Synthesis-related Genes Research Articles

Regulation of Rice Grain Quality by Exogenous Kinetin During Grain-Filling Period

Cytokinins (CKs) play important functions in plant growth and development and in response to adversity stress. However, little is known about the role CK plays in rice grain quality. We hypothesized that exogenous cytokinins could improve rice grain quality by regulating physiological traits and genes related to starch synthesis. Therefore, we exogenously applied different concentrations of kinetin (KT), an exogenous CK, during the grain-filling period. Our results show that all the different concentrations of exogenous KT treatments resulted in a significant increase in thousand-grain weight. In particular, chalkiness and chalky grain rate were significantly reduced, and gel consistency (GC) content and alkali spreading value (ASV) were significantly increased in 10−8 M KT treatment. Meanwhile, the exogenous application of 10−8 M KT positively affected the transcription of some starch synthesis-related genes, which was in contrast to the 10−5 M KT treatment. In conclusion, the exogenous application of appropriate concentrations of KT during the grain-filling period can ultimately affect rice grain quality by regulating the changes in the relevant indicators, such as appearance quality (AQ) and eating and cooking qualities (ECQ).

DOP123 Linking microbial genes to mucosal metabolites uncovers host-microbial interactions during 5-ASA therapy for ulcerative colitis

Abstract Background The severity of ulcerative colitis (UC) depends on the state of the colon mucosa, where achieving mucosal remission markedly improves patient prognosis and quality of life. The study aimed to characterize essential mucosal metabolites and microbial gene involved in UC remission, and to elucidate the mechanisms by which specific metabolites regulate intestinal homeostasis. Methods An integrated metagenomic and metabolomic analysis was conducted to elucidate the correlation between disease-associated microbes and host mucosal metabolites. Participants included non-UC controls and individuals with UC, comprising the responding group (UCR) and ineffective group (UCA) in 5-ASA treatment. The differential metabolite was further investigated using a DSS-induced colitis mouse model, followed by in vitro experiments to explore the mechanisms. Results The levels of mucosal secondary bile acids were found to have a significant negative association with intestinal inflammation during 5-ASA treatment. Integrated analysis of metagenomics and metabolomics revealed that host mucosal bile acid metabolites were negatively associated with the bile acid synthesis-related genes cbh in microbial genomes. Administration of the secondary bile acid mitigated mucosal inflammation in a colitis model by decreasing the proportion of Th17 and pathogenic Th17 cells. This secondary bile acid functions as an immunomodulatory metabolite, suppressing glycolysis in Th17-polarizing T cells to inhibit the Th17 cell response and modulating the gut microbiota. Conclusion These findings suggest that alterations in microbiome-metabolome interactions are critical for maintaining mucosal homeostasis during UC remission. The specific bile acid acts as an immunomodulatory metabolite, and may represent a potential therapeutic strategy for UC.

Oridonin inhibits SGIV infection by regulating glycolipid metabolism and inflammatory response.

Singapore grouper iridovirus (SGIV) is a significant infectious disease in the grouper aquaculture industry. Currently, there is no effective drug available to prevent or treat SGIV. Oridonin (Ori) is a naturally occurring compound derived from Rabdosia rubescens, exhibiting various biological activities, including anti-tumor, anti-inflammatory, and antioxidant properties. In this study, we examined the anti-SGIV activity of Ori and its potential mechanism of action in vitro. The study results indicate that Ori effectively inhibits SGIV infection at various concentrations. Further studies reveal that Ori inhibits the formation of lipid droplets induced by SGIV infection. Additionally, Ori suppresses the SGIV-induced up-regulation of fatty acid synthesis-related genes (SREBP1, ACC1, SCD1, FASN) and glycolysis-related genes (GLUT1, GLUT2, HK2, PDHX). The mTOR pathway plays a crucial role in regulating glycolipid metabolism. Our findings indicate that Ori suppresses the phosphorylation of AKT and mTOR proteins. Further research revealed that the activation or inhibition of mTOR significantly impacts SGIV protein production and the expression of genes related to glycolipid metabolism. In addition, Ori effectively inhibits the up-regulation of NLRP3, ASC, Caspase-1, and pro-inflammatory cytokines induced by SGIV infection. In conclusion, our experimental findings indicate that Ori effectively inhibits SGIV infection by regulating glycolipid metabolism through the AKT/mTOR pathway and by suppressing the inflammatory responses triggered by SGIV infection.

