Inventions
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2019
·Vol. 4
·期 4
·65
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IF 0
Fiorency Santoso,Bonifasius Putera Sampurna,Yu-Heng Lai,Sung-Tzu Liang,Erwei Hao,Jung-Ren Chen,Chung-Der Hsiao
This study aimed to develop a simple and cost-effective method to measure blood flow in zebrafish by using an image-based approach. Three days post fertilization (dpf) zebrafish embryos were mounted with methylcellulose and subjected to video recording for tracking blood flow under an inverted microscope equipped with a high-speed CCD camera. In addition, Hoffman lens was used to enhance the blood cell contrast. The red blood cell movement was tracked by using the TrackMate plug-in in the ImageJ image processing program. Moreover, Stack Difference and Time Series Analyzer plug-in were used to detect dynamic pixel changes over time to calculate the blood flow rate. In addition to blood flow velocity and heart rate, the effect of drug treatments on other cardiovascular function parameters, such as stroke volume and cardiac output remains to be explored. Therefore, by using this method, the potential side effects on the cardiovascular performance of ethyl 3-aminobenzoate methanesulfonate (MS222) and 3-isobutyl-1-methylxanthine (IBMX) were evaluated. MS222 is a common anesthetic, while IBMX is a naturally occurring methylxanthine. Compared to normal embryos, MS222- and IBMX-treated embryos had a reduced blood flow velocity by approximately 72% and 58%, respectively. This study showed that MS222 significantly decreased the heart rate, whereas IBMX increased the heart rate. Moreover, it also demonstrated that MS222 treatment reduced 50% of the stroke volume and cardiac output. While IBMX decreased the stroke volume only. The results are in line with previous studies that used expensive instruments and complicated software analysis to assess cardiovascular function. In conclusion, a simple and low-cost method can be used to study blood flow in zebrafish embryos for compound screening. Furthermore, it could provide a precise measurement of clinically relevant cardiac functions, specifically heart rate, stroke volume, and cardiac output.Keywords:blood flow tracking;ImageJ;zebrafish;IBMX;MS222;cardiovascular function assessment
研究领域:
发育生物学
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毒理学
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MICROCHIMICA ACTA
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2018
·Vol. 185
·期 4
·1-7
·
IF 5.705
Zhang Kaina,Zeng Ke,Shen Congcong,Tian Shiyu,Yang Minghui
The authors describe a fluorometric method for the determination of the activity and inhibition of protein kinase A (PKA). In the presence of ATP, PKA catalyzes the transfer of phosphate groups from ATP to a peptide, and the generated phosphorylated peptide quenches the fluorescence (measured at excitation/emission peaks of 340/440 nm) of the hydroxyapatite nanoparticles (HAP-NPs). A linear logarithmic relationship of PKA concentrations with fluorescence intensity in the range from 1 to 50 U·L −1 was obtained, and the lower limit of detection (LOD) is 0.5 U·L −1 . This is much lower than LODs reported in the literature. The PKA inhibitor H-89 was studied, and the inhibition plot has a sigmoidal shape with a half-maximal inhibitory concentration of around 750 nM of H-89. At a 4.5 nM level of H-89, fluorescence of HAP-NPs fell to levels of no PKA controls, demonstrating that the assay is a viable tool to screen for kinase inhibitors. An assay with Hela cell lysates in combination with forskolin (an activator of adenylyl cyclase) and IBMX (a phosphodiesterase inhibitor used to activate the cellular activity of PKA) resulted in decreased fluorescence of HAP-NPs. This suggests that the assay can be applied for testing in vitro cell kinase activity. In our perception, this method will enable high-throughput screening for kinase-related drugs and fluorometric enzymatic detection in various areas.
Fluorescence assay based on hydroxyapatite nanoparticles (HAP) fluorescence quenching was developed for analysis of the activity and inhibition of protein kinase A (PKA).
