STAR Protocols
·
2021
·
IF 0
Lidia Wrobel,Farah H. Siddiqi,David C. Rubinsztein
Summary We present a protocol for in vivo siRNA-mediated knockdown of a gene of interest in mouse liver using systemic delivery via intravenous injection. We describe a step-by-step protocol for delivery of siRNA particles, with tips on how to optimize dosage. We detail steps for feeding/starving cycles as well as for liver tissue isolation, followed by gene expression analysis, measured at the mRNA and protein levels. For complete information on the generation and use of this protocol, please refer to Wrobel et al. (2020) .
研究领域:
分子生物学
·
遗传学
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DOI
PubMed
Molecular & Cellular Toxicology
·
2017
·Vol. 13
·期 1
·59-64
·
IF 1.25
Chen Jiahong,Zhang Hang,Aniagu Stanley,Jiang Yan,Chen Tao
Trichloroethylene (TCE) had been shown to induce aberrant DNA methylation changes in mouse liver, which could be critical in the development of hepatocellular carcinoma. To understand the effect of methionine, a methyl donor, on TCE-induced mouse liver cancer, we exposed B6C3F1 mice to TCE in the presence or absence of methionine. We found that supplementation with methionine attenuated the TCE-induced hypo- and hyper-methylation status of Cdkn1a and Ihh in mouse liver, respectively. The TCE-induced mRNA expression changes of Cdkn1a, Ihh and Jun were also alleviated by methionine. However, methionine supplementation accelerated the TCE-induced mRNA over-expression of Mki67, and enhanced the TCE-induced mRNA down-regulation of Dnmt3a and Tet2. In summary, our results showed that methionine could attenuate at least part of the TCE-induced DNA methylation changes, but the additional gene expression changes induced by supplementary of methionine does not support the assumption that methionine could prevent TCE-induced liver carcinogenesis.
研究领域:
毒理学
·
分子生物学
·
癌症研究
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产品关联
DOI
高影响因子
Scientific Reports
·
2021
·Vol. 11
·期 1
·1-9
·
IF 4.38
Dong Wenjing,Song Erqun,Song Yang
The acute liver injury (ALI) and hepatic fibrosis caused by the co-treatment of lipopolysaccharide (LPS)/ d -galactosamine (D-GalN) have been extensively studied. However, whether LPS/D-GalN are genotoxic has been left unknown. In this study, male mice were divided into eight groups with eight animals in each group. For acute challenge of LPS/D-GalN, the mice in each group received a combination of LPS/D-GalN via intraperitoneal injection at the dose of 25 μg/kg/250 mg/kg, 25 μg/kg/500 mg/kg, or 50 μg/kg/500 mg/kg body weight. An additional group for chronic administration of test compounds was conducted by i.p. injection of LPS/D-GalN (10 μg/kg/100 mg/kg) every other day for 8 weeks. Saline solution (0.9%) and cyclophosphamide (CTX) (50 mg/kg body weight) given by i.p. injection was used as the negative and positive control, respectively. The results of single cell gel electrophoresis (SCGE) assay indicated that acute exposure of the mice to LPS/D-GalN caused severe DNA damage in hepatic cells, but not in the brain, sperm or bone marrow cells, which evidenced the genotoxicity of LPS/D-GalN administrated in combination. Interestingly, the chronic administration of LPS/D-GalN triggered significant genotoxic effects not only in hepatic but also in brain cells, with negative results in sperm and bone marrow cells. Histopathological examination in the liver and brain tissues revealed changes consistent with the SCGE results. The present study indicates genotoxic potential of LPS/D-GalN co-administered in mice, which may serve as an in vivo experimental model for relevant genotoxic study.