Sex and age effects on chronic inflammatory pain development, maintenance, and resolution in Wistar rats.

Millions of Americans live with chronic inflammatory pain conditions, and the prevalence of these conditions increases with age and is higher in females. Still, it is poorly understood how sex, age and peripheral gene expression affect the trajectory of chronic inflammatory pain conditions. We used the inflammatory agent, Complete Freund's Adjuvant (CFA), to systematically test sex and age effects on mechanical and thermal sensitivity in adolescent and adult male and female Wistar rats over 3 weeks (Experiment 1 [onset]) or 11 weeks (Experiment 2 [recovery]). We report that relative to male CFA rats, CFA females are mechanically hypersensitive at all time points and thermally hypersensitive at later time points (long-term during maintenance and recovery). Also, within CFA rats, more adult males (90%) achieved behavioral recovery at 11 weeks, relative to adolescent males (70%), adult females (70%) and adolescent females (50%). Behavioral recovery was most highly correlated with thermal nociception scores in most groups. Among adult males, significant positive correlations were seen between mechanical and thermal nociception scores and between scores in the CFA-injected and non-injected paws. In paw tissue from a subset of rats from experiment 2, we also report increased transient receptor potential cation channel subfamily V member 1 (TRPV1) gene expression in adult CFA but not adolescent CFA rats, and increased pro- and anti-inflammatory, and triglyceride synthesis-related gene expression in all CFA rats. Our results demonstrate apparent sex and age differences in the trajectory of chronic inflammatory pain-related behavior and gene expression in the affected paw of rats. PERSPECTIVE: This article highlights sex and age differences in the trajectory of chronic inflammatory pain and possible peripheral mechanisms driving recovery in Wistar rats. This data could be used to better understand recovery patterns in patients with this type of pain and provides a starting point for assessment of novel treatments.

Integrated Genetic Diversity and Multi-Omics Analysis of Colour Formation in Safflower.

Safflower (Carthamus tinctorius L.) is a medicinal and edible cash crop that is widely cultivated worldwide. However, the genetic diversity of safflower germplasm resources and the reasons for the variations in safflower flower colour remain unclear. In this study, we used a combination of agronomic traits and Indel markers to assess the genetic diversity of 614 safflower germplasm resources. The results showed that most of the evaluated agronomic traits had high variability. The mean values of the Shannon's information index (I) and polymorphism information content (PIC) in 50 pairs of Indel markers were 0.551 and 0.296, respectively. The population structure, neighbour-joining phylogeny, and principal coordinate analyses classified all genotypes into four subgroups, and 214 safflower core germplasms were constructed. Multiple analyses of genetic diversity parameters, range conformity, and the percentage of variance difference showed that the core germplasm did not differ significantly and could represent the original germplasm better. Transcriptome and metabolome analyses revealed that flavonoid synthesis-related genes, including CHS, F3H, ANS, and BZ1, were differentially expressed in different coloured safflowers. Most significantly, different genes and metabolite compounds in white safflowers were enriched upstream from the phenylpropanoid metabolic pathway to the production of naringenin, whereas those in red safflowers were concentrated in the downstream pathway from eriodictyol. Meanwhile, the preliminary quantification of anthocyanins and carotenoids extracted from red, orange, and white types of safflower showed that the level of both anthocyanins and carotenoids were highest in red types. This work provides new insights into the formation of different safflower flower colours and in the conservation and management of safflower germplasm.

Rubus idaeus RiACS1 Gene Is Involved in Ethylene Synthesis and Accelerates Fruit Ripening in Solanum lycopersicum

Raspberry is a berry whose fruit is not tolerant to storage; breeding varieties with extended storage time and high comprehensive quality are significant for raspberries in cold regions. 1-Aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) is a limiting enzyme in the ethylene synthesis process, which plays essential roles in fruit ripening and softening in plants. In this study, the RiACS1 gene in raspberry (Rubus idaeus L.) variety ‘Polka’ was cloned. The RiACS1 gene overexpression vector was constructed and transformed into tomato plants using the Agrobacterium tumefaciens infection method to verify its function in their reproductive development. The RiACS1 gene, with a total length of 1476 bp, encoded a protein with 491 amino acids. The subcellular localization analysis of the RiACS1 protein in the tobacco transient expression system revealed that the RiACS1-GFP fusion protein was mainly located in the nucleus. Compared with the control, the flowering time and fruit color turning time of transgenic strains were advanced, and the fruit hardness was reduced. Overexpression of RiACS1 increased the activity of ACC synthase, ethylene release rate, and respiration rate during the transchromic phase. It changed the substance content, increased the content of vitamin C and anthocyanin in the fruit ripening process, and decreased the content of chlorophyll and titrable acid at the maturity stage. In addition, RiACS1 increased the relative expression levels of ethylene synthesis-related genes such as SlACS4, SlACO3, and SlACO1 in the fruit ripening process, while it decreased the expression levels of SlACS2 at the maturity stage. These results suggested that the RiACS1 gene could promote early flowering and fruit ripening in tomato plants. This study provided a basis for further modifying raspberry varieties using molecular biology techniques.

