BIOLOGICAL & PHARMACEUTICAL BULLETIN
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2020
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IF 1.863
Yuanguo Xiong,Hao Lu,Hanlin Xu
Hepatic fibrosis (HF) is a common disease, with currently no available treatment. Galangin, a natural flavonoid extracted from Alpinia officinaruim Hance, has multiple effects demonstrated in previous studies. The aim of the present study was to explore the anti-fibrogenic effect of galangin in vitro, and research its potential molecular mechanisms. LX-2 cells were chosen as an in vitro HF model, and were treated with galangin in different concentrations. Cell viability was analyzed using Cell Counting Kit-8 (CCK-8) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell apoptosis was measured using flow cytometry, and the anti-fibrogenic effect of galangin was determined using quantitative reverse transcription polymerase chain reaction (RT-qPCR), immunofluorescence, and western blotting. The results show that the proliferation of LX-2 cells was efficiently inhibited by galangin, and apoptosis was induced in a dose-dependent manner. Both the mRNA and protein expression of α-SMA and collagen I were markedly downregulated. Galangin also inhibited the PI3K/Akt and Wnt/β-catenin signaling pathways and increased the Bax/Bcl-2 ratio. The results of this study suggest that galangin has an anti-fibrogenic effect and may represent a promising agent in the treatment of hepatic fibrosis.
研究领域:
药理学
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细胞生物学
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分子生物学
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肝病学
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产品关联
DOI
PubMed
高影响因子
DRUG AND CHEMICAL TOXICOLOGY
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2025
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IF 1.9
Aicun Tang,Luying Wei,Jiayi Gao,Zheng Zhang,Bingyu Xia,Xin Li,Qiuyu Lu
To analyze tadehaginoside (TA) for its effect on liver fibrosis and the associated mechanism. Mice with CCl4-mediated liver fibrosis were administered TA treatment for 6 weeks. Histopathological alterations in liver tissues were evaluated, and serum markers of hepatic injury, as well as the hepatic profibrotic gene and protein levels were detected. Transforming growth factor-β1 (TGF-β1) was adopted for inducing LX-2 cells. Following a 24-h TA intervention, we assessed pro-fibrotic gene and protein activation, proliferation and levels of TGF-β1-activated LX-2 cells. miR-155 and FOXO3 expressions were quantified in mice liver tissues and cells. The targeting relation of miR-155 with FOXO3 was verified. After regulating miR-155 and FOXO3 expression, the function of TA in cell activation, proliferation, and collagen accumulation was evaluated. The results indicated that TA treatment markedly alleviated CCl4-mediated mouse liver injury, inhibited collagen fiber deposition, and downregulated the hepatic pro-fibrotic gene and protein levels. Additionally, TA treatment suppressed TGF-β1-induced cell activation and proliferation, thus significantly reducing pro-fibrotic marker levels within cells. Furthermore, treatment with TA down-regulated miR-155 and up-regulated FOXO3 within CCl4-induced liver tissues as well as TGF-β1-induced cells. However, miR-155 overexpression or FOXO3 silencing partially mitigated the inhibition of TA against TGF-β1-mediated activation, proliferation, and fibrosis in cells. In conclusion, TA inhibits the activation of hepatic stellate cell (HSC) via miR-155/FOXO3 axis, thereby exerting its anti-hepatic fibrosis effect.
研究领域:
药理学
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分子生物学
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肝脏病学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
DIGESTIVE AND LIVER DISEASE
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2021
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IF 4.088
Si-Biao Su,Lin Tao,Xiao-Le Liang,Wen Chen
Background Liver fibrosis is a common disease that can lead to hepatic failure. Aims Our aims were to reveal the role of GAS5 in the regulation of liver fibrosis. Methods LX-2 human hepatic satellite cells (HSCs) were cultured and activated using TGF-β1 treatment. A CCK-8 assay was performed to assess cell viability. A luciferase assay was employed to monitor the interactions between miR-433–3p and GAS5 or toll-like receptor 10 (TLR10). Western blotting and real-time quantitative PCR (RT-qPCR) were applied to detect the expression levels of α-SMA, Col. I, PCNA-, MMP2-, MMP9-, TLR10-, and NF-κB-related molecules at the protein and RNA levels. Results GAS5 and TLR10 were decreased while miR-433–3p was upregulated in TGF-β1-activated LX-2 cells. Upregulation of GAS5 or downregulation of miR-433–3p suppressed HSC activation, and luciferase assays indicated that miR-433–3p binds with GAS5 and the 3′-UTR of TLR10. MiR-433–3p upregulation and TLR10 downregulation rescued the impacts of GAS5 overexpression or miR-433–3p knockdown on LX-2 cells. Upregulation of GAS5 also suppressed the phosphorylation of NF-κB through the miR-433–3p/TLR10 axis. Conclusion LncRNA GAS5 exerts an inhibitory effect on HSC activation by suppressing NF-κB signalling through regulation of the miR-433–3p/TLR10 axis.
