Complete skeletal muscle repair and regeneration due to severe large injury or disease is still a challenge.Biochemical cues are critical to control myoblast cell function and can be utilized to develop smart biomater...Complete skeletal muscle repair and regeneration due to severe large injury or disease is still a challenge.Biochemical cues are critical to control myoblast cell function and can be utilized to develop smart biomaterials for skeletal muscle engineering.Citric acid-based biodegradable polymers have received much attention on tissue engineering,however,their regulation on myoblast cell differentiation and mechanism was few investigated.Here,we find that citrate-based polycitrate-polyethylene glycol-polyethylenimine(POCG-PEI600)nanoclusters can significantly enhance the in vitro myoblast proliferation by probably reinforcing the mitochondrial number,promote the myotube formation and full-thickness skeletal muscle regeneration in vivo by activating the myogenic biomarker genes expression of Myod and Mhc.POCG-PEI600 nanoclusters could also promote the phosphorylation of p38 in MAP kinases(MAPK)signaling pathway,which led to the promotion of the myoblast differentiation.The in vivo skeletal muscle loss rat model also confirmed that POCG-PEI600 nanoclusters could significantly improve the angiogenesis,myofibers formation and complete skeletal muscle regeneration.POCG-PEI600 nanocluster could be also biodegraded into small molecules and eliminated in vivo,suggesting their high biocompatibility and biosafety.This study could provide a bioactive biomaterial-based strategy to repair and regenerate skeletal muscle tissue.展开更多
旨在了解m6A甲基转移酶样蛋白16(methyltransferase-like protein 16,METTL16)在鸡不同类型肌肉组织中的表达情况及其在鸡骨骼肌功能中的调控作用。本研究利用qRT-PCR技术检测METTL16基因在120日龄广西麻鸡母鸡不同类型肌肉组织中的表达...旨在了解m6A甲基转移酶样蛋白16(methyltransferase-like protein 16,METTL16)在鸡不同类型肌肉组织中的表达情况及其在鸡骨骼肌功能中的调控作用。本研究利用qRT-PCR技术检测METTL16基因在120日龄广西麻鸡母鸡不同类型肌肉组织中的表达,同时利用siRNA干扰鸡原代成肌细胞中METTL16基因的表达,分析其对成肌细胞增殖、分化和肌纤维形成的影响。结果显示,METTL16基因在鸡的不同类型肌肉组织(胸大肌、胸小肌、缝匠肌、耻坐骨肌内侧肌、耻坐骨肌外侧肌、髂胫外侧肌、腓肠肌内侧肌和背阔肌)中广泛表达,其中,在白肌纤维为主的胸大肌、胸小肌和髂胫外侧肌中表达量相对更高;细胞增殖检测结果显示,干扰METTL16基因表达后,鸡成肌细胞的增殖活力受到了抑制;在成肌细胞分化后干扰METTL16基因表达,鸡成肌细胞RNA m6A甲基化水平呈下降趋势,细胞分化关键基因MyoD表达显著下降(P<0.05),fast-MyHC基因表达显著上升(P<0.001),slow-MyHC基因表达呈下降趋势。综上,METTL16基因在不同类型肌肉中的表达与其肌纤维类型组成相关,METTL16基因可能在鸡成肌细胞增殖和分化中发挥重要作用,并抑制快肌纤维的形成,提示METTL16可能在鸡肌肉功能调控中发挥重要作用。展开更多
Sirt1(Sirtuin type 1)是依赖于烟酰胺腺嘌呤二核苷酸(NAD+)的组蛋白脱乙酰酶,为Sirtuins家族成员之一,与细胞增殖、分化、衰老、凋亡和代谢密切相关。目前,有关Sirt1与衰老和代谢的论文已在Science、Nature、Cell等杂志上连续刊出。其...Sirt1(Sirtuin type 1)是依赖于烟酰胺腺嘌呤二核苷酸(NAD+)的组蛋白脱乙酰酶,为Sirtuins家族成员之一,与细胞增殖、分化、衰老、凋亡和代谢密切相关。目前,有关Sirt1与衰老和代谢的论文已在Science、Nature、Cell等杂志上连续刊出。其中,Sirt1通过抑制PPARγ促进白色脂肪细胞中脂肪动员,并且通过下调肌细胞标志基因表达来抑制成肌细胞分化。提示Sirt1不仅是一个重要的与机体“长寿”有关的因子,而且可能在动物脂肪沉积和肌肉发育中起着关键的调控作用。展开更多
Growth and differentiation factor-11 (GDF-11) is a secreted protein that is closely related to myostatin, a known inhibitor of skeletal muscle development. The role of GDF-11 in regulating skeletal muscle growth remai...Growth and differentiation factor-11 (GDF-11) is a secreted protein that is closely related to myostatin, a known inhibitor of skeletal muscle development. The role of GDF-11 in regulating skeletal muscle growth remains unclear and the pattern of expression during post-natal growth has not been reported. Therefore, we sought to determine the expression of GDF-11 during post-natal growth and its effect on myoblast proliferation and differentiation. We collected gastrocnemius muscles from male and female mice at 2, 3, 4, 6, 12, 20 and 32 weeks of age (n = 6 per sex and age). In addition, gastrocnemius muscles were col- lected from male wild-type and myostatin knockout mice at 4, 6, 12 and 20 weeks of age (n = 6 per age and genotype). RNA was extracted and reverse tran- scribed. Northern analysis identified an expected 4.4 kb mRNA transcript for GDF-11 