oil; among these compounds, the contents of carboxylic acids, aldehydes, alcohols, ketones, and esters were reasonably increased, while the hydrocarbon and heterocyclic compound contents were decreased. The flavor fingerprints of the crude and refined L. crocea roe oils were established by HS-GC-IMS. The results demonstrated that the refining improved the quality of L. crocea roe oil.

Inhibition Effect of Triglyceride Accumulation by Large Yellow Croaker Roe DHA-PC in HepG2 Cells.

The phospholipids (PLs) of large yellow croaker (Pseudosciaena crocea, P. crocea) roe contain a high level of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), which can lower blood lipid levels. In previous research, PLs of P. crocea roe were found able to regulate the accumulation of triglycerides. However, none of these involve the function of DHA-containing phosphatidylcholine (DHA-PC), which is the main component of PLs derived from P. crocea roe. The function by which DHA-PC from P. crocea roe exerts its effects has not yet been clarified. Herein, we used purified DHA-PC and oleic acid (OA) induced HepG2 cells to establish a high-fat model, and the cell activity and intracellular lipid levels were then measured. The mRNA and protein expression of Fatty Acid Synthase (FAS), Carnitine Palmitoyl Transferase 1A (CPT1A) and Peroxisome Proliferator-Activated Receptor α (PPARα) in HepG2 cells were detected via RT-qPCR and western blot as well. It was found that DHA-PC can significantly regulate triglyceride accumulation in HepG2 cells, the effect of which was related to the activation of PPARα receptor activity, upregulation of CPT1A, and downregulation of FAS expression. These results can improve the understanding of the biofunction of hyperlipidemia mediated by DHA-PC from P. crocea roe, as well as provide a theoretical basis for the utilization of DHA-PC from P. crocea roe as a functional food additive.