研究领域:
分子生物学
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细胞生物学
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医学遗传学
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DOI
PubMed
高影响因子
Acta Biomaterialia
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2023
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IF 9.7
Ke Zhou,Rongjian Ding,Xiwang Tao,Yuwen Cui,Jiquan Yang,Hongli Mao,Zhongwei Gu
ABSTRACT Despite 3D bioprinting having emerged as an advanced method for fabricating complex in vitro models, developing suitable bioinks that fulfill the opposing requirements for the biofabrication window still remains challenging. Although naturally derived hydrogels can better mimic the extracellular matrix (ECM) of numerous tissues, their weak mechanical properties usually result in architecturally simple shapes and patchy functions of in vitro models. Here, this limitation is addressed by a peptide-dendrimer-reinforced bioink (HC-PDN) which contained the peptide-dendrimer branched PEG with end-grafted norbornene (PDN) and the cysteamine-modified HA (HC). The extensive introduction of ethylene end-groups facilitates the grafting of sufficient moieties and enhances thiol-ene-induced crosslinking, making HC-PDN exhibits improved mechanical and rheological properties, as well as a significant reduction in reactive oxygen species (ROS) accumulation than that of methacrylated hyaluronic acid (HAMA). In addition, HC-PDN can be applied for the bioprinting of numerous complex structures with superior shape fidelity and soft matrix microenvironment. A heterogeneous and biomimetic hepatic tissue is concretely constructed in this work. The HepG2-C3As, LX-2s, and EA.hy.926s utilized with HC-PDN and assisted GelMA bioinks closely resemble the parenchymal and non-parenchymal counterparts of the native liver. The bioprinted models show the endothelium barrier function, hepatic functions, as well as increased activity of drug-metabolizing enzymes, which are essential functions of liver tissue in vivo . All these properties make HC-PDN a promising bioink to open numerous opportunities for in vitro model biofabrication. Statement of significance In this manuscript, we introduced a peptide dendrimer system, which belongs to the family of hyperbranched 3D nanosized macromolecules that exhibit high molecular structure regularity and various biological advantages. Specifically, norbornene-modified peptide dendrimer was grafted onto PEG, and hyaluronic acid (HA) was selected as a base material for bioink formulation because it is a component of the ECM. Peptide dendrimers confer the following advantages to bioinks: a) Geometric symmetry can facilitate construction of bioinks with homogeneous networks; b) abundant surface functional groups allow for abundant crosslinking points; c) the biological origin can promote biocompatibility. This study shows conceptualization to application of a peptide-dendrimer bioink to extend the Biofabrication Window of natural bioinks and will expand use of 3D bioprinting of in vitro models.