研究领域:
生物化学
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分子生物学
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药理学
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分析化学
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PubMed
高影响因子
Endocrine Metabolic & Immune Disorders-Drug Targets
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2025
·Vol. 26
·期 1
·E18715303382831
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IF 2
Yuting Chen,Jie Min,Xiali Yu,Haixiang Ni,Yamei Jin
IntroductionAn increase in the intraorbital adipose tissue is the main pathological feature of thyroid-associated ophthalmopathy (TAO). IGF-1R activates PI3K/AKT signaling and accelerates adipogenesis. Pingmu decoction has been demonstrated to promote orbital adipocyte apoptosis; however, less is reported regarding the action mechanism of Danshenol A (DA), a single active ingredient of Salvia miltiorrhiza (Danshen). Accordingly, this study aimed to investigate the role and association of DA and IGF-1R in the proliferation and lipid accumulation of orbital adipocytes.MethodsPrimary human orbital preadipocytes were chosen and authenticated using immunofluorescence. Cells were treated with the IGF-1R agonist ginsenoside Rg5, IGF-1R overexpression plasmid, dexamethasone (Dex), and/or DA, after which cell proliferation and differentiation were assessed by cell counting kit-8 (CCK-8), oil red O staining, real-time quantitative polymerase chain reaction, and Western blot assays.ResultsOrbital preadipocytes showed positive expression of Pref-1. Treatment with IGF-1R agonist, as well as Dex, promoted orbital adipocyte viability and lipid accumulation, and increased the expression of adiponectin and leptin. It was observed that the overexpression of IGF-1R boosted PI3K/AKT activation and elevated PPARγ and C/EBPα expressions. Importantly, DA reversed the effects of IGF-1R on cell viability, lipid accumulation, and the PI3K/AKT signaling pathway.DiscussionThis study, for the first time, revealed the molecular mechanism by which DA regulates orbital fat metabolism through targeted inhibition of the IGF-1R/PI3K/AKT signaling axis. Notably, IGF-1R overexpression partially counteracted the inhibitory effect of DA, suggesting that this component has multi-target regulatory characteristics.ConclusionThis study not only reveals a new mechanism by which DA treats TAO but also provides theoretical support for the treatment of adipose metabolism.
研究领域:
内分泌学
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眼科学
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分子生物学
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信号转导
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脂肪代谢
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中药药理学
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PubMed
高影响因子
Nature Communications
·
2026
·
IF 18.1
Mao Congyang,Jin Wanyu,Dou Limeng,Guo Tong,Huang Jin,Wang Yi,Liu Xiangmei,Wu Shuilin,Qiao Wei,Xiang Yiming,Zhu Yizhou,Wu Jun,Yeung Kelvin Wai Kwok
The pandemic-scale progression of type 2 diabetes mellitus (T2DM) necessitates innovative interventions targeting the pathogenic triad of insulin resistance, dysregulation of lipid metabolism, and gut microbiome dysbiosis. Here, we report a synthetically bioengineered probiotic consortium (REcN-F/Ca) developed through directed metabolic adaptations of Escherichia coli Nissle 1917 (EcN) under iterative hydrogen peroxide selection, subsequently functionalized with fructooligosaccharide-calcium carbonate composites. REcN-F/Ca exhibits enhanced reactive oxygen species tolerance through upregulated antioxidant enzymes and hydrogen sulfide-mediated redox balancing, alongside improved gastrointestinal survivability. In high-fat diet-induced obese male mice, REcN-F/Ca restores gut microbiota diversity, enriches butyrogenic taxa (Lachnospiraceae and Blautia ), and rescues short-chain fatty acids depletion. Transcriptomic profiling reveals PPAR signaling activation, driving lipid metabolism and suppressing adipose inflammation. These effects translate to systemic metabolic improvements with attenuated weight gain (−25.4%), restored glucose homeostasis, and reduced insulin resistance (HOMA-IR: −73.2%) in the obesity and T2DM murine model. Our findings establish REcN-F/Ca as a synthetically engineered probiotic that simultaneously corrects intestinal ecological perturbations and reverses host metabolic dysfunction, proposing a paradigm for metabolic syndrome management.
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PHYTOTHERAPY RESEARCH
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2026
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IF 8.1
Jing Li,Yang-Chi-Dung Lin,Hsi-Yuan Huang,Jihang Chen,Huali Zuo,Yun Tang,Hsien-Da Huang
Obesity is a major risk factor for type 2 diabetes. Some traditional Chinese medicine (TCM) compounds can improve obesity by increasing energy consumption. This study aimed to identify and verify TCM compounds that promote adipose thermogenesis to alleviate obesity via transcriptomic analysis, molecular docking, network pharmacology, and Connectivity Map (CMap) analysis. Thirty-six TCMs related to adipose tissue thermogenesis were first collected to generate transcriptomic data. Then a ranking method based on integrated transcriptomic data was used to select emodin from the TCM Da Huang for network pharmacology to explore its potential mechanisms and further experimental validation. CMap analysis of thermogenesis signature genes from ProFAT and GEO datasets identified triptolide as another compound for improving obesity through enhanced thermogenesis. In vitro experiments with 3T3-L1 cells and in vivo experiments with zebrafish were conducted. The results showed that both emodin and triptolide could upregulate the NAD + /NADH ratio, increase AMPK and SIRT1 expression, suppress lipid accumulation, and promote lipolysis, as verified by both in vitro and in vivo experiments. They also upregulated thermogenesis-related genes (e.g., UCP1, PGC1α, and PRDM16), lipolysis-related genes (e.g., PKA, ATGL, and HSL), mitochondrial biogenesis-related genes (e.g., NRF1, NRF2, and TFAM), and β-oxidation-related genes (e.g., CPT1α and CPT1β). Meanwhile, they downregulated lipogenesis-related genes such as FASN, SREBP1, and ACC. These results provide a basis for understanding the effects of emodin and triptolide on obesity. The findings highlight the effectiveness of combined screening approaches in exploring compounds with beneficial metabolic effects.