in gastrocnemius muscles of mice. The concentration of GDF-11 mRNA, as determined by quantitative PCR, was increased in gastrocnemius muscles from 2 to 6 weeks—a period of rapid postnatal muscle growth—and remained higher in male than female mice from 4 to 20 weeks of age (P gastrocnemius muscles of myostatin knockout compared with wild-type mice (P < 0.05), which may suggest a compensatory mecha- nism for the lack of myostatin. In support, recombi- nant GDF-11 inhibited differentiation of C2C12 mur- ine myoblasts and those isolated from myostatin knockout and wild-type mice (P < 0.05). Inhibited dif-ferentiation of C2C12 myoblasts was associated with decreased mRNA expression of early and late mo- lecular markers of differentiation (MyoD, myogenin, IGF-II, desmin and MyHC, P < 0.05). Our results suggest that GDF-11 regulates growth of skeletal muscles by inhibiting myoblast differentiation in an autocrine/paracrine manner and, perhaps, also plays a role in regulating sexually dimorphic growth.展开更多
Our previous work reported a relationship between FOXO1 mutations and growth of Qinchuan(QC) cattle. Here, we performed differential expression analysis of FOXO1 and its association analysis with growth traits in QC c...Our previous work reported a relationship between FOXO1 mutations and growth of Qinchuan(QC) cattle. Here, we performed differential expression analysis of FOXO1 and its association analysis with growth traits in QC cattle. First, we measured the expression of the FOXO1 gene in nine tissues during three developmental stages. The results showed that FOXO1 was abundantly expressed in tissues of calves but was strongly repressed in adulthood, although there was significant transcription in skeletal muscle. FOXO1 expression showed gradual up-regulation during differentiation of primary bovine skeletal muscle cells.We also identified six SNPs of the bovine FOXO1 gene by sequencing DNA pools of samples from 488 individuals, and association analysis indicated that five SNPs were significantly associated with some growth traits in the QC population. We further analyzed four haplotype combinations of the six SNPs and found significant correlation with body length(P<0.01). In conclusion, FOXO1 participates in bovine myocyte differentiation and expression, and may be a strong candidate as a gene that affects growth traits that could be exploited in a QC cattle breeding program. More generally, our data provide a new theoretical basis for QC beef breeding and beef quality improvement.展开更多
A key step in the manufacturing of cultured meat is to produce myotubes by induced myogenic differentiation.The development of effective,low-cost,and food-safe components that promote the in vitro differentiation of m...A key step in the manufacturing of cultured meat is to produce myotubes by induced myogenic differentiation.The development of effective,low-cost,and food-safe components that promote the in vitro differentiation of myoblasts is essential for the industrialization of cultured meat.Flavonoids are a class of plant secondary metabolites with various biological activities,but their effects on the regulation of myoblast behaviors are a lack of study.In this study,we selected four representative flavonoids including luteolin,chrysin,apigenin,and genistein,and investigated their effects on porcine myoblasts in the aspects of proliferation,migration,and differentiation.The results showed that four flavonoids all had relatively low cell toxicity but weak ability to promote the proliferation of porcine myoblasts.A positive effect of luteolin was observed in the migration and differentiation of porcine myoblasts,as indicated by improved migration rate and fusion index,as well as upregulated expression of Myogenin and MyHC.Pharmacological inhibition of PI3K activity attenuated the efficacy of luteolin on porcine myoblasts,and further analysis showed that luteolin increased the phosphorylation of PI3K,Akt,and mTOR,indicating the activation of the PI3K/Akt/mTOR signaling pathway.In conclusion,these findings showed that luteolin promoted the migration and differentiation of porcine myoblast via the activation of PI3K/Akt/mTOR signaling,providing biological evidence for its application in cultured meat production.展开更多