Porcine-Derived Chondroitin Sulfate Sodium Alleviates Osteoarthritis in HTB-94 Cells and MIA-Induced SD Rat Models.

Osteoarthritis (OA) is a chronic disease characterized by cartilage degradation, leading to bone friction, inflammation, stiffness, pain, and reduced mobility. This study investigates the therapeutic effects of porcine-derived chondroitin sulfate sodium (CS) on OA symptoms at both cellular and animal levels. In vitro study, HTB-94 chondrocytes were treated with inflammatory stimuli and CS (10, 50, 100, and 200 μg/mL) to assess the release of inflammatory mediators and the expression of genes and proteins related to cartilage synthesis and degradation. In vivo study, an MIA-induced OA rat model was used, and CS (62, 124, and 248 mg/kg b.w.) was orally administered for 4 weeks. Key parameters, such as exercise capacity, micro-CT, histological evaluation of joint tissues, serum inflammatory markers, and the expression of mRNA and proteins (inflammatory, cartilage synthesis and degradation, and apoptosis markers), were analyzed. Porcine-derived CS significantly reduced PGE2, NO, and extracellular matrix degradation marker (COMP and CTX-II) levels and increased the expression of cartilage synthesis-related genes and proteins in both HTB-94 cells and the MIA-induced rats. Additionally, CS modulated cartilage degradation pathways and notably inhibited apoptosis in vivo. The effects of porcine CS were comparable to the NSAID ibuprofen, demonstrating its potential as an anti-inflammatory and chondroprotective agent for OA management and dietary supplementation.

Gga-miR-6634-5p Affects the proliferation and steroid hormone secretion of chicken (Gallus Gallus) granulosa cells by targeting MMP16.

MiRNAs are typically reported to play a negative regulatory role in post-transcriptional expression of target genes and are widely involved in a variety of biological processes such as growth, metabolism and reproduction. However, research on the role of miRNAs in the ovulation process of chicken ovaries is still insufficient compared to that in mammals. Here, we investigated the regulatory mechanisms of gga-miR-6634-5p in the growth and steroid hormone secretion of chicken granulosa cells (GCs) by targeting MMP16. We found that gga-miR-6634-5p significantly down-regulated the mRNA levels of proliferation-related genes (CCND1, CDK1, and CDK6), decreased cell viability, the number of EdU-labelled positive cells, and the percentage of S-phase cells, as analysed by quantitative real time PCR (qRT-PCR), cell counting kit-8 (CCK-8), 5-ethynyl-2'deoxyuridine (EdU) and flow cytometry analyses (P < 0.01 or P < 0.05). qRT-PCR and enzyme-linked immunosorbent assay (ELISA) results demonstrated that gga-miR-6634-5p up-regulated the expression of steroid synthesis-related genes (CYP19A1, 3β-HSD, StAR and FSHR) (P < 0.01 or P < 0.05), as well as the secretion of estradiol (E2) and progesterone (P4) (P < 0.01 or P < 0.05). Furthermore, we found that MMP16 protein and gene expression can be down-regulated by gga-miR-6634-5p and demonstrated that MMP16 is a target gene of gga-miR-6634-5p by dual luciferase reporter assay (P < 0.05). In addition, we found that MMP16 stimulated the proliferation of GCs, significantly inhibited the expression of steroid synthesis related genes (CYP19A1, StAR, 3β-HSD and FSHR), and decreased the secretion of E2 and P4 (P < 0.01 or P < 0.05), which was consistent with the inhibitory effect of gga-miR-6634-5p. It was further found by functional enrichment analysis, qRT-PCR, western blot (WB) and ELISA that MMP16 may play a regulatory role in GCs proliferation as well as steroid hormone secretion through the mTOR signaling pathway and PPAR signaling pathway. Therefore, this study demonstrates that gga-miR-6634-5p modulates the proliferation of chicken GCs and the secretion of steroid hormones by targeting MMP16, which may contribute to a better understanding of the functional mechanisms of miRNAs in the ovarian development of laying hens.

