EXPERIMENTAL PHYSIOLOGY
·
2020
·Vol. 105
·期 11
·1918-1927
·
IF 2.431
Manfeng Wang,Xiaowei Wu,Lu Gan,Zongyan Teng,Haijin Zhang,Yina Zhang
New Findings What is the central question of this study? Does Dnmt3a play a crucial role in regulating diabetic muscle atrophy? What is the main finding and its importance? Muscle atrophy is one of the major long-term complications of diabetes mellitus. However, little is known about the molecular mechanism involved. In this paper, we demonstrated that Dnmt3a overexpression effectively improves the diabetic muscle health in mice and documented the underlying mechanisms. DNMT3A might become a promising target to prevent muscle atrophy in patients with diabetes. Muscle atrophy is one of the major long-term complications of diabetes mellitus, which greatly affects the mobility of patients. Epigenetic processes mediated by DNA methyltransferases (DNMTs) play crucial roles in the locomotor system, but little is known about the functions of DNMTs in diabetic muscle atrophy. Here, we investigated the function of Dnmt3a in diabetic muscle atrophy and explored the mechanisms involved. Adeno-associated virus AAV2 overexpressing Dnmt3a or its vector control was injected into the tibialis anterior muscle of streptozotocin-induced diabetic mice. Muscle mass and muscle cross-sectional area were used to evaluate muscle atrophy. In vitro , adeno-associated virus AAV2 overexpressing Dnmt3a or its vector control was transfected into C2C12 myoblasts. Horse serum was used to induce differentiation and palmitate to stimulate the C2C12 myoblasts. The expressions of myogenic regulatory factors were examined by real-time PCR and western blot analysis. Overexpression of Dnmt3a attenuated muscle atrophy in diabetic mice and promoted myotube formation of C2C12 myoblasts. Overexpression of Dnmt3a restored the expressions of myogenic regulatory factors atrogin-1, MuRF1, Pax7, Myod1 and myogenin, both in vivo and in vitro . Moreover, overexpression of Dnmt3a activated the phosphorylation of Akt by inhibiting the activation of Pten. This study demonstrates that overexpression of Dnmt3a prevents diabetic muscle atrophy by modulating the Pten/Akt pathway.
研究领域:
分子生物学
·
糖尿病
·
肌肉生理学
·
表观遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2021
·
IF 6.291
Yuan Liu,Yaxin Yao,Wenjing Tao,Feng Liu,Songbai Yang,Ayong Zhao,Dan Song,Xiangchen Li
Bisphenol A (BPA) is a widely distributed environmental endocrine disruptor. The accumulation of BPA has been proved that produce various toxic effects both on human and animals. However, the strategies to reduce the damage of BPA on the body and related mechanisms remain to be studied. Coenzyme Q10 (CoQ10), as a powerful antioxidant, is ubiquitous in many eukaryotic cells, which can improve the integrity of lysosomal membrane, lysosomal degradation function and promote autophagy. Here, we examined the ability of CoQ10 to alleviate oxidative stress and apoptosis in BPA-induced damages in C2C12 cells, and how to alleviate it. Our results showed that BPA treatment significantly reduced cell viability, increased the number of cell apoptosis and ROS production, decreased mitochondrial membrane potential, and inhibited the gene expression of mitochondria biogenesis. Moreover, we demonstrated that exposure to BPA increased expression levels of autophagy protein (LC3-II, p62), inhibited autophagy flux, and disrupted the acidic pH environment of lysosomes. Importantly, CoQ10 supplementation effectively restored these abnormalities caused by BPA. CoQ10 significantly decreased the apoptotic incidence and ROS levels, improved mitochondrial membrane potential. Moreover, CoQ10 improved lysosome function and enhanced autophagy flux. Taken together, our results indicate that CoQ10 supplementation is a feasible and effective way to promote the level of autophagy by improving lysosomal function, thereby reducing the apoptosis caused by BPA accumulation. This study aims to provide evidence for the role of CoQ10 in repairing BPA-induced cell damage in clinical practice.
