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1.
Environ Microbiol Rep ; 16(2): e13256, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38575150

ABSTRACT

This study examined the effects of three Bacillus strains and one Saccharomyces cerevisiae strain on nitrogen transformation and microbial communities in pig and chicken manure compost. The findings revealed that the use of compound microbial inoculants increased the compost temperature, accelerated moisture reduction, enhanced cellulase activity, and stimulated the accumulation of NH4 +-N, NO3 --N, and total nitrogen (TN), resulting in a 9% increase in TN content. The abundance of Firmicutes decreased by 3.95% at the maturation phase, while Actinobacteria and Bacteroidetes increased by 1.64% and 1.85%, respectively. Inoculation led to an increase in amoA, nxrA and nifH gene copy numbers, while simultaneously reducing the abundance of nirK, nosZ and nirS genes. It also resulted in an increase in functional enzyme levels, specifically nif and amo, with a corresponding decrease in nor. Clostridium, Phascolarctobacterium, Eubacterium and Faecalibacterium from the class Clostridium, which have a significant correlation with nifH and nxrA genes, suggest their likely crucial role in nitrogen retention and fixation. Inoculation aided in the removal of pathogenic bacteria and antibiotic resistance genes (ARGs) like fluoroquinolones, nucleosides and nitroimidazole. This study provides effective theoretical support for the mechanism of nitrogen retention and fixation, and for improving the quality of compost.


Subject(s)
Composting , Microbiota , Animals , Swine , Manure , Livestock , Nitrogen , Soil , Bacteria/genetics , Microbiota/genetics
2.
J Hazard Mater ; 465: 133172, 2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38071777

ABSTRACT

Although antibiotic alternatives are widely used in livestock and poultry breeding industry after in-feed antibiotics ban, their intervention effects on antibiotic resistance genes (ARGs) in these food animals' feces remain poorly understood. Here effects of fructooligosaccharide (FOS) and astragalus polysaccharide (APS), as typical antibiotic alternatives in China, on ARGs in layer feces were estimated by performing metagenomic sequencings and fluorescence quantitative PCR. Fructooligosaccharide significantly reduced sum abundance of ARGs and mobile genetic elements (MGEs) by increasing Lactobacillus clones and reducing Escherichia clones which had relatively higher abundances of ARG subtypes and MGE subtypes in layer feces. However, at least parts of core ARGs and MGEs categories were not reduced by FOS, such as aminoglycosides- and tetracyclines-resistant genes, Tn916, Integrase, and so on. MGEs and microbiome, especially Escherichia genus and Lactobacillus genus, were the key factors affecting ARGs' sum abundance. MGEs had a higher correlation coefficient with ARGs' sum abundance than Escherichia genus and Lactobacillus genus. These findings firstly reveal the defects of antibiotic alternatives in controlling bacterial resistance in livestock and poultry breeding after in-feed antibiotics ban, and more strategies are needed to control pollutions and risks of core ARGs and MGEs in food animals' feces under a special environment.


Subject(s)
Anti-Bacterial Agents , Genes, Bacterial , Oligosaccharides , Animals , Anti-Bacterial Agents/pharmacology , Drug Resistance, Microbial/genetics , Feces , Polysaccharides
3.
Clin Transl Med ; 13(11): e1441, 2023 11.
Article in English | MEDLINE | ID: mdl-37997538