Pachymic acid promotes brown/beige adipocyte differentiation and lipid metabolism in preadipocytes 3T3-L1 MBX.

To investigate the effect of pachymic acid on brown/beige adipocyte differentiation and lipid metabolism in preadipocytes 3T3-L1 MBX. The brown cocktail method was employed to induce 3T3-L1 MBX cells to differentiate into beige adipocytes. The impact of pachymic acid on the viability of 3T3-L1 MBX preadipocytes was evaluated using the CCK-8 assay. The formation of lipid droplets following treatment with pachymic acid was observed through oil red O staining, and the content of lipids in differentiated cells was determined. The expression levels of key browning genes, including uncoupling protein (Ucp) 1, the peroxisome proliferation-activating receptor gamma coactivator (Pgc)-1α, and the transcription factor containing PR domain 16 (Prdm16) were detected by quantitative reverse transcription polymerase chain reaction. The expression of sterol regulatory element binding protein (Srebp) 1c, acetyl-CoA carboxylase (Acc), fatty acid synthetase (Fas), and steroid-sensitive lipase (Hsl), fatty triglyceride hydrolase (Atgl), and carnitine palmitoyl transferase (Cpt) 1 of lipolysis-related genes were also examined. The 3T3-L1 MBX was induced in vitro to form beige adipocytes with high expression of key browning genes, including Ucp1, Pgc-1α, Prdm16, and beige adipose-marker genes, including Cd137, Tbx1, and Tmem26. The concentration range of 0-80 μM pachymic acid was non-cytotoxic to 3T3-L1 MBX. Pachymic acid treatment significantly inhibited the differentiation of 3T3-L1 MBX, resulting in a notable decrease in lipid accumulation content (P<0.01). Additionally, there was a marked increase in the expression of key browning genes and their proteins, such as Ucp1, Pgc-1α, and Prdm16, while the expressions of fat synthesis-related genes Srebp1c, Acc and Fas were significantly decreased (all P<0.05). The expressions of lipolysis-related genes, including Hsl, Atgl, and Cpt1, were significantly increased (all P<0.05). Besides, treating with 20 μmol/L pachymic acid showed the most pronounced effect. Pachymic acid can inhibit fat synthesis and promote lipid decomposition by regulating the brown formation and lipid differentiation of 3T3-L1 MBX preadipocytes.

Zinc finger protein rotund is essential for wings and ovarian development by regulating lipid homeostasis in Locusta migratoria.

Cys2-His2-type zinc finger (C2H2-ZF) proteins are involved in diverse biological processes. In insects, the wing and ovarian development is crucial for reproduction and evolution, yet the physiological roles of C2H2-ZF proteins in these processes remain underexplored. Here, RNA-seq analyses identified C2H2-ZF protein genes that were differentially expressed during wing formation in Locusta migratoria. Among these, the gene encoding a C2H2-ZF protein Rotund (Rn) was highly expressed in the wing pads of fourth- and fifth-instar nymphs. RNA interference mediated knockdown of LmRn in nymph stages resulted in pronounced abnormalities with curled wings and reduced wing area. LmRn knockdown led to reduced expression of lipid transport-related genes during wing morphogenesis, significantly decreased triglyceride (TG) level. In addition, we also find that LmRn knockdown impaired ovarian development and oocyte maturation in female adults, with decreased expression levels of lipid synthesis-related genes, and vitellogenin genes (VgA, and VgB) in the fat body. Meanwhile, the number of lipid droplets, TG content, and protein levels in the ovaries were significantly decreased after LmRn was silenced. Together, our findings reveal that LmRn is essential for wing and ovarian development by regulating lipid homeostasis in locusts, offering a potential target for insect pest management.

Transcriptomic Analysis of Wheat Under Multi LED Light Conditions.