研究领域:
生物医学工程
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组织工程
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生物材料科学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Cell and Developmental Biology
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2022
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IF 6.081
Heming Ma,Xiaomei Wang,Xu Liu,Chang Wang,Xiuzhu Gao,Junqi Niu
Liver fibrosis (LF) is a major disease that threatens human health. Hepatic stellate cells (HSCs) contribute directly to LF via extracellular matrix (ECM) secretion. Moreover, RXRα is an important nuclear receptor that plays a key regulatory role in HSC activation. Meanwhile, microRNAs (miRNAs) have been identified as significant regulators of LF development. In particular, miR-654-5p is involved in cellular migration and proliferation, and via bioinformatics analysis, has been identified as a potential factor that targets RXRα in humans and in mice. However, the precise relationship between miR-654-5p and RXRα in the context of LF, remains unknown and is the primary focus of the current study. To establish in vitro activated cell model human primary HSCs were cultured in vitro and LX-2 cells were stimulated with recombinant human TGF-β1. mRNA and protein levels of RXRα, miR-654-5p and fibrogenic genes were compared in quiescent and activated HSCs. Moreover, after transfected with miR-654-5p mimics, the expression changes of above related genes in LX-2 cells were estimated. Meanwhile, cell proliferation and apoptosis were detected in miR-654-5p overexpressed LX-2 cells. Simultaneously, the targeted binding between miR-654-5p and RXRα was verified in LX-2 cells. Carbon tetrachloride (CCl 4 )-induced mouse model with liver fibrosis was use to research the role of the miR-654-5p in vitro . Our results show that miR-654-5p expression levels increased in activated human HSCs and TGFβ-treated LX-2 cells. Moreover, miR-654-5p mimics markedly promoted LX-2 cell proliferation while inhibiting their apoptosis. Accordingly, the expression levels of RXRα are decreased in activated HSCs and LX-2 cells. Additionally, dual-luciferase reporter assay results reveal direct targeting of RXRα by miR-654-5p. Similarly, in vivo miR-654-5p overexpression aggravates LF in mice that are intraperitoneally injected with CCl 4 . Taken together, our findings elucidated a novel molecular mechanism with potential use for treatment of LF.
研究领域:
分子生物学
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肝病学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
LIVER INTERNATIONAL
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2022
·Vol. 43
·期 2
·500-512
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IF 8.754
Mengli Yu,Xinjue He,Xin Song,Jianguo Gao,Jiaqi Pan,Tianyu Zhou,Qinqiu Wang,Wei Zhu,Huijian Ma,Hang Zeng,Chengfu Xu,Chaohui Yu
Background Biglycan (BGN) is a small leucine-rich proteoglycan that participates in the production of excess extracellular matrix (ECM) and is related to fibrosis in many organs. However, the role of BGN in liver fibrosis remains poorly understood. This study aimed to investigate the role and mechanism of BGN in liver fibrosis. Methods Human liver samples, Bgn −/0 (BGN KO) mice and a human LX-2 hepatic stellate cells (HSCs) model were applied for the study of experimental fibrosis. GEO data and single-cell RNA-seq data of human liver tissue were analysed as a bioinformatic approach. Coimmunoprecipitation, immunofluorescence staining, western blotting and qRT–PCR were conducted to identify the regulatory effects of BGN on heat shock protein 47 (HSP47) expression and liver fibrosis. Results We observed that hepatic BGN expression was significantly increased in patients with fibrosis and in a mouse model of liver fibrosis. Genetic deletion of BGN disrupted TGF-β1 pathway signalling and alleviated liver fibrosis in mice administered carbon tetrachloride (CCl 4 ). siRNA-mediated knockdown of BGN significantly reduced TGF-β1-induced ECM deposition and fibroblastic activation in LX-2 cells. Mechanistically, BGN directly interacted with and positively regulated the collagen synthesis chaperon protein HSP47. Rescue experiments showed that BGN promoted hepatic fibrosis by regulating ECM deposition and HSC activation by positively regulating HSP47. Conclusion Our data indicate that BGN promotes hepatic fibrosis by regulating ECM deposition and HSC activation through an HSP47-dependent mechanism. BGN may be a new biomarker of hepatic fibrosis and a novel target for disease prevention and treatment.
研究领域:
分子生物学
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肝病学
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细胞生物学
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DOI
PubMed
高影响因子
Redox Biology
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2021
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IF 11.799
Chen Jin,Pengfei Wu,Yushun Yang,Zhenxiang He,Hailiang Zhu,Zhen Li
Realizing the early diagnosis of acute liver injury has become a hotspot in recent years. Since the current indexes are not specific, developing novel probes with new recognition group remains a necessity. In this work, we developed a novel fluorescent probe, NNP , for the detection of ONOO − in acute liver injury (ALI). The in vitr o evaluation of NNP indicated the advantages including high sensitivity (LOD = 0.13 μ M), rapid response (<25 s), naked-eye (red to yellow), and stability under various pH (4–10) and time (>48 h) conditions. Accordingly, NNP achieved the dose-dependent detection of ONOO − in human hepatic stellate LX-2 cells. Further, in the ALI model mice, NNP could monitor the ONOO − level in the occurrence of ALI in situ (in the liver region) with a steady performance. Subsequent immunohistochemical and imaging studies confirmed that NNP could achieve the detection of endogenous ONOO − in ALI liver tissues. This work introduced a practical implement for the ONOO − detection in ALI as well as a referable example for the establishment of molecular indexes in pre-clinical diagnosis.