Regenerative muscles are required for swallowing and mastication, and are important for functional recovery from diseases involving oral muscular defects. Therefore, we generated three-layer hybrid sheets, similar to ...Regenerative muscles are required for swallowing and mastication, and are important for functional recovery from diseases involving oral muscular defects. Therefore, we generated three-layer hybrid sheets, similar to oral mucosal structures containing submucosal muscles, using rabbit oral mucosa epithelial, mesenchymal, and myoblastic progenitor cells, and examined the structural proteins. Each cell type was obtained from rabbit oral mucosa using enzymatic digestion. Isolated mesenchymal and myoblastic cells were multi-differentiated into osteoblasts, adipocytes, and chondrocytes or myotubes. Isolated epithelial cells were cultured on collagen gels containing isolated mesenchymal cells for 2 weeks, and these epithelial-mesenchymal cell sheets were laminated onto myoblastic cell sheets. The engineered hybrid sheets were multi-stratified in the epithelial and myoblastic layers in a time-dependent manner, expressing intermediate cytoskeletal filament proteins of epithelium and muscle. Hybrid sheets also expressed extracellular matrix basement membrane proteins. Immature cell markers for epithelial and myoblastic cells were observed continuously in hybrid sheet cultures. We established engineered three-dimensional rabbit oral mucosa hybrid sheets containing each immature cell type in vitro.展开更多
基金supported by National Natural Science Foundation of China(Grant No.51872224)Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research,College of Stomatology,Xi’an Jiaotong University(Grant No.2018LHMKFKT004)+1 种基金Special Guidance Funds for the Construction of World-class Universities(disciplines)and Characteristic Development in Central Universities(grant No.PY3A078)the Fundamental Research Funds for the Central Universities(grant No.xzy022019050)。
文摘Complete skeletal muscle repair and regeneration due to severe large injury or disease is still a challenge.Biochemical cues are critical to control myoblast cell function and can be utilized to develop smart biomaterials for skeletal muscle engineering.Citric acid-based biodegradable polymers have received much attention on tissue engineering,however,their regulation on myoblast cell differentiation and mechanism was few investigated.Here,we find that citrate-based polycitrate-polyethylene glycol-polyethylenimine(POCG-PEI600)nanoclusters can significantly enhance the in vitro myoblast proliferation by probably reinforcing the mitochondrial number,promote the myotube formation and full-thickness skeletal muscle regeneration in vivo by activating the myogenic biomarker genes expression of Myod and Mhc.POCG-PEI600 nanoclusters could also promote the phosphorylation of p38 in MAP kinases(MAPK)signaling pathway,which led to the promotion of the myoblast differentiation.The in vivo skeletal muscle loss rat model also confirmed that POCG-PEI600 nanoclusters could significantly improve the angiogenesis,myofibers formation and complete skeletal muscle regeneration.POCG-PEI600 nanocluster could be also biodegraded into small molecules and eliminated in vivo,suggesting their high biocompatibility and biosafety.This study could provide a bioactive biomaterial-based strategy to repair and regenerate skeletal muscle tissue.