Light is a vital environmental cue that profoundly influences the development of plants. LED lighting offers significant advantages in controlled growth environments over fluorescent lighting. Under monochromatic blue LED light, wheat plants exhibited reduced stature, accelerated spike development, and a shortened flowering period with increased blue light intensity promoting an earlier heading date. In this study, we conducted a comprehensive transcriptome analysis to investigate the molecular mechanisms underlying wheat plants' response to varying light conditions. We identified 34 types of transcription factors (TFs) and highlighted the dynamic changes of key families such as WRKY, AP2/ERF, MYB, bHLH, and NAC, which play crucial roles in light-induced gene regulation. Additionally, this study revealed differential effects of blue and red light on the expression levels of genes related to hormones such as cytokinin (CK) and salicylic acid (SA) synthesis as well as significant changes in pathways such as flavonoid biosynthesis, circadian rhythms, chlorophyll synthesis, and flowering. Particularly, blue light upregulated genes involved in chlorophyll synthesis, contrasting with the downregulation observed under red light. Furthermore, blue light enhanced the expression of anthocyanin synthesis-related genes, such as CHS, underscoring its role in promoting anthocyanin accumulation. These findings provide valuable insights into how light quality impacts crop growth and development.

Enhancing the nutritional profile of rice by targeting starch branching enzymes using CRISPR/Cas9

Rice is a fundamental staple in many Asian countries; however, excessive consumption can lead to significant health concerns, including diabetes. One effective strategy to mitigate these concerns is to increase the amylose content in rice, which enhances its resistant starch (RS) levels. Higher RS not only improves the nutritional profile of rice but also positively impacts its cooking qualities, offering various health benefits. Recent research highlights the role of dietary fibers like RS in modulating gut microbiota composition, presenting a promising approach for addressing non-communicable diseases. RS enhances the fermentation activity of gut microbiota, leading to production of beneficial metabolites that support gut barrier function, exhibit anti-inflammatory properties and influence metabolic pathways related to obesity and diabetes. This multifaceted impact on chronic disease outcomes emphasizes the need for rice varieties with increased amylose and consequently higher RS levels, to meet consumer nutritional demands. CRISPR/Cas9, a powerful genome editing tool, allows precise modifications of the targeted genes. This technology can effectively edit starch synthesis-related genes in rice to enhance starch content. This review focuses on the application of CRISPR/Cas9 in increasing RS content in rice and the potential health benefits it could provide to populations that rely on rice as a dietary staple. By integrating genetic innovation with nutritional science, healthier rice varieties can be developed, that align with the dietary needs of consumers.

Effects of high-fat diet on the morphology and function of the thyroid gland and glycolipid metabolism in thyroid clock gene Bmal1 knockout mice

Objective: To investigate the changes in thyroid morphology and function in mice with thyroid-specific knockout of the clock gene Bmal1 under high-fat diet (HFD), and to examine its effects on glycolipid metabolism in mice. Methods: Construct a mouse model with specific knockout of the Bmal1 gene in the thyroid (T-Bmal1-/-) (knockout group, n=10), and use Bmal1flox/flox mice without thyroid peroxidase-cyclization recombination enzyme (T-Bmal1+/+) as the control group (non-knockout group, n=10). The mice were fed until 6 weeks (body weight 20-23 g), and then use the random number table method to evenly divide each group of mice into two subgroups. They were then fed either with normal diet (ND) or (HFD), resulting in four final groups: ND non-knockout group, HFD non-knockout group, ND knockout group, and HFD knockout group, with 5 mice in each group. From the 6th week onwards, the body weights of the mice were measured for 10 consecutive weeks. On the first day of the 14th week, the intraperitoneal glucose tolerance test (IPGTT) was conducted, and on the first day of the 15th week, the intraperitoneal insulin tolerance test (IPITT) was performed. On the first day of the 16th week after the mice were born, blood samples (0.2 ml) were taken from the eyes under anesthesia at 7∶00, 13:00, 19:00, and 1∶00 the next day, and the thyroid, liver, kidneys, spleen, inguinal white adipose tissue, and scapular brown adipose tissue were rapidly removed. Thyroid tissue morphology, thyroid function indexes, thyroid clock genes and thyroid hormone synthesis and secretion related genes expression, body weight, IPGTT and IPITT area under the curve (AUC) and lipid levels were detected and compared among all groups. Results: There was no significant difference in the number of thyroid follicles, the area of thyroid follicles and the height of thyroid follicle cavity between ND knockout group and ND non-knockout group (all P>0.05), but the height of thyroid follicle cavity in HFD knockout group was lower than that in HFD knockout group [(25.8±1.6) vs (54.4±9.5) μm, P=0.002]. The level of serum T4 in knockout group after ND or HFD feeding was higher than that in non-knockout group (all P<0.05). There was no significant difference in the mRNA expression level of sodium iodine transporter between ND knockout group and ND non-knockout group (P=0.550), but the mRNA expression level of sodium iodine transporter in HFD knockout group was higher than that in HFD non-knockout group [0.67±0.27 vs 0.20±0.09, P=0.006].There was no significant difference in weight gain and groin white fat weight between ND knockout group and ND non-knockout group (both P<0.05), but weight gain and groin white fat tissue weight in HFD knockout group were lower than those in HFD knockout group (both P<0.05). The IPGTT results showed no statistically significant difference in AUC between the ND knockout and ND non-knockout groups (P=0.226), but the HFD knockout group had a smaller AUC than the HFD non-knockout group (P=0.008). The IPITT revealed that the AUC of the ND knockout group was smaller than that of the ND non-knockout group (P=0.047). Lipid profiles were similar between the ND knockout and ND non-knockout groups (all P>0.05), but the HFD knockout group had lower levels of total cholesterol and high-density lipoprotein cholesterol than the HFD non-knockout group (both P<0.05). Conclusion: Thyroid-specific knockout of the clock gene Bmal1 can affect thyroid hormone levels and glycolipid metabolism in mice, with more pronounced effects after HFD feeding.