ABSTRACT

BACKGROUND: The study and synthesis of membrane organelles are becoming increasingly important, not only as simplified cellular models for corresponding molecular and metabolic studies but also for applications in synthetic biology of artificial cells and drug delivery vehicles. Lipid droplets (LDs) are central organelles in cellular lipid metabolism and are involved in almost all metabolic processes. Multiple studies have also demonstrated a high correlation between LDs and metabolic diseases. During these processes, LDs reveal a highly dynamic character, with their lipid fraction, protein composition and subcellular localisation constantly changing in response to metabolic demands. However, the molecular mechanisms underlying these functions have not been fully understood due to the limitations of cell biology approaches. Fortunately, developments in synthetic biology have provided a huge breakthrough for metabolism research, and methods for in vitro synthesis of LDs have been successfully established, with great advances in protein binding, lipid function, membrane dynamics and enzymatic reactions. AIMS AND METHODS: In this review, we provide a comprehensive overview of the assembly and function of endogenous LDs, from the generation of lipid molecules to how they are assembled into LDs in the endoplasmic reticulum. In particular, we highlight two major classes of synthetic LD models for fabrication techniques and their recent advances in biology and explore their roles and challenges in achieving real applications of artificial LDs in the future.


Subject(s)
Lipid Droplets , Metabolic Diseases , Humans , Lipid Droplets/chemistry , Lipid Droplets/metabolism , Lipid Metabolism , Endoplasmic Reticulum/metabolism , Lipids/analysis , Metabolic Diseases/metabolism
4.
Ecotoxicol Environ Saf ; 266: 115551, 2023 Nov 01.
Article in English | MEDLINE | ID: mdl-37832484

ABSTRACT

The increasing prevalence of antibiotic-resistant bacteria (ARB) from animal manure has raised concerns about the potential threats to public health. The bioconversion of animal manure with insect larvae, such as the black soldier fly larvae (BSFL, Hermetia illucens [L.]), is a promising technology for quickly attenuating ARB while also recycling waste. In this study, we investigated BSFL conversion systems for chicken manure. Using metagenomic analysis, we tracked ARB and evaluated the resistome dissemination risk by investigating the co-occurrence of antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and bacterial taxa in a genetic context. Our results indicated that BSFL treatment effectively mitigated the relative abundance of ARB, ARGs, and MGEs by 34.9%, 53.3%, and 37.9%, respectively, within 28 days. Notably, the transferable ARGs decreased by 30.9%, indicating that BSFL treatment could mitigate the likelihood of ARG horizontal transfer and thus reduce the risk of ARB occurrence. In addition, the significantly positive correlation links between antimicrobial concentration and relative abundance of ARB reduced by 44.4%. Moreover, using variance partition analysis (VPA), we identified other bacteria as the most important factor influencing ARB, explaining 20.6% of the ARB patterns. Further analysis suggested that antagonism of other bacteria on ARB increased by 1.4 times, while nutrient competition on both total nitrogen and crude fat increased by 2.8 times. Overall, these findings provide insight into the mechanistic understanding of ARB reduction during BSFL treatment of chicken manure and provide a strategy for rapidly mitigating ARB in animal manure.


Subject(s)
Diptera , Manure , Animals , Larva/genetics , Manure/analysis , Chickens/genetics , Angiotensin Receptor Antagonists , Angiotensin-Converting Enzyme Inhibitors , Diptera/genetics , Bacteria , Drug Resistance, Microbial , Genes, Bacterial , Anti-Bacterial Agents/pharmacology
5.
Bioresour Technol ; 387: 129698, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37598805

ABSTRACT

Composting generates odorous gases, including ammonia (NH3), hydrogen sulfide (H2S), and volatile organic compounds (VOCs). The Biological Trickling Filter (BTF) is effective for odor treatment, but it may have limitations with hydrophobic VOCs. In this study, a strain of Bacillus subtilis with ammonia-reducing ability, a strain of Bacillus cereus with desulfurization ability and a strain of Schizophyllum commune with the ability to degrade dimethyl disulfide were isolated and screened. The three strains were combined to create synthetic microbial consortia for enhancing odor treatment in the BTF. Compared to the activated sludge control, the BTF with synthetic microbial consortia removed 92.43% ammonia, 92.75% hydrogen sulfide. Furthermore, it demonstrated a significant improvement in the removal rates of p-methyl mercaptan, methyl sulfide, and dimethyl disulfide. High-throughput sequencing was conducted on the fillers of the synthetic microbial consortia-inoculated BTF to analyze the microbial community composition.