文摘旨在了解m6A甲基转移酶样蛋白16(methyltransferase-like protein 16,METTL16)在鸡不同类型肌肉组织中的表达情况及其在鸡骨骼肌功能中的调控作用。本研究利用qRT-PCR技术检测METTL16基因在120日龄广西麻鸡母鸡不同类型肌肉组织中的表达,同时利用siRNA干扰鸡原代成肌细胞中METTL16基因的表达,分析其对成肌细胞增殖、分化和肌纤维形成的影响。结果显示,METTL16基因在鸡的不同类型肌肉组织(胸大肌、胸小肌、缝匠肌、耻坐骨肌内侧肌、耻坐骨肌外侧肌、髂胫外侧肌、腓肠肌内侧肌和背阔肌)中广泛表达,其中,在白肌纤维为主的胸大肌、胸小肌和髂胫外侧肌中表达量相对更高;细胞增殖检测结果显示,干扰METTL16基因表达后,鸡成肌细胞的增殖活力受到了抑制;在成肌细胞分化后干扰METTL16基因表达,鸡成肌细胞RNA m6A甲基化水平呈下降趋势,细胞分化关键基因MyoD表达显著下降(P<0.05),fast-MyHC基因表达显著上升(P<0.001),slow-MyHC基因表达呈下降趋势。综上,METTL16基因在不同类型肌肉中的表达与其肌纤维类型组成相关,METTL16基因可能在鸡成肌细胞增殖和分化中发挥重要作用,并抑制快肌纤维的形成,提示METTL16可能在鸡肌肉功能调控中发挥重要作用。
文摘Sirt1(Sirtuin type 1)是依赖于烟酰胺腺嘌呤二核苷酸(NAD+)的组蛋白脱乙酰酶,为Sirtuins家族成员之一,与细胞增殖、分化、衰老、凋亡和代谢密切相关。目前,有关Sirt1与衰老和代谢的论文已在Science、Nature、Cell等杂志上连续刊出。其中,Sirt1通过抑制PPARγ促进白色脂肪细胞中脂肪动员,并且通过下调肌细胞标志基因表达来抑制成肌细胞分化。提示Sirt1不仅是一个重要的与机体“长寿”有关的因子,而且可能在动物脂肪沉积和肌肉发育中起着关键的调控作用。
文摘Growth and differentiation factor-11 (GDF-11) is a secreted protein that is closely related to myostatin, a known inhibitor of skeletal muscle development. The role of GDF-11 in regulating skeletal muscle growth remains unclear and the pattern of expression during post-natal growth has not been reported. Therefore, we sought to determine the expression of GDF-11 during post-natal growth and its effect on myoblast proliferation and differentiation. We collected gastrocnemius muscles from male and female mice at 2, 3, 4, 6, 12, 20 and 32 weeks of age (n = 6 per sex and age). In addition, gastrocnemius muscles were col- lected from male wild-type and myostatin knockout mice at 4, 6, 12 and 20 weeks of age (n = 6 per age and genotype). RNA was extracted and reverse tran- scribed. Northern analysis identified an expected 4.4 kb mRNA transcript for GDF-11 in gastrocnemius muscles of mice. The concentration of GDF-11 mRNA, as determined by quantitative PCR, was increased in gastrocnemius muscles from 2 to 6 weeks—a period of rapid postnatal muscle growth—and remained higher in male than female mice from 4 to 20 weeks of age (P gastrocnemius muscles of myostatin knockout compared with wild-type mice (P < 0.05), which may suggest a compensatory mecha- nism for the lack of myostatin. In support, recombi- nant GDF-11 inhibited differentiation of C2C12 mur- ine myoblasts and those isolated from myostatin knockout and wild-type mice (P < 0.05). Inhibited dif-ferentiation of C2C12 myoblasts was associated with decreased mRNA expression of early and late mo- lecular markers of differentiation (MyoD, myogenin, IGF-II, desmin and MyHC, P < 0.05). Our results suggest that GDF-11 regulates growth of skeletal muscles by inhibiting myoblast differentiation in an autocrine/paracrine manner and, perhaps, also plays a role in regulating sexually dimorphic growth.
