Food & Function
·
2020
·Vol. 12
·期 1
·144-153
·
IF 4.171
Shuge Liu,Yunmei Sun,Rui Zhao,Yingqian Wang,Wanrong Zhang,Weijun Pang
Isoleucine (Ile), as a branched-chain amino acid (BCAA), has a vital role in regulating body weight and muscle protein synthesis. However, the regulatory effect of Ile on muscle mass under high-fat diet (HFD) conditions and intramyocellular lipid deposition remains largely unclear. In this study, a feeding experiment with HFD with or without 25 g L−1 Ile was performed using 32 wild male C57BL/6J mice randomly divided into two groups. The results showed that Ile significantly increased both muscle and fat mass, as well as causing insulin resistance and meanwhile upregulating the levels of key adipogenic and myogenic proteins. More importantly, Ile damaged the mitochondrial function by vacuolation, swelling and cristae fracture in the gastrocnemius (GAS) and tibialis anterior (TA) with downregulation of mitochondrial function-related genes. Furthermore, Ile promoted myogenesis and more lipid droplet accumulation in myotubes. Compared with the control, the protein levels of myosin heavy chain (MyHC), myoblast determination protein 1 (MyoD), myogenin (MyoG), peroxisome proliferator-activated receptor gamma (PPARg) and fatty acid synthase (FAS) were upregulated in the Ile group, whereas the protein levels of adipose triglyceride lipase (ATGL) and lipoprotein lipase (LPL) were downregulated. Collectively, Ile increased muscle mass through myogenesis and intramyocellular lipid deposition. Our findings provide a new perspective for not only improving the lean juiciness of farm animals by increasing intramyocellular lipid accumulation, but also modulating myopathies under obesity.
研究领域:
营养学
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代谢学
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分子生物学
·
生理学
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产品关联
DOI
PubMed
高影响因子
FASEB JOURNAL
·
2023
·Vol. 37
·期 4
·e22866
·
IF 4.8
Shan-Yan Gao,Yong-Ping Liu,Ri Wen,Xin-Mei Huang,Ping Li,Yu-Hang Yang,Ni Yang,Tie-Ning Zhang
Uncontrolled diabetes causes a catabolic state with multi-organic complications, of which impairment on skeletal muscle contributes to the damaged mobility. Kcnma1 gene encodes the pore-forming α-subunit of Ca 2+ - and voltage-gated K + channels of large conductance (BK channels), and loss-of-function mutations in Kcnma1 are in regards to impaired myogenesis. Herein, we observed a time-course reduction of Kcnma1 expression in the tibialis anterior muscles of leptin receptor-deficient (db/db) diabetic mice. To investigate the role of Kcnma1 in diabetic muscle atrophy, muscle-specific knockdown of Kcnma1 was achieved by mice receiving intravenous injection of adeno-associated virus-9 (AAV9)-encoding shRNA against Kcnma1 under the muscle creatine kinase (MCK) promoter. Impairment on muscle mass and myogenesis were observed in m/m mice with AAV9-sh Kcnma1 intervention, while this impairment was more obvious in diabetic db/db mice. Simultaneously, damaged mitochondrial dynamics and biogenesis showed much severer in db/db mice with AAV9-sh Kcnma1 intervention. RNA sequencing revealed the large transcriptomic changes resulted by Kcnma1 knockdown, and changes in mitochondrial homeostasis-related genes were validated. Besides, the artificial alteration of Kcnma1 in mouse C2C12 myoblasts was achieved with an adenovirus vector. Consistent results were demonstrated by Kcnma1 knockdown in palmitate-treated cells, whereas opposite results were exhibited by Kcnma1 overexpression. Collectively, we document Kcnma1 as a potential keeper of mitochondrial homeostasis, and the loss of Kcnma1 is a critical event in priming skeletal muscle loss in diabetes.
研究领域:
糖尿病
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肌肉生物学
·
分子遗传学
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细胞生理学
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DOI
PubMed
高影响因子
Cell Death & Disease
·
2021
·Vol. 12
·期 12
·1-13
·
IF 8.469
Tan Yanjie,Jin Yi,Zhao Pengxiang,Wu Jian,Ren Zhuqing
Lipid droplet (LD), a multi-functional organelle, is found in most eukaryotic cells. LDs participate in the regulation of many cellular processes including proliferation, stress, and apoptosis. Previous studies showed the athlete’s paradox that trained athletes accumulate LDs in their skeletal muscle. However, the impact of LDs on skeletal muscle and myogenesis is not clear. We discovered that C2C12 myoblast cells containing more LDs formed more multinucleated muscle fibers. We also discovered that LDs promoted cell migration and fusion by promoting actin-filaments remodeling. Mechanistically, two LD-proteins, Acyl-CoA synthetase long chain family member 3 (ACSL3) and lysophosphatidylcholine acyltransferase 1 (LPCAT1), medicated the recruitment of actinin proteins which contributed to actin-filaments formation on the surface of LDs. During remodeling, the actinin proteins on LDs surface translocated to actin-filaments via ARF1/COPI vesicles. Our study demonstrate LDs contribute to cell differentiation, which lead to new insight into the LD function.