Subject(s)
Composting , Hydrogen Sulfide , Volatile Organic Compounds , Microbial Consortia , Ammonia , Odorants
6.
Cell Death Discov ; 9(1): 254, 2023 Jul 20.
Article in English | MEDLINE | ID: mdl-37474495

ABSTRACT

Lipid droplets (LDs) are cellular organelles comprising a core of neutral lipids (glycerides, sterols) encased within a single phospholipid membrane, responsible for storing surplus lipids and furnishing cellular energy. LDs engage in lipid synthesis, catabolism, and transport processes by interacting with other organelles (e.g., endoplasmic reticulum, mitochondria), and they play critical roles in regulating cellular stress and immunity. Recent research has uncovered that an elevated number of LDs is a hallmark of cancer cells, attributable to their enhanced lipid uptake and synthesis capacity, with lipids stored as LDs. Depletion of LDs in cancer cells induces apoptosis, prompting the emergence of small molecule antitumor drugs targeting LDs or key factors (e.g., FASN, SCD1) within the lipid synthesis pathway. Advancements in LD isolation and artificial synthesis have demonstrated their potential applicability in antitumor research. LDs extracted from murine adipose tissue and incubated with lipophilic antitumor drugs yield drug-coated LDs, which promote apoptosis in cancer cells. Furthermore, LDs have been employed as biological lenses to augment the resolution of subcellular structures (microfilaments, microtubules), facilitating the observation of intricate structures within thicker cells, including cancer cells. This review delineates the functional and metabolic mechanisms of LDs in cancer cells and encapsulates recent progress in LD-centered antitumor research, offering novel insights for tumor diagnosis and treatment.

7.
Biol Res ; 56(1): 38, 2023 Jul 12.
Article in English | MEDLINE | ID: mdl-37438836

ABSTRACT

Since the discovery, lipid droplets (LDs) have been recognized to be sites of cellular energy reserves, providing energy when necessary to sustain cellular life activities. Many studies have reported large numbers of LDs in eggs and early embryos from insects to mammals. The questions of how LDs are formed, what role they play, and what their significance is for embryonic development have been attracting the attention of researchers. Studies in recent years have revealed that in addition to providing energy for embryonic development, LDs in eggs and embryos also function to resist lipotoxicity, resist oxidative stress, inhibit bacterial infection, and provide lipid and membrane components for embryonic development. Removal of LDs from fertilized eggs or early embryos artificially leads to embryonic developmental arrest and defects. This paper reviews recent studies to explain the role and effect mechanisms of LDs in the embryonic development of several species and the genes involved in the regulation. The review contributes to understanding the embryonic development mechanism and provides new insight for the diagnosis and treatment of diseases related to embryonic developmental abnormalities.


Subject(s)
Embryonic Development , Lipid Droplets , Female , Pregnancy , Animals , Oxidative Stress , Mammals
8.
Cell Cycle ; 22(12): 1421-1433, 2023 06.
Article in English | MEDLINE | ID: mdl-37218663

ABSTRACT

During eukaryotic cell division, organelles are distributed between daughter cells through a dynamic process to ensure that cells can differentiate and perform their functions correctly. Uncovering the mode of lipid droplet (LD) distribution may help reveal the mechanism of membrane remodeling during cell division and lipid droplet function. Our results showed that LDs were equally distributed in both daughter cells during cytokinesis. Further experiments demonstrated that the key factor regulating the movement of LDs is the microtubule (MT)-resident protein KIF5B. Because the KIF5B structure lacks a hydrophilic region, we believe that there are proteins that mediate the interaction between LDs and KIF5B. Mass spectrometric detection of KIF5B-interacting proteins on the surface of LDs demonstrated that LDs were first wrapped by intermediate filaments forming a meshwork and then contacted with MTs to mediate lipid droplet movement during cytokinesis. Disruption of the homogeneous distribution of LDs may hinder cell proliferation and even lead to apoptosis.