研究领域:
生物化学
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分子生物学
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医学诊断
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药理学
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DOI
PubMed
高影响因子
MOLECULAR AND CELLULAR ENDOCRINOLOGY
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2025
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IF 3.6
Yujing Wang,Yuanxin Xu,Yilin Liu,Jindan Lin,Xincai Hong,Shuang Yan
Metabolic dysfunction-associated steatotic liver disease (MASLD) is a global disease with a broad spectrum of symptoms. Pirin (PIR) is involved in a variety of biological and molecular processes. However, the role of PIR in MASLD remains unclear. Rats were fed with high-fat and high-cholesterol (HFC) diet to induce MASLD. Time-course results showed that liver in HFC-fed rats exhibited a progressive increase in hepatic tissue damage over time (3, 6, and 9 weeks), and developed in obvious hepatic fibrosis and steatosis at week 9. Meanwhile, HFC feeding also gradually increased PIR expression in liver. In vitro , PIR expression was up-regulated in palmitic acid (PA)-induced Hep3B cells. PIR knockdown using PIR shRNA plasmid inhibited steatosis and expression of ACLY, FASN, and XBP1 in PA-induced Hep3B cells. Knocking down PIR inhibited hepatocyte inflammation by inhibiting phosphor-NF-κB p65 into the nucleus, which inhibited the expression of TNF-α and IL-1β. In MASLD, hepatocyte inflammation activates hepatic stellate cell, thereby leading to hepatic fibrosis. Supernatant from PA-treated Hep3B with PIR knockdown inhibited LX-2 activation, as evidenced by decreased expression of Col1A1 and α-SMA in LX-2 cells. Together, these results suggested that PIR knockdown might alleviate the progression of MASLD by inhibiting liver lipid deposition and inflammation, further inhibiting liver fibrosis. Transcriptome data analysis suggested that alteration in the expression of lipid metabolism-related pathways and genes might be a potential cause of PIR regulation in MASLD progression. These findings may reveal a novel target for MASLD therapy.
研究领域:
分子生物学
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肝病学
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代谢疾病
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细胞生物学
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生物信息学
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产品关联
DOI
PubMed
高影响因子
TOXICOLOGY AND APPLIED PHARMACOLOGY
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2023
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IF 3.8
Hancheng Huang,Shuchun Wei,Xiaohan Wu,Mengke Zhang,Benhong Zhou,Debin Huang,Weiguo Dong
The pathophysiological mechanism of hepatic fibrosis (HF) is related to the excessive activation of the DNA repair enzyme poly ADP-ribose polymerase-1 (PARP-1). The drugs, targeting PARP-1, are scarce. Therefore, the lead compound, moderately inhibiting PARP-1, with anti-HF properties should be identified. This study screened dihydrokaempferol (DHK) from herbs based on preliminary studies to intervene in a CCl 4 -induced liver injury and HF model in mice. In vitro , the expression levels of PARP-1-regulated related proteins and phosphorylation were examined. The binding pattern of DHK and PARP-1 was analyzed using molecular docking and molecular dynamics platforms. The results showed that DHK could significantly attenuate CCl 4 -induced liver injury and HF in mice. Moreover, it could also attenuate the toxic effects of CCl 4 on HepG2 and inhibit α-SMA and Collagen 1/3 synthesis of LX-2 cells in-vitro . Molecular docking revealed that DHK could competitively bind to the Glu-988 and His-862 residues of the upstream DNA repair enzyme PARP-1, moderately inhibiting its overactivation. This led to maintaining NAD + levels and energy metabolism in hepatocytes and inhibiting the activation of PARP-1-regulated downstream signaling pathways (TGF-β1, etc. ), related proteins (p-Smd2/3, etc. ), and inflammatory mediators while acting indirectly. Thus, DHK could attenuate CCl 4 -induced liver injury and HF in mice in a different mechanism from those of the existing reported flavonoids. It was associated with inhibiting the expression of downstream pathways and related cytokines by competitively binding to PARP-1. This study might provide a basis and direction for the design and exploration of anti-HF lead compounds.