研究领域:
毒理学
·
细胞生物学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Materials
·
2019
·Vol. 12
·期 20
·3391
·
IF 2.972
Zheng Wang,Xiaolu Zhu,Ruiyuan Zhang
The interactions between substrate materials and cells usually play an important role in the hydrogel-based 3D cell cultures. However, the hydrogels that are usually used could not be parametrically regulated, especially for quantitatively regulating the spatial distribution of the adhesion sites for cells in 3D. Here, we employed the semisynthetic hydrogel consisting of maleimide-dextran, Arg-Gly-Asp (RGD) peptides, and cell degradable crosslinkers to biochemically characterize the evolutionary behaviors of NIH–3T3 fibroblasts and C2C12 cells in 3D. Moreover, by comparing the cell-adhesive efficacy of 3D dextran hydrogels with four different RGD clustering rates, we explored the underlying regulation law of C2C12 connections and 3T3 aggregations. The results showed that mal-dextran hydrogel could promise cells stable viability and continuous proliferation, and induce more self-organized multicellular structures relative to 2D culture. More importantly, we found that RGD-clustered mal-dextran hydrogel has the advantage of enhancing C2C12 cell elongation and the breadthwise-aggregated connection, and promoting the 3T3 cell aggregating degree compared to that with homogenous RGD. Further, the advantages of RGD clustering hydrogel could be amplified by appropriately reducing RGD concentration. Such RGD-composition controllable mal-dextran hydrogel can function as a regulator of the collective cellular behaviors, which provides useful information for quantitatively designing the tailored hydrogel system and exploiting advanced biomaterials.Keywords:glycan;maleimide-dextran hydrogel;cell–substrate adhesion;3D culture
研究领域:
生物材料
·
细胞生物学
·
组织工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Drug Design Development and Therapy
·
2022
·
IF 4.319
Xiangtian Ling,Xinqi Ma,Xielan Kuang,Yuxiu Zou,Han Zhang,Han Tang,Han Du,Binbin Zhu,Hao Huang,Qing Xia,Minghao Chen,Danyi Mao,Dongli Chen,Huangxuan Shen,Jianhua Yan
Purpose To assess the cellular and molecular effects of lidocaine on muscles/myoblasts.Methods Cultured myogenic precursor (C2C12) cells were treated with varying concentrations of lidocaine.Results Cell viability of C2C12 cells was inhibited by lidocaine in a concentration-dependent manner, with concentrations ≥0.08%, producing a dramatic reduction in cell viability. These ≥0.08% concentrations of lidocaine arrested cell cycles of C2C12 cells in the G0/G1 phase. Moreover, lidocaine inhibited cell migration and myogenic processes in C2C12 cells at low concentrations. Results from QRT-PCR assays revealed that following treatment with lidocaine, Notch1, Notch2, Hes1, Csl and Dll4 all showed higher levels of expression, while no changes were observed in Mmal1, Hey1, Dll1 and Jag1.Conclusion This work provides the first description of the effects of lidocaine upon the regeneration of muscles and maintenance of satellite cells at the cellular and molecular levels. In specific, we found that the Dll4-Notch-Csl-Hes1 axis was up-regulated suggesting that the Notch signaling pathway was involved in producing these effects of lidocaine. These findings provide a new and important foundation for future investigations into the effects of drug therapies in muscle diseases.
研究领域:
药理学
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2017
·
IF 7.504
Serge Ostrovidov,Majid Ebrahimi,Hojae Bae,Hung Kim Nguyen,Sahar Salehi,Sang Bok Kim,Akichika Kumatani,Tomokazu Matsue,Xuetao Shi,Ken Nakajima,Shizu Hidema,Makoto Osanai,Ali Khademhosseini
In this study, composite gelatin–polyaniline (PANI) nanofibers doped with camphorsulfonic acid (CSA) were fabricated by electrospinning and used as substrates to culture C2C12 myoblast cells. We observed enhanced myotube formation on composite gelatin–PANI nanofibers compared to gelatin nanofibers, concomitantly with enhanced myotube maturation. Thus, in myotubes, intracellular organization, colocalization of the dihydropyridine receptor (DHPR) and ryanodine receptor (RyR), expression of genes correlated to the excitation–contraction (E–C) coupling apparatus, calcium transients, and myotube contractibility were increased. Such composite material scaffolds combining topographical and electrically conductive cues may be useful to direct skeletal muscle cell organization and to improve cellular maturation, functionality, and tissue formation.
研究领域:
生物材料
·
组织工程
·
肌肉生理学
·
生物医学工程
✓ 已验证
DOI
PubMed
高影响因子
BIOMATERIALS
·
2014
·
IF 8.312
Serge Ostrovidov,Xuetao Shi,Ling Zhang,Xiaobin Liang,Sang Bok Kim,Toshinori Fujie,Murugan Ramalingam,Mingwei Chen,Ken Nakajima,Faten Al-Hazmi,Hojae Bae,Adnan Memic,Ali Khademhosseini
Engineering functional muscle tissue requires the formation of densely packed, aligned, and mature myotubes. To enhance the formation of aligned myotubes with improved contractibility, we fabricated aligned electrospun gelatin multi-walled carbon nanotubes (MWNTs) hybrid fibers that were used as scaffolds for the growth of myoblasts (C2C12). The MWNTs significantly enhanced myotube formation by improving the mechanical properties of the resulting fibers and upregulated the activation of mechanotransduction related genes. In addition, the fibers enhanced the maturation of the myotubes and the amplitude of the myotube contractions under electrical stimulation (ES). Such hybrid material scaffolds may be useful to direct skeletal muscle cellular organization, improve cellular functionality and tissue formation.