Subject(s)
Lipid Metabolism , Microtubules , Mitosis , Microtubules/metabolism , Humans , HeLa Cells , Animals , Mice , Kinesins/metabolism
9.
Sci Total Environ ; 879: 163065, 2023 Jun 25.
Article in English | MEDLINE | ID: mdl-36966826

ABSTRACT

The increasing prevalence of antibiotic resistance genes (ARGs) in animal manure has attracted considerable attention because of their potential contribution to the development of multidrug resistance worldwide. Insect technology may be a promising alternative for the rapid attenuation of ARGs in manure; however, the underlying mechanism remains unclear. This study aimed to evaluate the effects of black soldier fly (BSF, Hermetia illucens [L.]) larvae conversion combined with composting on ARGs dynamics in swine manure and to uncover the mechanisms through metagenomic analysis. Compared to natural composting (i.e. without BSF), BSFL conversion combined with composting reduced the absolute abundance of ARGs by 93.2 % within 28 days. The rapid degradation of antibiotics and nutrient reformulation during BSFL conversion combined with composting indirectly altered manure bacterial communities, resulting in a lower abundance and richness of ARGs. The number of main antibiotic-resistant bacteria (e.g., Prevotella, Ruminococcus) decreased by 74.9 %, while their potential antagonistic bacteria (e.g., Bacillus, Pseudomonas) increased by 128.7 %. The number of antibiotic-resistant pathogenic bacteria (e.g., Selenomonas, Paenalcaligenes) decreased by 88.3 %, and the average number of ARGs carried by each human pathogenic bacterial genus declined by 55.8 %. BSF larvae gut microbiota (e.g., Clostridium butyricum, C. bornimense) could help reduce the risk of multidrug-resistant pathogens. These results provide insight into a novel approach to mitigate multidrug resistance from the animal industry in the environment by using insect technology combined with composting, in particular in light of the global "One Health" requirements.


Subject(s)
Composting , Diptera , Humans , Swine , Animals , Larva , Manure/microbiology , Anti-Bacterial Agents/pharmacology , Bacteria/genetics , Genes, Bacterial
10.
Genes (Basel) ; 13(12)2022 12 19.
Article in English | MEDLINE | ID: mdl-36553673

ABSTRACT

Feed efficiency (FE) is a very important trait affecting the economic benefits of pig breeding enterprises. Adipose tissue can modulate a variety of processes such as feed intake, energy metabolism and systemic physiological processes. However, the mechanism by which microRNAs (miRNAs) in adipose tissues regulate FE remains largely unknown. Therefore, this study aimed to screen potential miRNAs related to FE through miRNA sequencing. The miRNA profiles in porcine adipose tissues were obtained and 14 miRNAs were identified differentially expressed in adipose tissues of pigs with extreme differences in FE, of which 9 were down-regulated and 5 were up-regulated. GO and KEGG analyses indicated that these miRNAs were significantly related to lipid metabolism and these miRNAs modulated FE by regulating lipid metabolism. Subsequently, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) of five randomly selected DEMs was used to verify the reliability of miRNA-seq data. Furthermore, 39 differentially expressed target genes of these DEMs were obtained, and DEMs-target mRNA interaction networks were constructed. In addition, the most significantly down-regulated miRNAs, ssc-miR-122-5p and ssc-miR-192, might be the key miRNAs for FE. Our results reveal the mechanism by which adipose miRNAs regulate feed efficiency in pigs. This study provides a theoretical basis for the further study of swine feed efficiency improvement.