研究领域:
毒理学
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分子生物学
·
医学
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL PHARMACOLOGY
·
2023
·
IF 5.8
Luomin Lin,Yuwei Huang,Jinyi Wang,Xiaocao Guo,Fangjun Yu,Di He,Chuanbin Wu,Lianxia Guo,Baojian Wu
CYP2A5 , an enzyme responsible for metabolism of diverse drugs, displays circadian rhythms in its expression and activity. However, the underlying mechanisms are not fully established. Here we aimed to investigate a potential role of CRY1/2 (circadian clock modulators) in circadian regulation of hepatic CYP2A5. Regulatory effects of CRY1/2 on CYP2A5 were determined using Cry1 -null and Cry2 -null mice, and validated using AML-12, Hepa1-6 and HepG2 cells. CYP2A5 activities both in vivo and in vitro were assessed using coumarin 7-hydroxylation as a probe reaction. mRNA and protein levels were detected by qPCR and western blotting , respectively. Regulatory mechanism was studied using a combination of luciferase reporter assays, chromatin immunoprecipitation (ChIP) and co-immunoprecipitation (Co-IP). We found that ablation of Cry1 or Cry2 in mice reduced hepatic CYP2A5 expression (at both mRNA and protein levels) and blunted its diurnal rhythms. Consistently, these knockouts showed decreased CYP2A5 activity (characterised by coumarin 7-hydroxylation) and a loss of its time-dependency, as well as exacerbated coumarin-induced hepatotoxicity. Cell-based assays confirmed that CRY1/2 positively regulated CYP2A5 expression and rhythms. Based on combined luciferase reporter, ChIP and Co-IP assays, we unraveled that CRY1/2 interacted with E4BP4 protein to repress its inhibitory effect on Cyp2a5 transcription and expression . In conclusion, CRY1/2 regulate rhythmic CYP2A5 in mouse liver through repression of E4BP4. These findings advance our understanding of circadian regulation of drug metabolism and pharmacokinetics .
研究领域:
药理学
·
分子生物学
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时间生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Miao Zhang,Chenxu Wang,Lin Zhu,Hanshu Zhu,Wenhao Zheng,Weiying Lu,Yuge Niu,Yaqiong Zhang,Boyan Gao,Liangli Lucy Yu
As a part of a continuous 90 day subchronic toxicology study, integrated metabolomic and transcriptomic approaches were applied to assess the metabolic network changes in the livers of Kunming mice exposed to three typical thermally induced food toxicants, including oxidative derivatives of triacylglycerols (ox-TGs), aldehydes, and 3-monochloropropane-1,2-diol esters (3-MCPDE), as well as their mixtures. Results showed lipid metabolic dysregulation through impaired purine metabolism, PPAR signaling, and bile acid metabolism. Ox-TGs emerged as the most hazardous compound, altering over 10 genes/enzymes. 3-MCPDE exhibited gender-specific effects, significantly upregulating fatty acid metabolism and gluconeogenesis genes in males. Interestingly, toxicant mixtures attenuated the adverse metabolic effects caused by individual compounds, demonstrating complex regulatory mechanisms in fatty acid biosynthesis and oxidation. The metabolomic and transcriptomic analyses conducted in this study revealed that combined exposure to multiple toxicants generated during lipid thermal processing may induce more complex toxicity effects than the simple additive effects of individual toxicants. Certain antagonistic effects were observed when comparing individual toxicants to their mixtures, highlighting the need for further mechanistic verifications in this area.
研究领域:
毒理学
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生物化学
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分子生物学
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营养学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Yingxin Chen,Qianying Sheng,Yayun Hong,Minbo Lan
A hydrophilic nanocomposite was synthesized by an easy route to improve glycopeptides enrichment efficiency. This new composite, prepared with a method based on electrostatic interaction, was demonstrated to be efficient for immobilization of carrageenan on graphene oxide/poly(ethylenimine) support (denoted as GO–PEI–Carr). Carrageenan, which has a large number of hydroxyl groups and is fully negatively charged, is a new modified phase of hydrophilic materials in glycoproteomics. The introduction of carrageenan provided the composite not only a perfect surface charge but also a greater ability to enrich glycosylated peptides. Thirty-four glycopeptides from human serum immunoglobulin G (IgG) tryptic digests were obviously observed with greatly improved signal-to-noise (S/N) ratio. A good selectivity was still kept even when the molar ratio of IgG and bovine serum albumin (BSA) tryptic digest mixtures reached to 1:500. Meanwhile, 76 glycopeptides derived from 56 glycoproteins with 83 N-glycosylation sites were identified from human serum and 149 glycopeptides derived from 129 glycoproteins with 157 N-glycosylation sites were identified from mouse liver tissues, which showed the ability to enrich glycopeptides from complex biological samples. In addition, GO–PEI–Carr exhibited a unique repeatability and stability even after enrichment of glycopeptides for 20 times. It also performed a higher sensitivity (1 fmol/μL IgG), a better enrichment capacity (up to ∼300 mg/g), and an ideal enrichment recovery (90.8% and 109.5%) for glycopeptides enrichment, indicating a great potential for the application of glycoproteomic research.