研究领域:
组织工程
·
生物材料
·
细胞生物学
·
肌肉骨骼研究
✓ 已验证
DOI
PubMed
高影响因子
Molecular Therapy-Nucleic Acids
·
2023
·
IF 8.8
Xinyi Gu,Shen Wang,Dongdong Li,Bo Jin,Zhidan Qi,Jin Deng,Chen Huang,Xiaofeng Yin
Peripheral nerve injury can lead to progressive muscle atrophy and poor motor function recovery, which is a difficult point of treatment, and the mechanism needs to be further explored. In previous studies we found that miR-142a-3p was significantly upregulated and persistently high expressed in denervated mouse skeletal muscle. Here, we show that overexpression of miR-142a-3p inhibited the growth and differentiation of C2C12 myoblast, while knockdown of miR-142a-3p had a promoting effect. In vitro, knockdown of miR-142a-3p in denervated mouse skeletal muscle effectively increased proliferating muscle satellite cells and ameliorated muscle atrophy. Mechanistically, the myoregulator Mef2a was proved to be an important downstream target of miR-142a-3p, and miR-142a-3p regulates skeletal muscle differentiation and regeneration by inhibiting the expression of Mef2a. The co-knockdown of Mef2a and miR-142a-3p effectively alleviated or offset the biological effects of miR-142a-3p knockdown. In conclusion, our data revealed that miR-142a-3p regulates neurogenic skeletal muscle atrophy by targeting Mef2a.
研究领域:
神经科学
·
分子生物学
·
肌肉骨骼疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
STAR Protocols
·
2024
·
IF 1.3
Zili Gao,Shenglong Ding,Tingting Fan,Wenhui Huang,Xiyuan Zhao,Xin Liu,Wenli Liu,Mingzhu Zhang,Qi Gu
Summary Here, we present a protocol for 3D printing heart tissues using thiol-norbornene photoclick collagen (NorCol). We describe steps for synthesizing NorCol, preparing bioink and the support bath, and cell-laden printing. We then detail procedures for the loading of C2C12 cells into NorCol, ensuring structural integrity and cell viability after printing. This protocol is adaptable to various cell lines and allows for the printing of diverse complex structures, which can be used in drug screening and disease modeling.
研究领域:
生物医学工程
·
组织工程
·
材料科学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Biomaterials Science & Engineering
·
2020
·
IF 4.152
Honglei Wang,Xue Qu,Zheng Zhang,Miao Lei,Haoqi Tan,Chunyan Bao,Shaoliang Lin,Linyong Zhu,Joachim Kohn,Changsheng Liu
The construction of a biomaterial matrix with biological properties is of great importance to developing functional materials for clinical use. However, the site-specific immobilization of growth factors to endow materials with bioactivities has been a challenge to date. Considering the wide existence of glycosylation in mammalian proteins or recombinant proteins, we establish a bioaffinity-based protein immobilization strategy (bioanchoring method) utilizing the native sugar–lectin interaction between concanavalin A (Con A) and the oligosaccharide chain on glycosylated bone morphogenetic protein-2 (GBMP-2). The interaction realizes the site-specific immobilization of GBMP-2 to a substrate modified with Con A while preserving its bioactivity in a sustained and highly efficient way, as evidenced by its enhanced ability to induce osteodifferentiation compared with that of the soluble GBMP-2. Moreover, the surface with Con A-bioanchored GBMP-2 can be reused to stimulate multiple batches of C2C12 cells to differentiate almost to the same degree. Even after 4 month storage at 4 °C in phosphate-buffered saline (PBS), the Con A-bioanchored GBMP-2 still maintains the bioactivity to stimulate the differentiation of C2C12 cells. Furthermore, the ectopic ossification test proves the in vivo bioactivity of bioanchored GBMP-2. Overall, our results demonstrate that the tag-free and site (i.e., sugar chain)-specific protein immobilization strategy represents a simple and generic alternative, which is promising to apply for other glycoprotein immobilization and application. It should be noted that although the lectin we utilized can only bind to d-mannose/d-glucose, the diversity of the lectin family assures that a specific lectin could be offered for other sugar types, thus expanding the applicable scope further.