Subject(s)
MicroRNAs , Swine/genetics , Animals , MicroRNAs/genetics , MicroRNAs/metabolism , Reproducibility of Results , Lipid Metabolism , Phenotype , Adipose Tissue/metabolism
11.
Int J Biol Sci ; 18(16): 6176-6188, 2022.
Article in English | MEDLINE | ID: mdl-36439883

ABSTRACT

Mammals maintain a constant core body temperature through adaptive thermogenesis which includes shivering and non-shivering thermogenesis. Non-shivering thermogenesis relies primarily on mitochondrial uncoupling protein 1 (UCP1) in thermogenic fat (including brown and beige adipose tissue) to burn substrates, such as fatty acids (FAs), and convert chemical energy into heat. Lipid droplets (LDs), which are organelles that store lipids, are present in large numbers in thermogenic fat and are essential for adipose thermogenesis. Upon cold stimulation, LDs rapidly release FAs through autophagy or lipase-mediated lipolysis and rapidly translocate FAs into the mitochondria by interacting with mitochondria to burn and so promote thermogenesis. In addition, LD proteins promote the expression of UCP1 by activating the transcriptional activity of thermogenesis-related proteins. Here, the progress of research on the important role of LDs in thermogenesis is reviewed, mainly in terms of LD proteins, LD-organelle interactions, and LD autophagy (lipophagy). The emerging rationale for the involvement of LDs in each thermogenic pathway is described and the remaining unanswered questions in this field are highlighted.


Subject(s)
Lipid Droplets , Thermogenesis , Animals , Lipid Droplets/metabolism , Thermogenesis/genetics , Uncoupling Protein 1/metabolism , Adipose Tissue/metabolism , Mitochondria/metabolism , Mammals
12.
Ecotoxicol Environ Saf ; 243: 114006, 2022 Sep 15.
Article in English | MEDLINE | ID: mdl-36037632

ABSTRACT

Ammonia is one of the most important toxic metabolites in the intestine of animals. It can cause intestinal damage and associated intestinal diseases through different endogenous or exogenous stimuli. However, the definition of harmful ammonia concentration and the molecular mechanism of ammonia - induced intestinal epithelial injury remain unclear. In this study, we found that the viability of porcine IPEC-J2 intestinal epithelial cells significantly decreased with the increase of NH4Cl dose (20-80 mM). Ammonia (40 mM NH4Cl) increased the expression level of ammonia transporter RHCG and disrupted the intestinal barrier function of IPEC-J2 cells by reducing the expression levels of the tight junction molecules ZO-1 and Claudin-1. Ammonia caused elevated levels of ROS and apoptosis in IPEC-J2 cells. This was manifested by decreased activity of antioxidant enzymes SOD and GPx, decreased mitochondrial membrane potential, and increased cytoplasmic Ca2+ concentration. In addition, the expression levels of apoptosis-related molecules Caspase-9, Caspase-3, Fas, Caspase-8, p53 and Bax were increased, the expression level of anti-apoptotic molecule Bcl-2 was decreased. Moreover, the antioxidant NAC (N-acetyl-L-cysteamine) effectively alleviated ammonia-induced cytotoxicity, reduced ROS level, Ca2+ concentration, and the apoptosis of IPEC-J2 cells. The results suggest that ammonia-induced excess ROS triggered apoptosis through mitochondrial pathway, death receptor pathway and DNA damage. This study can provide reference and theoretical basis for the definition of harmful ammonia concentration in pig intestine and the effect and mechanism of ammonia on pig intestinal health.


Subject(s)
Ammonia , Antioxidants , Ammonia/metabolism , Ammonia/toxicity , Animals , Antioxidants/metabolism , Apoptosis , Cell Line , Epithelial Cells , Intestines , Reactive Oxygen Species/metabolism , Swine
13.
Int J Mol Sci ; 23(15)2022 Aug 05.
Article in English | MEDLINE | ID: mdl-35955862