研究领域:
材料科学
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分析化学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2024
·Vol. 14
·期 1
·1-14
·
IF 3.8
He Keting,Wei Dongfan,Liu Qiuhong,Liu Xiaoli,Zhou Diwenxin,Chen Shangci,Zhu Danhua,Xu Xiaowei
Acute liver injury (ALI) presents a challenging problem worldwide, prompting extensive research efforts. Cellular senescence has been found to be induced following ALI, and targeting cellular senescence has shown therapeutic potential. Therefore, understanding the expression of senescence-related genes in ALI can help to explore pathogenesis and treatment of this common disease. Carbon tetrachloride (CCl 4 ) is commonly used to study ALI. Although polymerase chain reaction (PCR) is a convenient and economical molecular biology technique widely used in basic medicine, research on selecting suitable reference genes to obtain objective and reproducible PCR data is scarce. Moreover, evidence supporting the choice of reference genes for experimental studies of CCl4-induced ALI and hepatic senescence in mice is limited. In this study, we obtained murine livers at four time points (0, 12, 24, and 48 h) following CCl4 treatment. We used five algorithms (geNorm, BestKeeper, NormFinder, delta Ct, and RefFinder) to rank 12 candidate genes in real-time reverse-transcription quantitative PCR (RT-qPCR) experiments. Focusing on cellular senescence in this model, we adopted four senescence-associated secretory phenotype (SASP) genes ( Il6 , Il1b , Ccl2 , and Ccl5 ) as target genes. Our results confirmed Gapdh and Tbp as suitable reference genes in murine CCl 4 -induced ALI models. Furthermore, we provide a table of published studies recommending reference genes for various liver disease models. This study provides a valuable reference for enhancing the reliability and reproducibility of ALI molecular findings.
研究领域:
分子生物学
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肝病学
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病理学
·
基因组学
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产品关联
DOI
PubMed
高影响因子
AMINO ACIDS
·
2023
·Vol. 55
·期 4
·499-507
·
IF 3.5
Qin Tiansheng,Huang Xiande,Zhang Qianqian,Chen Fan,Zhu Jiaojiao,Ding Yaoyao
During trauma and surgery, bleeding is a major concern. One of the crucial strategies for hemostasis is the use of biological hemostatic material. Herein, we reported an amino acid-based hydrogel FmocF-ADP hydrogel, which consisted of N-[(9H-fluoren-9-ylmethoxy) carbonyl]-3-phenyl-L-alanine (FmocF) and adenosine diphosphate (ADP) sodium solution. The hydrogel was created by FmocF self-assembling to nanofiber in ADP sodium solution and then cross-linking to hydrogel. FmocF-ADP hydrogel showed good in vitro coagulation activity as measured by whole blood clotting assays, platelet clotting assays, platelet activation assays, and platelet adhesion assays. Further, it was noted to reveal an exceptional in vivo hemostatic effect in a mouse liver bleeding model. Together with the previous report of the good biocompatibility and antimicrobial activity of FmocF hydrogel, our study would extend the biomedical application of FmocF hydrogel. In conclusion, the present study would provide a constructive strategy for the development of new antimicrobial and hemostatic materials or develop a potential hemostatic material.