Dynamic changes in the transcriptome and metabolome of pig ovaries across developmental stages and gestation

BackgroundThe ovary is a central organ in the reproductive system that produces oocytes and synthesizes and secretes steroid hormones. Healthy development and regular cyclical change in the ovary is crucial for regulating reproductive processes. However, the key genes and metabolites that regulate ovarian development and pregnancy have not been fully elucidated. This study conducted high-throughput RNA sequencing and untargeted metabolite profiling of the ovarian tissues from Chenghua pigs at four stages, including postnatal day 3 (D3), puberty at the age of about 125 days (Pub), sexual maturity at the age of about 365 days (Y1), and 105 days after pregnancy at the age of about 360 days (Pre).ResultsA total of 9,264 and 1,593 differentially expressed genes (DEGs) were identified during ovarian development and pregnancy. Several key genes involved in ovarian development, including SQLE, HMGCS1, MSMO1, SCARB1, CYP11A1, HSD3B1, HSD17B1, and SERPINE1 were identified. Similarly, LUM, FN1, PLAUR, SELP, SDC1, and VCAN were considered to be associated with pregnancy maintenance. Overexpression of HSD17B1 in granulosa cells significantly upregulated estrogen synthesis-related genes (HSD3B1, CYP11A1, and STAR); meanwhile, overexpression of PLAUR promotes granulosa cell proliferation. Furthermore, 66, 24, 77, and 7 differentially expressed miRNAs (DEMis) were found, leading to the selection of key miRNAs such as ssc-miR-206, ssc-miR-107, ssc-miR-429, ssc-miR-210, and ssc-miR-133a-3p by differential miRNA-targeted mRNA interaction network; meanwhile, ssc-miR-133a-3p was validated to have a targeting relationship with KCNA1 by dual-luciferase reporter systems assay. At the metabolic levels, androstenedione, 17a-hydroxyprogesterone, dehydroepiandrosterone, and progesterone were identified, with their synthesis regulated by these DEGs in the ovarian steroidogenesis pathway. Furthermore, treatment of cells with androstenedione upregulated the expression of HSD3B1, CYP11A1, and STAR.ConclusionsThis study revealed the dynamic changes in the transcriptome and metabolome of pig ovaries across developmental stages and gestation, indicating that it may provide new theoretical insights for improving sow fertility.

Exosomes Derived from Adipose Mesenhymal Stem Cells Ameliorate Lipid Metabolism Disturbances Following Liver Ischemia-Reperfusion Injury in Miniature Swine.