ABSTRACT

The breakdown of lipid droplets (LDs) provides energy and contributes to the proliferation and migration of cancer cells. Recent studies have suggested that motility plays a key role in LD breakdown. However, the molecular mechanisms underlying LD motility were poorly characterized. In this study, we examined the function of microfilament-associated proteins 2 and 3 (ARP2 and ARP3) in regulating LDs' motility in Hela cells. ARP2/3 mediated the LDs' physical contact with F-actin and promoted the recruitment of Myosin Heavy Chain 9 (MYH9). MYH9 regulated the LD content by binding with LDs and ARP2/3. The number of LDs and TG content was increased after MYH9 interfered. The genes related to FA-related genes and neutral lipid synthesis-related genes were significantly increased (p < 0.05) when ARP2 and ARP3 were overexpressed. Bioinformatic analysis indicated that the high expression of ARP2/3 was associated with a poorer prognosis in cervical squamous cell carcinoma (CSCC). This study showed the effect of cytoskeletal filaments on LD metabolism in cancer cells.


Subject(s)
Actin-Related Protein 2-3 Complex/metabolism , Actins , Lipid Droplets , Actins/metabolism , Cytoskeletal Proteins/metabolism , Fatty Acids/metabolism , HeLa Cells , Humans , Lipid Droplets/metabolism , Lipid Metabolism , Microfilament Proteins/metabolism
14.
Life (Basel) ; 12(6)2022 Jun 10.
Article in English | MEDLINE | ID: mdl-35743903

ABSTRACT

The use of black soldier fly (BSF) larvae to recycle various organic materials while producing biomass for use as feed is well established. Variety selection is important from the perspective of application. In the current study, morphometric and life-history traits of a Wuhan-domesticated BSF colony (Wuhan strain) were compared to those of a 'selectively inbred' population (inbred strain, inbred for 10 generations). In terms of morphological characteristics, the results showed that both strains had dichoptic compound eyes, club-shaped antennae, blue halters, and blue-green metallic luster wings with a hexagon discal cell. In both strains, the body and wing length of female adults were slightly larger than those of male adults. The first four larval stages of the BSF occurred rapidly (1-12 days) with transitions across stages resulting in doubling of size for both populations. Selective inbreeding did not alter the life-history traits of the larval exuviate stage in terms of age, size, weight, and feed reduction rate. Overall egg production for the inbred strain was significantly higher (1.5 times greater) than the Wuhan strain. This is explained by increased adult emergence and individual oviposition performance. It was speculated that inbreeding improved the reproductive success of inbred adult female offspring and selection process steadied it. The findings indicate that selective inbreeding could enhance overall oviposition performance and provide a strategy to selectively breed BSF with high egg production for future applications.

15.
Commun Biol ; 5(1): 326, 2022 04 06.
Article in English | MEDLINE | ID: mdl-35388144

ABSTRACT

Hepatic steatosis is the main characteristic of some liver metabolism diseases. However, unclear molecular mechanism of hepatic steatosis impedes the therapy of this hepatic steatosis. Glutathione-S-transferase mu 2 (GSTM2), as a member of phase II drug metabolizing enzymes (DMEs), regulates cellular antioxidant and detoxificant. GSTM2 was highly up-regulated in hepatic steatosis tissues and high-fat diet (HFD) fed mice. Loss-of-function GSTM2 mouse model demonstrated that GSTM2 protected mice from excess fat accumulation. Mechanistically, GSTM2 interacted with ASK1 and suppressed its phosphorylation and the activation of subsequent downstream p38-JNK signalling. Moreover, GSTM2 overexpression in the liver effectively ameliorated hepatic lipid accumulation. Therefore, we identified GSTM2 as an important negative regulator in progression of hepatic steatosis via both its detoxification/antioxidant and inhibition of ASK1-p38/JNK signalling. This study showed potential therapeutic function of the DME in progression of hepatic steatosis.