研究领域:
生物医学工程
·
材料科学
·
止血
·
药剂学
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DOI
PubMed
高影响因子
Advanced Science
·
2022
·Vol. 9
·期 26
·2202505
·
IF 17.521
Guofeng Liu,Jinsong Wei,Xiaoyu Li,Meng Tian,Zhenxing Wang,Congcong Shen,Wan Sun,Chonghui Li,Xuewen Li,Enguang Lv,Shizheng Tian,Jihua Wang,Shicai Xu,Bing Zhao
Exploring the advanced techniques for protein detection facilitates cell fate investigation. However, it remains challenging to quantify and visualize the protein with one single probe. Here, a luminescent approach to detect hepatic cell fate marker albumin in vitro and living cell labeling with upconversion nanoparticles (UCNPs), which are conjugated with antibody (Ab) and rose bengal hexanoic acid (RBHA) is reported. To guarantee the detection quality and accuracy, an “OFF–ON” strategy is adopted: in the presence of albumin, the luminescence of nanoparticles remains suppressed owing to energy transfer to the quencher. Upon albumin binding to the antibody, the luminescence is recovered under near-infrared light. In various bio-samples, the UC NPs- A b- R BHA (UCAR) nanoprobe can sense albumin with a broad detection range (5–315 ng mL −1 ). When applied to liver ductal organoid culture medium, the UCAR can monitor hepatocyte differentiation in real time by sensing the secreted albumin. Further, UCAR enables live imaging of cellular albumin in cells, organoids, and tissues. In a CCl 4 -induced liver injury model, UCAR detects reduced albumin in liver tissue and serum. Thus, a biocompatible nanoprobe for both quantification and imaging of protein in complex biological environment with superior stability and high sensitivity is provided.
研究领域:
生物学
·
生物化学
·
纳米技术
·
医学成像
·
细胞生物学
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产品关联
DOI
PubMed
高影响因子
ACS Applied Polymer Materials
·
2024
·
IF 4.4
Xiaoyan Song,Siyuan Yu,Junxia Deng,Jia Liu,Jinfeng Xing
The ideal hemostatic dressing should not only achieve the effect of hemostasis but also maintain a suitable moist environment and sufficient physical and mechanical strength and avoid secondary injury to the wound. Here, a stretchable, adhesive, and antibacterial hydrogel (SAQH) for wound hemostasis is facilely prepared by photopolymerization under green LED irradiation, in which acrylamide (AAM) and methacryloylethylsulfobetaine (SBMA) are used as the skeleton and quaternized carboxymethyl chitosan (QCCS) is used as the procoagulant component. The introduction of the macromolecular cross-linking agent methacryloyl hyaluronic acid (HA-GMA) enhances biocompatibility and avoids the addition of small molecular toxic cross-linking agents. The introduction of inorganic nanoparticle hydroxyapatite (HAP) into the hydrogel system not only improves the mechanical properties of the polymer network but also releases calcium ions from HAP to achieve a good coagulation-promoting effect. SAQH hydrogels have excellent mechanical properties (the maximum stress can reach 237 kPa), good adaptability, sufficient tissue adhesion, and effective antibacterial properties. In the mouse liver injury model, hydrogels can adhere to the bleeding site to reduce blood loss, and they will not cause secondary bleeding when removed. This research develops a potential hemostatic hydrogel with broad applicable prospects.
研究领域:
生物材料
·
组织工程
·
伤口愈合
·
止血
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Pei-Fang Cai,Bing-De Zheng,Ya-Ling Xu,Bao-Xia Li,Zhi-Yu Liu,Ya-Yan Huang,Jing Ye,Mei-Tian Xiao
Collagen (COL) is the most widespread functional protein. Designing and developing dual-dynamic-bond cross-linked COL adhesive hydrogel sealants with multifunctional is highly advantageous for achieving a superior wound closure effect and hemostasis. In this study, we developed hybrid hydrogels consisting of fish-skin COL, oxidized sodium alginate (OSA), borax and polyvinyl alcohol (PVA) to enhance full-thickness wound healing. The hydrogels were furnished with first-rate self-healing capabilities through the dual-dynamic-bond cross-linking of dynamic Schiff base bonds (COL-OSA) and diol boric acid bonds (OSA-borax) with reversible breakage and re-formation. Moreover, the incorporation of PVA stimulated the formation of hydrogen bonds in the system, bolstering the stability of the hydrogel framework. The prepared hydrogel manifests self-healing, injectability, multifunctional adhesiveness and biodegradability. In vivo assessment of the hemostatic capacity of COSP 20 hydrogel was superior to gauze both in the mice liver injury model and mice tail amputation model. In addition, a full-thickness skin wound model in mice revealed that the COSP 20 hydrogel facilitated faster wound closure by accelerating reepithelialization, COL deposition and angiogenesis. These findings illustrate the potential of hybrid fish-skin COL-based hydrogels to enhance wound healing and promote rapid tissue repair, and provide new possibilities for the effective utilization of marine fishery resources.
研究领域:
生物材料
·
伤口愈合
·
止血
·
生物医学工程
·
组织工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）