研究领域:
细胞生物学
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分子生物学
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肌肉生理学
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产品关联
DOI
PubMed
高影响因子
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
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肌肉生理学
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表观遗传学
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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
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
FISH & SHELLFISH IMMUNOLOGY
·
2023
·
IF 4.7
Ju Li,Jinmei Zuo,Xiaoyan Lv,Jiahe Ma,Xiaohong Li,Simiao Fu,Jinsheng Sun
Tissue autotomy is a unique adaptive response to environmental stress, followed by regeneration process compensating for the loss of body parts. The crustaceans present remarkable activity of appendage autotomy and regeneration, however, the molecular mechanism is still unclear. In this study, the Eriocheir sinensis Hedgehog ( EsHH ) and Smoothened ( EsSMO ) were identified in the regenerative limbs, and the function of Hedgehog signaling pathway on limb regeneration was evaluated. At the blastema growth stage of limb regeneration, the expression of EsHH and EsSMO was up-regulated in response to limb autotomy stress, and down-regulated at blastema differentiation stage. To clarify the effect of Hedgehog pathway during limb regeneration, the regenerative efficiency was evaluated with Smoothened inhibitor cyclopamine or RNAi ( ds-HH ) injection. We observed that the regenerative efficiency was significantly repressed with blockage of Hedgehog pathway at both the basal growth stage and the proecdysial growth stage, which was indicated by the delay of wound healing and blastema growth, as well as a decrease in the size of newly formed limbs. In addition, gene expression and BrdU incorporation assay showed that the proliferation and myogenic differentiation of blastema cells were suppressed with either cyclopamine or ds- HH injection. Thus, these results suggest that Hedgehog signaling pathway is essential for the establishment of limb regeneration in E. sinensis through promoting the proliferation and myogenic differentiation of blastema cells.
研究领域:
发育生物学
·
分子生物学
·
再生医学
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产品关联
DOI
PubMed
高影响因子
eLife
·
2023
·
IF 7.7
Zhao-Bo Luo,Shengzhong Han,Xi-Jun Yin,Hongye Liu,Junxia Wang,Meifu Xuan,Chunyun Hao,Danqi Wang,Yize Liu,Shuangyan Chang,Dongxu Li,Kai Gao,Huiling Li,Biaohu Quan,Lin-Hu Quan,Jin-Dan Kang
The host genome may influence the composition of the intestinal microbiota, and the intestinal microbiota has a significant effect on muscle growth and development. In this study, we found that the deletion of the myostatin (MSTN) gene positively regulates the expression of the intestinal tight junction-related genes TJP1 and OCLN through the myosin light-chain kinase/myosin light chain pathway. The intestinal structure of MSTN−/− pigs differed from wild-type, including by the presence of a thicker muscularis and longer plicae. Together, these changes affect the structure of intestinal microbiota. Mice transplanted with the intestinal microbiota of MSTN−/− pigs had myofibers with larger cross-sectional areas and higher fast-twitch glycolytic muscle mass. Microbes responsible for the production of short-chain fatty acids (SCFAs) were enriched in both the MSTN−/− pigs and recipient mice, and SCFAs levels were elevated in the colon contents. We also demonstrated that valeric acid stimulates type IIb myofiber growth by activating the Akt/mTOR pathway via G protein-coupled receptor 43 and ameliorates dexamethasone-induced muscle atrophy. This is the first study to identify the MSTN gene-gut microbiota-SCFA axis and its regulatory role in fast-twitch glycolytic muscle growth.
研究领域:
遗传学
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微生物学
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肌肉生物学
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胃肠病学
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产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2024
·
IF 4.8
Yawen Chen,Wenjing Dai,Wanli Xiong,Yi Zhou,Ziying Chen,Chengxin Ge,Yan Zhou
As a meat substitute, cell cultured meat aims to simulate the composition of real meat to provide nutrient substance. The myotube formation efficiency of myoblasts in vitro is the key for simulating the structure and composition of muscle tissue. Therefore, this study is aimed to explore the most effective groove widths for myoblasts fusion and related molecular mechanism. The results of immunofluorescence showed that the grooves with a width range of 50 μm–1000 μm were beneficial to cell arrangement compared to flat structure. At the same time, the expression of myogenin ( Myog ) and myosin heavy chain 2 ( Myh2 ) up-regulated. However, the cell fusion index of myoblasts was higher in grooves with a width of 100 μm, 200 μm and 300 μm. Furthermore, Cytochalasin D disrupted intracellular stress fibers and myotube formation. However, there were still many myotubes formed in the 200 μm groove with the addition of Cytochalasin D. The expression of fusion related genes was higher than the flat group. Moreover, incorporating groove structure into scaffolds of soybean protein isolate and pea protein isolate could enhance cell adhesion, activity and myogenic differentiation of myoblasts. This could provide ideas for scaffolds design of cultured meat products using groove structure.