Subject(s)
Non-alcoholic Fatty Liver Disease , Animals , Antioxidants , Glutathione Transferase/genetics , Mice , Non-alcoholic Fatty Liver Disease/metabolism
16.
Life Sci ; 299: 120540, 2022 Jun 15.
Article in English | MEDLINE | ID: mdl-35398332

ABSTRACT

AIMS: In mammals, heat stress (HS) from high-temperature environments has multiple adverse effects on the well-being of the organism. Brown adipose tissue (BAT) is a thermogenesis tissue that protects against obesity, and as an endocrine organ that regulates the systemic metabolism, but it is unclear how heat stress affects BAT in normal and obese subjects. Understanding the transcriptomic profiles and lipidomics of BAT upon heat exposure provides insights into the adaptive changes associated with this process. MATERIALS AND METHODS: We constructed heat treatment (40 °C, 4 h) models for normal and obese mice, observed the effect of heat treatment on interscapular BAT (iBAT) and performed an assay for iBAT with RNA-seq and lipidomics to compare transcriptional programs and lipid dynamics. KEY FINDINGS: In normal mice, heat treatment caused an iBAT damage by decreasing the expression of genes involved in thermogenesis, adipogenesis and lipid metabolism. Furthermore, HS disturbed the acyl-chain composition of triacylglycerols (TAGs) and glycerophospholipids (PEs, PCs and CLs), accelerated the production of cholesterol esters, and caused the formation of giant lipid droplets rich in cholesterol esters in iBAT. Unexpectedly, in obese mice, heat treatment had a smaller effect on iBAT by improving the composition of the saturated glycerolipids, PEs and PCs and increasing the proportion of oxidized lipid in lipid droplets. SIGNIFICANCE: Our findings proved lipid droplets participated in the regulation of lipid components of iBAT in normal and obese mice after heat treatment, which provided a new view for the understanding of the adaptation of iBAT to high-temperature environments.


Subject(s)
Adipose Tissue, Brown , Transcriptome , Adipose Tissue, Brown/metabolism , Animals , Cholesterol Esters/metabolism , Humans , Lipid Droplets , Lipidomics , Mammals , Mice , Mice, Obese , Obesity/metabolism , Thermogenesis
17.
Nanomaterials (Basel) ; 12(4)2022 Feb 17.
Article in English | MEDLINE | ID: mdl-35215001

ABSTRACT

Lipid droplets (LDs) play an important role in the regulation of cellular stress. This suggests LDs can be applied as safe and effective biomaterials to alleviate cellular stress and lipotoxicity. Here, we constructed a convenient method to generate stable and pure artificial lipid droplets (aLDs). aLDs can maintain their biological function by incubating LD-associated proteins or organelles in vitro. It was validated that perilipin-coated aLDs could be uptaken by cells, significantly reducing hydrogen peroxide-induced reactive oxidative species (ROS) and alleviating cellular lipotoxicity caused by excess fatty acid. Our work demonstrated a direct role of LDs in regulating cellular stress levels, providing methods and potential value for future research and medical applications of LDs.

18.
Mol Biotechnol ; 64(5): 473-481, 2022 May.
Article in English | MEDLINE | ID: mdl-34822105

ABSTRACT

Transgenic promoter systems are of great interest for their potential use in gene therapy or production due to their high activity, long term, and cell specificity. Here, in order to obtain promoters with high activity and expressed specifically in skeletal muscle, the MYOD1, MYF5, and MCK were selected as the candidate genes. The truncated promoters were amplified and their activity was verified through dual-luciferase reporter gene test. We used genetic engineering techniques to improve promoter activity by tandemly linking enhancers and promoters or two promoters. Furthermore, synthetic promoter was the most active when two eMCK enhancers and pMCK promoter were cascaded. To improve the tissue specificity of the promoter, the seed region of translational repressor miR-208a was inserted into the downstream of the promoter (pGL3-2eMCK-pMCK-T208-mCherry-EGFP). The results showed that the expression level of target genes decreased significantly (P < 0.05) in myocardium rather than in skeletal muscle. The results of in vivo transfection indicated that tandem transcriptional regulatory elements can increase promoter activity in mice. This work laid the foundation for future research on genetically modified pigs.