The liver plays a crucial role in regulating lipid metabolism. Our study examined the impact of Exosomes derived from adipose mesenchymal stem cells (ADSCs-Exo) on lipid metabolism following liver ischemia-reperfusion injury (IRI) combined with partial hepatectomy. We developed a miniature swine model for a minimally invasive hemi-hepatectomy combined with liver IRI. In this study, we administered PBS, ADSCs-Exo, and adipose-derived stem cells (ADSCs) individually through the portal vein. Before and after surgery, we evaluated various factors including hepatocyte ultrastructure, lipid accumulation in liver tissue, and expression levels of genes and proteins associated with lipid metabolism. In addition, we measured serum and liver tissue levels of high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides (TG), and total cholesterol (CHOL). TEM and oil red O stain indicated a significant reduction in liver steatosis following ADSCs-Exo treatment, which also elevated serum levels of HDL, LDL, TG, and CHOL. Additionally, ADSCs-Exo have been shown to significantly decrease serum concentrations of HDL, LDL, TG, and CHOL in the liver (p < 0.05). Finally, ADSCs-Exo significantly downregulated lipid synthesis-related genes and proteins, including SREBP-1, SREBP-2, ACC1, and FASN (p < 0.05), while upregulating lipid catabolism-related genes and proteins, such as PPAR-α and ACOX1 (p < 0.05). ADSCs-Exo as a cell-free therapy highlights its therapeutic potential in hepatic lipid metabolism abnormalities.

The mechanism of DEHP-induced lipid accumulation in liver of female zebrafish

Diethylhexyl phthalate (DEHP) is a typical environmental pollutant and poses a potential threat to organisms by disrupting the lipid metabolism. This study found that DEHP at environmental concentrations, led to lipid accumulation in female zebrafish, as indicated by significant increases in the content of total cholesterol, triglycerides and the lipid droplets, in a concentration-dependent manner. However, how DEHP induces the lipid accumulation remains poorly understood. Our results demonstrated that DEHP up-regulated the expression of fat synthesis related-genes fas, acc, acs, elvol6, scd and dgat1, and increased the enzymatic activity of fatty acid synthase and acetyl-CoA carboxylase. Furthermore, the expression of several key transcription factors that regulate fat synthesis was detected, among which active sterol regulatory element-binding protein-1 (SREBP-1) was significantly increased. When active SREBP-1 was inhibited with specific inhibitor or knocked down by transient transfection, the expression of lipid synthesis-related genes was significantly decreased in DEHP group, indicating that DEHP disrupted the lipid synthesis via SREBP-1 pathway. Additionally, molecular docking revealed direct interaction sites between DEHP and SREBP-1. Our findings revealed that DEHP could directly activate SREBP-1-mediated lipid synthesis, providing theoretical basis for DEHP threatening biological health.

Integrated omics profiles for exploring the potential mechanism underlying aroma formation in the terpenoid-rich aromatic plant Opisthopappus taihangensis and the bioactivity of its leaf essential oil

Opisthopappus taihangensis (Anthemideae, Asteraceae), which is rich in bioactive components, produces flowers and leaves with robust fragrances. In this study, we conducted comprehensive metabolomic and transcriptomic analyses to identify changes in terpenoid metabolites and associated gene expression across various O. taihangensis tissues (leaves, buds, and inflorescences at the exposure, initial-bloom, and full-bloom stages). We identified 1,370 metabolites using headspace solid-phase micro-extraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS), 308 of which were terpenoid metabolites. The expression of terpenoid synthesis-related genes was relatively consistent with the synthesis of terpenoid metabolites at each examined developmental stage. An analysis of gene networks governing terpenoid synthesis revealed that MCT genes (OtMCT1, OtMCT2, and OtMCT3), TPS genes (OtTPS5, OtTPS9, and OtTPS10), and OtISPS1 may be crucial for synthesizing specific metabolites in different tissues. Additionally, the essential oil extracted from leaves by water distillation showed thatthujone and camphor were the predominant components. The considerable antioxidant activity of the leaf essential oil was comparable to that of vitamin C (16 μg/mL). Notably, its antimicrobial effects against Staphylococcus aureus and Escherichia coli growth were greater than those of ampicillin and vancomycin at the same concentrations. Scanning electron microscopy images indicated that the leaf essential oil significantly disrupted bacterial cell structures. This study thoroughly analyzed the network of genes regulating terpenoid metabolites in different O. taihangensis tissues, and elucidated the antioxidant and antibacterial potential of the leaf essential oil, thus providing valuable insights for breeding, molecular characterization, and the potential application of O. taihangensis in developing useful essential oil-based natural products.