研究领域:
组织工程
·
细胞生物学
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生物技术
·
食品科学
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DOI
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
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产品关联
DOI
PubMed
高影响因子
LIFE SCIENCES
·
2023
·
IF 6.1
Pan He,Guanhua Du,Xuemei Qin,Zhenyu Li
Aims Sarcopenia is an age-related syndrome characterized by a gradual loss of the muscle mass, strength, and function. It is associated with a high risk of adverse consequences such as poorer quality of life, falls, disability and mortality among the elderly. The aim in this study is to investigate the pathological mechanism of sarcopenia. Main methods The aging of skeletal muscle was investigated by the D-galactose induced accelerated aging model combining with constrained motion. After 10 weeks, muscle function and gastrocnemius muscle index, and morphology of muscle fibers were evaluated, and myostatin, IGF-1 and ATP in skeletal muscle were also determined. Then the mechanism of aging-related skeletal muscle dysfunctions was investigated based on untargeted serum metabolomics and 16S rRNA gene sequencing. Four key metabolites were validated by the D-galactose-induced C2C12 senescent cell model in vitro . Key findings Results showed that gastrocnemius muscle mass was decreased significantly, morphology of muscle fibers was altered, and muscle function was damaged in the aged group. Furthermore, increased MSTN, and decreased IGF-1 and ATP were also observed in the aging skeletal muscle. Importantly, alteration of the key pathways including riboflavin biosynthesis and energy metabolism contributed to the aging of skeletal muscle. Four key metabolites, including riboflavin, α-ketoglutaric acid and two dicarboxylic acids, which were involved in these metabolic pathways, could promote the proliferation of C2C12 cells. Significance These findings provide novel insights into pathological mechanism of sarcopenia, and will facilitate the development of therapeutic and preventive strategies for sarcopenia.
研究领域:
老年学
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肌肉生理学
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代谢组学
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微生物组研究
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产品关联
DOI
PubMed
高影响因子
Chem
·
2025
·
IF 19.6
Zhiwen Luo,Renwen Wan,Jiajun Qiu,Chen Chen,Yisheng Chen,Zhijie Zhao,Jie Mei,Yanwei He,Xinting Feng,Zhen Peng,Siwei Yin,Mowen Xu,Zhiheng Lin,James Hoipo Hui,Xuanyong Liu,Junming Sun,Shiyi Chen,Yaying Sun
Skeletal muscle inherently possesses regenerative capabilities, but recovery from severe volumetric muscle loss (VML) is often constrained, potentially resulting in loss of muscular functionality. Effective biomaterial interventions are rare. To address these gaps, we developed a novel injectable hydrogel, SiCP@InjGel, which utilizes in situ self-assembly to combine puerarin, chitosan, and mesoporous silica nanoparticles, targeting skeletal muscle repair. This hydrogel was rigorously tested in mice, rat, and porcine VML models through in vitro and in vivo experiments, demonstrating its ability to modulate the immune microenvironment, enhance myogenesis, and improve functional muscle recovery. Utilizing multiple state-of-the-art omics technologies, including mass cytometry and single-cell RNA sequencing, we identified that it increased Pdpn+ macrophages and MyoG+ muscle stem cells, reshaping the immune landscape and accelerating muscle regeneration at molecular and cellular resolutions. These findings establish SiCP@InjGel as a multifunctional biomaterial platform with significant translational potential for clinical management of severe skeletal muscle injuries.
研究领域:
再生医学
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生物材料科学
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免疫学
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分子生物学
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组织工程
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系统生物学
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转化医学
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DOI
高影响因子
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
·
2023
·
IF 2.1
Ke Shenghui,Feng Yiwei,Luo Liying,Qin Wanzhao,Liu Huayu,Nie Jingchong,Liang Beijiang,Ma Hongjie,Xie Mao,Li Jingyu,Niu Zhijie,Li Guojian,Tang Anzhou,Xia Wei,He Guangyao
A method for the in vitro isolation, purification, identification, and induced differentiation of satellite cells from adult tree shrew skeletal muscle was established. The mixed enzyme digestion method and differential adhesion method were used to obtain skeletal muscle satellite cells, which were identified and induced to differentiate to verify their pluripotency. The use of a mixture of collagenase II, hyaluronidase IV, and DNase I is an efficient method for isolating adult tree shrew skeletal muscle satellite cells. The P3 generation of cells had good morphology, rapid proliferation, high viability, and an “S”-shaped growth curve. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining indicated that marker genes or proteins were expressed in skeletal muscle satellite cells. After myogenic differentiation was induced, multiple-nucleated myotubes were observed, and the MyHC protein was expressed. The expression of myogenic marker genes changed with the differentiation process. After the induction of adipogenic differentiation, orange-red lipid droplets were observed, and the expression of adipogenic marker genes increased gradually with the differentiation process. In summary, satellite cells from adult tree shrew skeletal muscle were successfully isolated using a mixed enzyme digestion method, and their potential for differentiation into myogenic and adipogenic cells was confirmed, laying a foundation for further in vitro study of tree shrew muscle damage.
研究领域:
细胞生物学
·
肌肉生理学
·
干细胞研究
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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）