基金supported by the National Natural Science Foundation of China (31272408)Agricultural Science and Technology Innovation Projects of Shaanxi Province (2012NKC01-13)+1 种基金Program of National Beef Cattle Industrial Technology System (CARS-38)National High Technology Research and Development Program of China (2013AA102505)
文摘Our previous work reported a relationship between FOXO1 mutations and growth of Qinchuan(QC) cattle. Here, we performed differential expression analysis of FOXO1 and its association analysis with growth traits in QC cattle. First, we measured the expression of the FOXO1 gene in nine tissues during three developmental stages. The results showed that FOXO1 was abundantly expressed in tissues of calves but was strongly repressed in adulthood, although there was significant transcription in skeletal muscle. FOXO1 expression showed gradual up-regulation during differentiation of primary bovine skeletal muscle cells.We also identified six SNPs of the bovine FOXO1 gene by sequencing DNA pools of samples from 488 individuals, and association analysis indicated that five SNPs were significantly associated with some growth traits in the QC population. We further analyzed four haplotype combinations of the six SNPs and found significant correlation with body length(P<0.01). In conclusion, FOXO1 participates in bovine myocyte differentiation and expression, and may be a strong candidate as a gene that affects growth traits that could be exploited in a QC cattle breeding program. More generally, our data provide a new theoretical basis for QC beef breeding and beef quality improvement.
基金financially supported by the Innovative Research Groups Project of the National Natural Science Foundation of China(32021005)the National Key Research and Development Program of China(2021YFC2101400)+4 种基金the Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study(SN-ZJU-SIAS-0013)the National Natural Science Foundation of China(3210160732)the Natural Science Foundation of Jiangsu Province(BK20210462)the Natural Science Foundation of Jiangsu Province(BK20202002)the Undergraduate Innovation Training Program of China(202110295007Z).
文摘A key step in the manufacturing of cultured meat is to produce myotubes by induced myogenic differentiation.The development of effective,low-cost,and food-safe components that promote the in vitro differentiation of myoblasts is essential for the industrialization of cultured meat.Flavonoids are a class of plant secondary metabolites with various biological activities,but their effects on the regulation of myoblast behaviors are a lack of study.In this study,we selected four representative flavonoids including luteolin,chrysin,apigenin,and genistein,and investigated their effects on porcine myoblasts in the aspects of proliferation,migration,and differentiation.The results showed that four flavonoids all had relatively low cell toxicity but weak ability to promote the proliferation of porcine myoblasts.A positive effect of luteolin was observed in the migration and differentiation of porcine myoblasts,as indicated by improved migration rate and fusion index,as well as upregulated expression of Myogenin and MyHC.Pharmacological inhibition of PI3K activity attenuated the efficacy of luteolin on porcine myoblasts,and further analysis showed that luteolin increased the phosphorylation of PI3K,Akt,and mTOR,indicating the activation of the PI3K/Akt/mTOR signaling pathway.In conclusion,these findings showed that luteolin promoted the migration and differentiation of porcine myoblast via the activation of PI3K/Akt/mTOR signaling,providing biological evidence for its application in cultured meat production.
基金partly supported by an Assistance for Joint Research grant with the Community Program in Life Sciences from the Ministry of Education,Culture,Sports,Science and Technology of Japan
文摘Regenerative muscles are required for swallowing and mastication, and are important for functional recovery from diseases involving oral muscular defects. Therefore, we generated three-layer hybrid sheets, similar to oral mucosal structures containing submucosal muscles, using rabbit oral mucosa epithelial, mesenchymal, and myoblastic progenitor cells, and examined the structural proteins. Each cell type was obtained from rabbit oral mucosa using enzymatic digestion. Isolated mesenchymal and myoblastic cells were multi-differentiated into osteoblasts, adipocytes, and chondrocytes or myotubes. Isolated epithelial cells were cultured on collagen gels containing isolated mesenchymal cells for 2 weeks, and these epithelial-mesenchymal cell sheets were laminated onto myoblastic cell sheets. The engineered hybrid sheets were multi-stratified in the epithelial and myoblastic layers in a time-dependent manner, expressing intermediate cytoskeletal filament proteins of epithelium and muscle. Hybrid sheets also expressed extracellular matrix basement membrane proteins. Immature cell markers for epithelial and myoblastic cells were observed continuously in hybrid sheet cultures. We established engineered three-dimensional rabbit oral mucosa hybrid sheets containing each immature cell type in vitro.