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高影响因子
FREE RADICAL BIOLOGY AND MEDICINE
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2025
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IF 8.2
Longyan Yang,Ruili Yin,Ziyu Shan,Anqi Feng,Kun Li,Yan Wang,Ke Yu,Yongsong Xu,Lin Mao,Jianan Lang,Baoyu Zhang,Dong Zhao
Aims Overweight and obesity are closely associated with hyperuricemia (HUA). However, the effect of obesity-induced adipokine on the level of serum uric acid (UA) has not been fully elucidated. This study aimed to determine the role of adipokine in the pathogenesis of HUA. Methods Omental adipose tissues from HUA patients with obesity were collected for data- independent acquisition (DIA) proteomics analysis, and the secreted protein angiopoietin-like protein (ANGPTL2) was identified and further validated via Western blot and ELISA. Angptl2 knockout (ko) and adipocyte-specific ANGPTL2 overexpression mice were generated to establish HUA models. HK2 cells and primary renal tubule epithelial cells (RTECs) were applied to induce HUA cell model and treated with recombinant human ANGPTL2 or supernatants derived from C3H10 cells differentiated into adipocytes. Result ANGPTL2 levels were significantly elevated in the omental adipose tissue from HUA with obesity patients compared to control participants. Circulating ANGPTL2 levels were also increased, and positively correlated with serum UA levels, especially in male participants. Plasma UA levels were significantly decreased in Angptl2 ko mice, coinciding with up-regulated expression of ATP binding cassette subfamily G member 2 (ABCG2) and down-regulated glucose transporter 9 (GLUT9) in renal tissue. However, plasma UA levels were significantly increased, accompanied by decreased ABCG2 expression and enhanced GLUT9 expression after adipocyte-specific ANGPTL2 overexpression. Furthermore, adipocyte- derived ANGPTL2 increased UA level in cell supernatants through the inhibition of protein kinase B (AKT)/ABCG2 signaling in RTECs and HK2 cells. Conclusion Adipokine ANGPTL2 levels were significantly elevated in HUA patients with obesity. Circulating ANGPTL2 concentrations were independently associated with serum UA levels, particularly in obese males. Adipocyte-secreted ANGPTL2 promoted the level of UA through AKT/ABCG2 signaling in renal tubular cells. These findings establish ANGPTL2 as a critical adipokine linking obesity to hyperuricemia and suggest its potential as a novel therapeutic target for HUA.
研究领域:
内分泌学
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代谢研究
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分子生物学
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PubMed
高影响因子
Nature Communications
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2025
·Vol. 16
·期 1
·1-12
·
IF 15.7
Tian Ran,Zhang Tong,Dong Hao,Seim Inge,Yang Guang
Blubber is a hydrostatic pressure-resistant and insulating vascularized layer of subcutaneous adipose tissue beneath the skin found in cetaceans, pinnipeds, and sirenians. Little is known about the molecular genetics of blubber formation, and it is unclear when the blubber observed today began to develop during the evolution of the marine mammal groups. Here we screen for genes lost in cetaceans and find that ADRB3 (β 3 -adrenergic receptor), a gene essential for adipose tissue lipolysis and thermogenesis, acquired inactivating mutations in crown cetaceans 20 million years after its downstream effector uncoupling protein 1 ( UCP1 ) gained partial loss-of-function mutations in semi-aquatic stem cetaceans. Both in vivo and in vitro experiments indicate that cetacean ADRB3 is not functional, suggesting that crown cetaceans abolished cold-induced non-shivering thermogenesis to favor lipid storage for thermal insulation. Our study supports that cetacean blubber evolved stepwise along their transition to an aquatic environment and implies that ADRB3 loss was critical for the radiation and dispersal of toothed and baleen whales.