Baseline sensitivity and physiological characteristics of natural product hinokitiol against Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum, a pathogenic fungus of oilseed rape, poses a severe threat to the oilseed rapeseed industry. In this study, we evaluated the potential of the natural compound hinokitiol against S. sclerotiorum by determining its biological activity and physiological characteristics. Our results showed that hinokitiol strongly inhibited the hyphae expansion of S. sclerotiorum, and its effective concentration of hyphae growing inhibition by 50% (EC50) against 103 S. sclerotiorum strains varied from 0.36 to 3.45 μg/mL, with an average of 1.23 μg/mL. Hinokitiol possessed better protective efficacy than therapeutic effects, and it exhibited no cross-resistance between carbendazim. After treatment with hinokitiol, many vesicular protrusions developed on the mycelium with rough surface and thickened cell wall. Moreover, the cell membrane permeability and glycerol content increased, while the oxalic acid declined after hinokitiol treatment. In addition, hinokitiol induced membrane lipid peroxidation and improved the production of reactive oxygen species (ROS) in S. sclerotiorum. Importantly, real-time quantitative polymerase chain reaction showed that cell wall and ROS synthesis-related genes were significantly up-regulated after hinokitiol treatment. This study revealed that hinokitiol has good biological activity against S. sclerotiorum and could be considered as an alternative bio-fungicide for the resistance management in controlling sclerotinia stem rot infected by S. sclerotiorum. These investigations provided new insights into understanding the toxic action of hinokitiol against pathogenic fungi. © 2024 Society of Chemical Industry.

Physiological function of hydrophobin Hydph16 in cell wall formation in agaricomycete Pleurotus ostreatus

Hydrophobins are small-secreted proteins with both hydrophobic and hydrophilic regions, enabling the mycelium to break through the air-medium interface by reducing the medium surface tension. Over 20 putative hydrophobin-encoding genes have been predicted in the agaricomycete Pleurotus ostreatus. Three hydrophobin-encoding genes, vmh2, vmh3, and hydph16, were predominantly expressed in the vegetative mycelium. Despite these common properties, we have previously demonstrated the distinct functions of Vmh2 and Vmh3 in environmental stress resistance. In this study, we focused on hydph16 and found that Δhydph16 strains had sparser aerial mycelium than control strains. The cell wall thickness of Δhydph16 strains reduced by 40 % compared to that of control strains, but no significant differences were found in the relative chitin and glucan percentages or relative putative cell wall synthesis-related gene expression levels. Furthermore, unlike vmh2 and vmh3, hydph16 deletion did not change the hydrophobicity of the aerial mycelium. This study is the first to report that the lack of hydrophobin can lead to a significant change in aerial hyphae cell wall formation without altering the major cell wall polysaccharide composition. Additionally, this study revealed multiple roles for Hydph16, distinct from those of other highly expressed hydrophobins, Vmh2 and Vmh3. These results suggested that agaricomycetes, including P. ostreatus, have evolved to possess multiple hydrophobins with different functions.

Melatonin alleviates oxidative stress and inflammation of Leydig cells of Min pig through SIRT1 pathway

Inflammation responses and oxidative stress adversely affect testicular function, reducing fertility. Melatonin exhibits anti-inflammatory and antioxidant properties. However, the molecular mechanism by which melatonin alleviates inflammation and oxidative stress in Leydig cells of the Min pig testis remains unclear. To investigate this, primary Leydig cells were isolated from 7-day-ld Min pigs' testes and treated with LPS, H2O2 and melatonin, respectively. The results showed that co-treatment with melatonin and LPS significantly decreased the expression of TLR4, NF-κB, IL-6 and IL-1β compared to LPS group. Co-treatment with melatonin and H2O2 significantly mitigated reactive oxygen species and malondialdehyde levels. Melatonin also enhanced glutathione and superoxide dismutase levels and upregulated the mRNA expression levels of Nrf2, Keap1, HO-1 and NQO1. In the co-treatment group of melatonin, LPS, and SIRT1 inhibitor, the secretion of IL-6 and IL-1β and the mRNA expression levels of TLR4 and NF-κB were elevated significantly compared to the control group and the melatonin-LPS co-treatment group. In the combined treatment group of melatonin, H2O2 and a SIRT1 inhibitor, ROS levels increased significantly, while the expression of Nrf2, Keap1, HO-1 and NQO1 were decreased significantly compared to the control group and the melatonin-H2O2 co-treatment group. Furthermore, mRNA expression levels of testosterone synthesis-related genes StAR, CYP11A1, 3β-HSD, CYP17A1 and 17β-HSD significantly decreased following H2O2 treatment, which was alleviated by co-treatment with H2O2 and melatonin, but not by the addition of SIRT1 inhibitor. In conclusion, melatonin exhibits the capability to ameliorate inflammatory responses, oxidative stress and testosterone secretion in Leydig cells via the SIRT1 pathway.