目的:通过肌肉注射异体脂肪间充质干细胞(ASCs),观察其对一次性离心运动后不同时间点大鼠骨骼肌调节因子的影响,探讨ASCs注射对离心运动后骨骼肌损伤修复的机制。方法:8周龄雄性SD大鼠一次性离心运动后,在左腿腓肠肌注射生理盐水(PBS),...目的:通过肌肉注射异体脂肪间充质干细胞(ASCs),观察其对一次性离心运动后不同时间点大鼠骨骼肌调节因子的影响,探讨ASCs注射对离心运动后骨骼肌损伤修复的机制。方法:8周龄雄性SD大鼠一次性离心运动后,在左腿腓肠肌注射生理盐水(PBS),右腿腓肠肌注射ASCs,然后,随机分为运动后1天组(D1)、运动后3天组(D3)、运动后7天组(W1)和运动后14天组(W2)。透射电镜观察骨骼肌的超微结构变化;测定血清IGF1、GDF8的含量及骨骼肌调节基因的表达。结果:与PBS相比,ASCs注射明显促进肌纤维的修复。与D1组比较,W1组血清IGF1水平显著降低(P<0.05),GDF8水平显著降低(P<0.05),W2组血清GDF8水平显著升高(P<0.05)。与PBS组相比,ASCs组IGF1 m RNA在W2显著升高(P<0.05);GDF8m RNA在D3显著降低(P<0.05),在W2极显著降低(P<0.01);Atrogin1 m RNA在D3显著降低(P<0.05),在W1时间点极显著降低(P<0.01)。PBS组IGF1 m RNA与血清IGF1含量、GDF8m RNA与血清GDF8含量均显著性正相关(P<0.05);骨骼肌总的PCNA m RNA与总的IGF1m RNA显著性正相关(P<0.05)。结论:离心运动后,肌注异体ASCs可以增加骨骼肌IGF1m RNA的表达,降低GDF8和Atrogin1的m RNA表达。肌注异体ASCs可能不影响细胞因子IGF1和GDF8的分泌,而是通过影响局部基因表达来改善离心运动后肌肉的再生修复。展开更多
The Bakerwal breed of goat in Kashmir valley is a good meat purpose breed of goat. That attains an appreciable body weight under a low input production system. As these goats are mainly reared by Gujjar and Bakarwal t...The Bakerwal breed of goat in Kashmir valley is a good meat purpose breed of goat. That attains an appreciable body weight under a low input production system. As these goats are mainly reared by Gujjar and Bakarwal tribes of the J & K state, so they usually are fed with the weeds, herbs, shrubs and grasses of pastures that will otherwise go waste. These goats constitute the main livestock wealth. With the good productive and reproductive potential, it makes these animals an important animal protein source for developing countries like India. The myostatin gene (GDF8) is important in the physiology of stock animals because its product produces a direct effect on muscle development and consequently also on meat production. The myostatin sequence is known in several mammalian species and shows a high degree of amino acid sequence conservation, although several alterations in the intron and exon regions have been identified. The objective of our work is to characterize the myostatin coding regions using gene sequencing and polymerase chain reaction methods of Capra hircus (Bakerwal breed) and to compare them with the Ovis aries and other livestock species of animal, looking for variations in nucleotide and protein sequences. As mutations in the myostatin gene can inactivate its expression and result in a non-functional protein, which leads to increase in muscle growth in many species. In this way, we are able to identify 3 alterations on the presumed myostatin protein sequence as compared to non double-muscled ovine sequences.展开更多
Growth and Differentiation Factor-8(GDF-8) is a new member of TGF-β super-family.It has been shown that GDF-8 is specifically expressed in skeleton muscle in mouse and its function is to inhibit the growth of muscle ...Growth and Differentiation Factor-8(GDF-8) is a new member of TGF-β super-family.It has been shown that GDF-8 is specifically expressed in skeleton muscle in mouse and its function is to inhibit the growth of muscle cell,so it is named as Myostatin.Here,we amplified 3′half-length GDF-8 cDNA from chicken skeleton muscle by RT-PCR,and cloned it into the prokaryotic expression vector pTrcHisB,which was then transformed into E.coli Top10 cells.The recombinant 6×His-GDF-8 fusion protein expressed in the Top10 cells was purified by Ni+-Affinity Chromatography for future study.展开更多