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高影响因子
Diabetes & Metabolism Journal
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2025
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IF 8.5
Lei Zhou,Yuwen Jiao,Jiaming Xue,Xiaoqiang Zhan,Dongmei Wang,Liming Tang
Background: To investigate the biological functions of the transcription factor LRR binding FLII interacting protein 1 (LRRFIP1) in white adipocyte differentiation (WAD) and elucidate the underlying molecular regulatory mechanisms involved. Methods: Consensus clustering, differential gene expression screening, and intersection analysis were used to identify transcription factors involved in WAD. Adipogenic differentiation experiments were conducted using C3H10T1/2 cells, and a diet-induced obesity model in C57BL/6J mice was established to investigate the function of LRRFIP1 in WAD in vitro and in vivo. Molecular mechanisms were examined through quantitative real-time polymerase chain reaction, Western blotting, luciferase assays, and chromatin immunoprecipitation. Results: Bioinformatics analyses identified LRRFIP1 as a transcription factor associated with WAD. LRRFIP1 expression was downregulated in white adipose tissues from obese patients and in mature white adipocytes. Silencing LRRFIP1 significantly inhibited WAD in C3H10T1/2 cells and reduced differentiation biomarker expression; in contrast, overexpressing LRRFIP1 had the opposite effects. Mechanistically, LRRFIP1 bound to the E2F transcription factor 6 (E2F6) promoter to suppress E2F6 transcription, thereby downregulating a key differentiation regulator, CCAAT enhancer binding protein alpha (C/EBPα). Furthermore, in a diet-induced obesity model, LRRFIP1 could regulate the differentiation and maturation of inguinal white adipose tissue. Conclusion: Our findings reveal that LRRFIP1 plays a crucial inhibitory role in WAD by negatively regulating the E2F6/C/EBPα axis. This discovery not only enriches our understanding of the molecular networks governing WAD but also holds great promise for creating targeted therapies for obesity and associated metabolic conditions.
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高影响因子
BIOLOGICAL & PHARMACEUTICAL BULLETIN
·
2020
·Vol. 43
·期 7
·1027-1034
·
IF 1.863
Mingzhi Luo,Peili Yu,Kai Ni,Yang Jin,Lei Liu,Jingjing Li,Yan Pan,Linhong Deng
Excessive contraction of airway smooth muscle cells (ASMCs) is a hallmark feature of asthma. Intriguing, the activation of bitter taste receptor (TAS2R) in ASMCs can relax ASMCs. However, there is a lack of potent TAS2R agonists that can be used in asthma therapies since those tested agonists cannot relax ASMCs at the dose below a few hundred micromolar. Considering that sanguinarine (SA) is a bitter substance often used in small doses for the treatment of asthma in folk medicine, the present study was to determine the rapid relaxation effect of SA on ASMCs and to reveal the underlying mechanisms associated with TAS2R signaling. Here, cell stiffness, traction force, calcium signaling, cAMP levels, and the mRNA expression were evaluated by using optical magnetic twisting cytometry, traction force microscopy, Fluo-4/AM labeling, enzyme-linked immunosorbent assay (ELISA), and quantitative (q)RT-PCR, respectively. We found that 0.5 µM SA immediately decreased cell stiffness and traction force, which is comparable with the effect of 5 µM isoproterenol. In addition, 0.5 µM SA immediately increased intracellular free calcium concentration ([Ca2+]i) and decreased the mRNA expression of contractile proteins such as calponin and α-smooth muscle actin after the treatment for 24 h. Furthermore, SA-mediated decrease in cell stiffness/traction force and increase in [Ca2+]i were significantly blunted by inhibiting the TAS2Rs signaling. These findings establish the rapid relaxation effect of SA at low concentration (<1 µM) on cultured ASMCs depending on TAS2R signaling, indicating that SA might be developed as a useful bronchodilator in asthma therapy.