Subject(s)
MicroRNAs , Regulatory Sequences, Nucleic Acid , Animals , Enhancer Elements, Genetic , Genes, Reporter , Mice , MicroRNAs/genetics , MicroRNAs/metabolism , Muscle, Skeletal/metabolism , Promoter Regions, Genetic , Swine/genetics
19.
Cell Death Dis ; 12(12): 1102, 2021 11 23.
Article in English | MEDLINE | ID: mdl-34815388

ABSTRACT

Lipid droplet (LD), a multi-functional organelle, is found in most eukaryotic cells. LDs participate in the regulation of many cellular processes including proliferation, stress, and apoptosis. Previous studies showed the athlete's paradox that trained athletes accumulate LDs in their skeletal muscle. However, the impact of LDs on skeletal muscle and myogenesis is not clear. We discovered that C2C12 myoblast cells containing more LDs formed more multinucleated muscle fibers. We also discovered that LDs promoted cell migration and fusion by promoting actin-filaments remodeling. Mechanistically, two LD-proteins, Acyl-CoA synthetase long chain family member 3 (ACSL3) and lysophosphatidylcholine acyltransferase 1 (LPCAT1), medicated the recruitment of actinin proteins which contributed to actin-filaments formation on the surface of LDs. During remodeling, the actinin proteins on LDs surface translocated to actin-filaments via ARF1/COPI vesicles. Our study demonstrate LDs contribute to cell differentiation, which lead to new insight into the LD function.


Subject(s)
Lipid Droplets/metabolism , Cell Differentiation , Humans , Transfection
20.
Front Immunol ; 12: 696148, 2021.
Article in English | MEDLINE | ID: mdl-34603279

ABSTRACT

As the first line of defense against intestinal bacteria and toxins, intestinal epithelial cells are always exposed to bacteria or lipopolysaccharide (LPS), whereas pathogenic bacteria or LPS can cause intestinal epithelial cell damage. Previous studies have shown that konjac mannan oligosaccharides (KMOS) have a positive effect on maintaining intestinal integrity, and Bacillus subtilis (BS) can promote the barrier effect of the intestine. However, it is still unknown whether KMOS and BS have a synergistic protective effect on the intestines. In this study, we used the LPS-induced Caco-2 cell injury model and mouse intestinal injury model to study the synergistic effects of KMOS and BS. Compared with KMOS or BS alone, co-treatment with KMOS and BS significantly enhanced the activity and antioxidant capacity of Caco-2 cell, protected mouse liver and ileum from LPS-induced oxidative damage, and repaired tight junction and mucus barrier damage by up-regulating the expression of Claudin-1, ZO-1 and MUC-2. Our results demonstrate that the combination of KMOS and BS has a synergistic repair effect on inflammatory and oxidative damage of Caco-2 cells and aIIeviates LPS-induced acute intestinal injury in mice.


Subject(s)
Bacillus subtilis/physiology , Epithelial Cells/drug effects , Epithelial Cells/microbiology , Ileal Diseases/prevention & control , Intestinal Mucosa/drug effects , Intestinal Mucosa/microbiology , Mannans/pharmacology , Probiotics , Animals , Caco-2 Cells , Cell Survival/drug effects , Chemical and Drug Induced Liver Injury/metabolism , Chemical and Drug Induced Liver Injury/microbiology , Chemical and Drug Induced Liver Injury/pathology , Chemical and Drug Induced Liver Injury/prevention & control , Disease Models, Animal , Epithelial Cells/metabolism , Epithelial Cells/pathology , Female , Humans , Ileal Diseases/metabolism , Ileal Diseases/microbiology , Ileal Diseases/pathology , Intestinal Mucosa/metabolism , Intestinal Mucosa/pathology , Lipopolysaccharides , Mice , Oxidative Stress/drug effects , Permeability , Tight Junctions/drug effects , Tight Junctions/metabolism , Tight Junctions/microbiology , Tight Junctions/pathology
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