研究不同pH值下,口服靶向FOXO1与GDF-8基因反义RNA寡核苷酸药物对药效的影响。化学合成靶向FOXO1与GDF-8基因的有效反义RNA寡核苷酸片段,调节药物PH值,通过灌胃方式给药,给药20 d后处死小鼠,进行体重,腿部肌肉增长情况分析。提取腿部肌...研究不同pH值下,口服靶向FOXO1与GDF-8基因反义RNA寡核苷酸药物对药效的影响。化学合成靶向FOXO1与GDF-8基因的有效反义RNA寡核苷酸片段,调节药物PH值,通过灌胃方式给药,给药20 d后处死小鼠,进行体重,腿部肌肉增长情况分析。提取腿部肌肉组织总RNA,用real time PCR检测FOXO1与GDF-8基因的表达。结果表明,服用RNA oligos的试验组小鼠腿部肌肉重量均比对照组肌肉重量增长快。其中pH为7.0的RNA oligos的效果比pH为5.0的效果好,pH9.0的RNAoligos的作用效果最弱。与小鼠体重变化结果一致,real time PCR检测实验组FOXO1与GDF-8基因转录水平较对照组均有明显下降,其中服用pH为7.0的RNA oligos的下降最多。因此,口服靶向FOXO1与GDF-8基因的反义RNA寡核苷酸药物的最适pH值应为中性偏酸。展开更多
文摘目的:通过肌肉注射异体脂肪间充质干细胞(ASCs),观察其对一次性离心运动后不同时间点大鼠骨骼肌调节因子的影响,探讨ASCs注射对离心运动后骨骼肌损伤修复的机制。方法:8周龄雄性SD大鼠一次性离心运动后,在左腿腓肠肌注射生理盐水(PBS),右腿腓肠肌注射ASCs,然后,随机分为运动后1天组(D1)、运动后3天组(D3)、运动后7天组(W1)和运动后14天组(W2)。透射电镜观察骨骼肌的超微结构变化;测定血清IGF1、GDF8的含量及骨骼肌调节基因的表达。结果:与PBS相比,ASCs注射明显促进肌纤维的修复。与D1组比较,W1组血清IGF1水平显著降低(P<0.05),GDF8水平显著降低(P<0.05),W2组血清GDF8水平显著升高(P<0.05)。与PBS组相比,ASCs组IGF1 m RNA在W2显著升高(P<0.05);GDF8m RNA在D3显著降低(P<0.05),在W2极显著降低(P<0.01);Atrogin1 m RNA在D3显著降低(P<0.05),在W1时间点极显著降低(P<0.01)。PBS组IGF1 m RNA与血清IGF1含量、GDF8m RNA与血清GDF8含量均显著性正相关(P<0.05);骨骼肌总的PCNA m RNA与总的IGF1m RNA显著性正相关(P<0.05)。结论:离心运动后,肌注异体ASCs可以增加骨骼肌IGF1m RNA的表达,降低GDF8和Atrogin1的m RNA表达。肌注异体ASCs可能不影响细胞因子IGF1和GDF8的分泌,而是通过影响局部基因表达来改善离心运动后肌肉的再生修复。
文摘The Bakerwal breed of goat in Kashmir valley is a good meat purpose breed of goat. That attains an appreciable body weight under a low input production system. As these goats are mainly reared by Gujjar and Bakarwal tribes of the J & K state, so they usually are fed with the weeds, herbs, shrubs and grasses of pastures that will otherwise go waste. These goats constitute the main livestock wealth. With the good productive and reproductive potential, it makes these animals an important animal protein source for developing countries like India. The myostatin gene (GDF8) is important in the physiology of stock animals because its product produces a direct effect on muscle development and consequently also on meat production. The myostatin sequence is known in several mammalian species and shows a high degree of amino acid sequence conservation, although several alterations in the intron and exon regions have been identified. The objective of our work is to characterize the myostatin coding regions using gene sequencing and polymerase chain reaction methods of Capra hircus (Bakerwal breed) and to compare them with the Ovis aries and other livestock species of animal, looking for variations in nucleotide and protein sequences. As mutations in the myostatin gene can inactivate its expression and result in a non-functional protein, which leads to increase in muscle growth in many species. In this way, we are able to identify 3 alterations on the presumed myostatin protein sequence as compared to non double-muscled ovine sequences.
文摘Growth and Differentiation Factor-8(GDF-8) is a new member of TGF-β super-family.It has been shown that GDF-8 is specifically expressed in skeleton muscle in mouse and its function is to inhibit the growth of muscle cell,so it is named as Myostatin.Here,we amplified 3′half-length GDF-8 cDNA from chicken skeleton muscle by RT-PCR,and cloned it into the prokaryotic expression vector pTrcHisB,which was then transformed into E.coli Top10 cells.The recombinant 6×His-GDF-8 fusion protein expressed in the Top10 cells was purified by Ni+-Affinity Chromatography for future study.
文摘研究不同pH值下,口服靶向FOXO1与GDF-8基因反义RNA寡核苷酸药物对药效的影响。化学合成靶向FOXO1与GDF-8基因的有效反义RNA寡核苷酸片段,调节药物PH值,通过灌胃方式给药,给药20 d后处死小鼠,进行体重,腿部肌肉增长情况分析。提取腿部肌肉组织总RNA,用real time PCR检测FOXO1与GDF-8基因的表达。结果表明,服用RNA oligos的试验组小鼠腿部肌肉重量均比对照组肌肉重量增长快。其中pH为7.0的RNA oligos的效果比pH为5.0的效果好,pH9.0的RNAoligos的作用效果最弱。与小鼠体重变化结果一致,real time PCR检测实验组FOXO1与GDF-8基因转录水平较对照组均有明显下降,其中服用pH为7.0的RNA oligos的下降最多。因此,口服靶向FOXO1与GDF-8基因的反义RNA寡核苷酸药物的最适pH值应为中性偏酸。