研究领域:
药理学
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肝病学
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分子生物学
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毒理学
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产品关联
DOI
PubMed
高影响因子
MedComm
·
2023
·Vol. 4
·期 5
·e354
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IF 9.9
Yina Zhang,Liying Wang,Jiajia Shao,Yanning Liu,Yining Lu,Jing Yang,Siduo Xu,Jingkang Zhang,Minwei Li,Xiangrui Liu,Min Zheng
Calcipotriol (CAL) has been widely studied as a fibrosis inhibitor and used to treat plaque psoriasis via transdermal administration. The clinical application of CAL to treat liver fibrosis is bottlenecked by its unsatisfactory pharmacokinetics, biodistribution, and side effects, such as hypercalcemia in patients. The exploration of CAL as a safe and effective antifibrotic agent remains a major challenge. Therefore, we rationally designed and synthesized a self-assembled drug nanoparticle encapsulating CAL in its internal hydrophobic core for systematic injection (termed NPs/CAL) and further investigated the beneficial effect of the nanomaterial on liver fibrosis. C57BL/6 mice were used as the animal model, and human hepatic stellate cell line LX-2 was used as the cellular model of hepatic fibrogenesis. Immunofluorescence staining, flow cytometry, western blotting, immunohistochemical staining, and in vitro imaging were used for evaluating the efficacy of NPs/CAL treatment. We found NPs/CAL can be quickly internalized in vitro, thus potently deactivating LX-2 cells. In addition, NPs/CAL improved blood circulation and the accumulation of CAL in liver tissue. Importantly, NPs/CAL strongly contributed to the remission of liver fibrosis without inducing hypercalcemia. Overall, our work identifies a promising paradigm for the development of nanomaterial-based agents for liver fibrosis therapy.
研究领域:
药理学
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纳米技术
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肝病学
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药物递送系统
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产品关联
DOI
PubMed
高影响因子
SCANDINAVIAN JOURNAL OF GASTROENTEROLOGY
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2023
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IF 1.9
Wenchao Pan,Yadong Wang,Caiyan Zhao
Objective To explore the protective effect and related mechanism of miR-140-5p on liver fibrosis by interfering with TGF-β/Smad signaling pathway.Methods Liver fibrosis mice models were established by intraperitoneal injection of CCL4. Hematoxylin and eosin (HE) staining was used to detect the structural and morphological changes of the liver. Masson staining was used to detect collagen deposition. Human hepatic stellate cells (HSCs, LX-2) were transfected with miR-140-5p mimic or inhibitor then treated with TGF-β1. The qRT-PCR and Western blotting was used to detect the expression of related molecules. The luciferase reporter assay was used to identify the target of miR-140-5p.Results Our results indicated that miR-140-5p expression was downregulated in fibrotic liver tissues of model mice and LX-2 cells treated with TGF-β1. The overexpression of miR-140-5p decreased the expression of collagen1(COL1) and α-smooth muscle actin(α-SMA), inhibited the phosphorylation of Smad-2/3 (pSmad-2/3) in LX-2 cells. Conversely, the knockdown of miR-140-5p upregulated COL1 and α-SMA expression, increased Smad-2/3 phosphorylation. A dual-luciferase reporter assay showed that TGFβR1 was a target gene of miR-140-5p. The overexpression of miR-140-5p suppressed TGFβR1 expression in LX-2 cells. Additionally, knockdown of TGFβR1 decreased the expression of COL1 and α-SMA. Conversely, the overexpression of TGFβR1 reversed the inhibitory effect of miR-140-5p upregulation on expression of COL1 and α-SMA.Conclusion miR-140-5p bound to TGFβR1 mRNA 3′-untranslated region(3′UTR) and inhibited the expression of TGFβR1, pSmad-2/3, COL1 and α-SMA, thereby exerting a potential therapeutic effect on hepatic fibrosis.
研究领域:
分子生物学
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肝病学
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药理学
✓ 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）