研究领域:
药理学
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呼吸医学
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细胞生物学
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PubMed
高影响因子
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS
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2025
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IF 3.3
Qinghua Fu,Peng Wang,Weilin Li,Zhenhua Cai,Shiji Zhao,Weidong Ling,Mingxun Li,Xiaochuan Tang,Ziyi Song
Obesity poses a significant risk for metabolic disorders, such as insulin resistance and metabolic-associated fatty liver disease (MAFLD), yet effective treatments remain limited. Cell Death-Inducing DNA Fragmentation Factor-α-Like Effector C (CIDEC), a lipid droplet membrane protein, facilitates lipid droplet fusion and is crucial for adipose tissue expansion, making it a key target for obesity and related metabolic diseases. However, previous research revealed that complete genetic deletion of Cidec in adipose tissues, while reducing fat accumulation, induced severe insulin resistance in high-fat diet (HFD)-fed mice, potentially due to ectopic fat storage in the liver. Given that complete knockout is an extreme approach, partial inhibition holds greater clinical relevance. Therefore, this study aimed to investigate the effects of partial inhibition of CIDEC in adipose tissues on fat accumulation and insulin sensitivity in mice. Using the Cre-LoxP system, we generated adipose Cidec haploinsufficient mice. Under a standard diet, these mice exhibited normal body weight, fat accumulation, and insulin sensitivity. Notably, under HFD conditions, mice with partial Cidec deficiency showed reduced fat accumulation in adipose tissues while hepatic fat accumulation remained unchanged, accompanied by improved insulin sensitivity and increased energy expenditure. Mechanistically, we found partial Cidec deficiency activated thermogenic program in adipocytes in vivo and in vitro through the ATGL-PPARα pathway. In conclusion, adipose CIDEC partial inhibition attenuates HFD-induced obesity and insulin resistance by enhancing ATGL-PPARα-mediated energy expenditure, establishing this approach as a promising therapeutic strategy for obesity and related metabolic diseases.
研究领域:
生物学
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医学
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代谢疾病
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF OBESITY
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2024
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IF 4.2
Yan Zheng,Luo Jia,Wang Ying,Yang Jie,Su Mingli,Jiang Lei,Yang Julin,Dai Manyun,Liu Aiming
Background As the second most risky environmental pollution, noise imposes threats to human health. Exposure to high-intensity noise causes hearing impairment, psychotic disorders, endocrine modifications. The relationship among low-intensity noise, obesity and lipid-regulating nuclear factor PPARα is not yet clear. Methods In this study, male wild-type (WT) and Pparα -null (KO) mice on a high-fat diet (HFD) were exposed to 75 dB noise for 12 weeks to explore the effect of low-intensity noise on obesity development and the role of PPARα. 3T3-L1 cells were treated with dexamethasone (DEX) and sodium oleate (OA) to verify the down-stream effect of hypothalamic-pituitary-adrenal (HPA) axis activation on the adipose tissues. Results The average body weight gain (BWG) of WT mice on HFD exposed to noise was inhibited, which was not observed in KO mice. The mass and adipocyte size of adipose tissues accounted for the above difference of BWG tendency. In WT mice on HFD, the adrenocorticotropic hormone level was increased by the noise challenge. The aggravation of fatty liver by noise exposure occurred in both mouse lines, and the transport of hepatic redundant lipid to adipose tissues were similar. The lipid metabolism in adipose tissue driven by HPA axis accorded with the BWG inhibition in vivo, validated in 3T3-L1 adipogenic stem cells. Conclusion Chronic exposure to low-intensity noise aggravated fatty liver in both WT and KO mice. BWG inhibition was observed only in WT mice, which covered up the aggravation of fatty liver by noise exposure. PPARα mediates the activation of HPA axis by noise exposure in mice on HFD. Elevated adrenocorticotropic hormone (ACTH) promoted lipid metabolism in adipocytes, which contributed to the disassociation of BWG and fatty liver development in male WT mice. Summary of PPARα suppresses noise-induced body weight gain in mice on high-fat-diet. Chronic exposure to low-intensity noise exposure inhibited BWG by PPARα-dependent activation of the HPA axis.
研究领域:
环境健康
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内分泌学
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营养学
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分子生物学
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PubMed
高影响因子
Food Chemistry-X
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2024
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IF 6.5
Peng Wang,Haifan Wang,Yang Xiao,Jialiang Zou,Hongping Chen,Lin Chen,Fu Wang,Yuan Hu,Youping Liu
In this study, comprehensive and systematic nontargeted metabolomics analysis was performed with the metabolites of Zangju peel ( Citrus reticulata cv. Manau Gan, CRZP, which has been cultivated for over 400 years in Derong County, China.) at four different mature stages. A total of 1878 metabolites were identified, among which flavonoids were the most abundant (62.04 %), and identified 62 key differential metabolites significantly affected by maturity. Based on biological activity measurements, CRZP showed better antioxidant activity, lipase inhibition ability, inhibition of adipogenic differentiation in 3TT-L1 cells and promotion of lipid metabolism, with the biological activity of CRZP at different maturity stages being associated with key differential metabolite. Thus, CRZP is natural antioxidants and possess anti-obesity potential, and industrial production needs to consider the Maturity stage of its collection.
研究领域:
植物科学
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代谢组学
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药理学
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食品科学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）