研究领域:
生物材料
·
组织工程
·
生物医学工程
·
蛋白质工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Regenerative Biomaterials
·
2021
·Vol. 8
·期 5
·
IF 6.353
Wang Ying,Wang Qingshuai,Luo Shengchang,Chen Zhoujiang,Zheng Xiang,Kankala Ranjith Kumar,Chen Aizheng,Wang Shibin
Recently, hydrogels have gained enormous interest in three-dimensional (3D) bioprinting toward developing functional substitutes for tissue remolding. However, it is highly challenging to transmit electrical signals to cells due to the limited electrical conductivity of the bioprinted hydrogels. Herein, we demonstrate the 3D bioprinting-assisted fabrication of a conductive hydrogel scaffold based on poly-3,4-ethylene dioxythiophene (PEDOT) nanoparticles (NPs) deposited in gelatin methacryloyl (GelMA) for enhanced myogenic differentiation of mouse myoblasts (C2C12 cells). Initially, PEDOT NPs are dispersed in the hydrogel uniformly to enhance the conductive property of the hydrogel scaffold. Notably, the incorporated PEDOT NPs showed minimal influence on the printing ability of GelMA. Then, C2C12 cells are successfully encapsulated within GelMA/PEDOT conductive hydrogels using 3D extrusion bioprinting. Furthermore, the proliferation, migration and differentiation efficacies of C2C12 cells in the highly conductive GelMA/PEDOT composite scaffolds are demonstrated using various in vitro investigations of live/dead staining, F-actin staining, desmin and myogenin immunofluorescence staining. Finally, the effects of electrical signals on the stimulation of the scaffolds are investigated toward the myogenic differentiation of C2C12 cells and the formation of myotubes in vitro. Collectively, our findings demonstrate that the fabrication of the conductive hydrogels provides a feasible approach for the encapsulation of cells and the regeneration of the muscle tissue.
研究领域:
组织工程
·
生物材料
·
细胞生物学
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Jing Xie,Li Zhang,Yu-Ying Bai,Wen-Jing Wang,Xia Hu,Shijun Li,Yang Tian
Walnut septum is rich in polyphenolic substances, but the hypoglycaemic and hypolipidemic effects of walnut septum polyphenols (WSP) and the main active compounds responsible for these effects are still unknown. In the present study, WSP was firstly extracted, and then purified using D101 macroporous resin, and the purity of WSP obtained was 87.36 %. WSP inhibited the activity of α-glucosidase and pancreatic lipase with IC 50 values of 36.2 μg/mL and 155.9 μg/mL, respectively. Furthermore, WSP increased glucose consumption and decreased oxidative stress in insulin-resistant C2C12 cells, as well as inhibited lipid accumulation in 3 T3-L1 adipocytes. Eight naturally active ligands for α-glucosidase and pancreatic lipase were identified from WSP by affinity ultrafiltration combined with liquid chromatography-mass spectrometry analytical methods. Further molecular docking showed strong binding between these eight ligands and the enzyme. The results of the study lay a scientific foundation for the development of walnut septum as a functional food for hypoglycemic and hypolipidemic.
研究领域:
食品科学
·
药理学
·
生物化学
·
营养学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
·
2023
·
IF 5.2
Lanting Wei,Xiang Wang,Junjie Fu,Jian Yin,Jing Hu
The current therapy for muscle injury is facing challenges in tissue self-regeneration and function restoration, which declines steeply with age. Tissue engineering is a promising strategy for treating skeletal muscular disorders. Natural polysaccharides have great potential to support cell growth and attachment as tissue-engineered scaffolds. A three-dimensional (3D) porous hydrogel scaffold has been prepared based on the double network of natural polysaccharide sodium alginate (SA), chitosan (CS) and Ca 2+ ions through the freeze-drying process. The physical, mechanical, and biological properties of the CS/SA/Ca 2+ hydrogel scaffold can be controlled by adjusting the SA/CS ratio. The physically cross-linked double network hydrogel showed good biosafety, high stability and satisfactory degradation rate . When the CS/SA molar ratio was 1:1.2 (H-1.2 group), the pore size and compression modulus of the double network hydrogel were 104.06 ± 2.64 µm and 40.00 ± 2.52 kPa, respectively. The results of in vitro evaluation showed that the 3D structure of the CS/SA/Ca 2+ double network hydrogel could well support adhesion and proliferation of C2C12 murine myoblasts and porcine skeletal muscle stem cells. The best cell growth effect was achieved in the H-1.2 group. The physically cross-linked CS/SA/Ca 2+ double network hydrogel can be a promising printable material as the cell scaffold in applications of skeletal muscle tissue engineering.
研究领域:
组织工程
·
生物材料
·
生物医学工程
·
细胞生物学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）