泥鳅粉对训练小鼠游泳力竭24h后心肌S100A4蛋白表达的影响(1)
摘要：目的 从心肌S100A4蛋白表达变化探讨生泥鳅干预对游泳训练力竭小鼠心肌的作用。方法 24只健康雄性ICR小鼠分为安静组(C)、运动组(T)、泥鳅运动组(N)，T组和N组游泳训练3w后进行力竭运动，24h后取材检测血清CK-MB，HE染色光镜下观察心肌组织，免疫组化法检测心肌P53和S100A4蛋白的表达。结果 ①N组力竭游泳时间显著地长于T组(P<0.01)，血清CK-MB活性显著地低于T组和C组(P<0.05)。②T组的心肌细胞界限模糊、可见心肌纤维断裂和间质水肿，而N组无明显病理表现。③与C组比较，T组心肌P53蛋白的表达显著增高(P﹤0.05)；S100A4蛋白N组显著增高(P﹤0.05)。结论 负荷游泳训练并力竭24 h后小鼠的心肌细胞存在损害，生泥鳅干预能上调S100A4表达和减弱P53表达而降低损害。
关键词：生泥鳅；力竭运动；心肌；S100A4蛋白
Abstract：Objective This study was to observe effects of raw loach on myocardium by expression of S100A4. Methods 24 healthy male ICR mouse were divided into control group(C)， exercise-trained group(T)and exercise + raw loach group (N). The exhaustive exercise time of mice was recorded after three week exercise-trained. Activity of serum MB isoenzyme of creatine kinase (CK-MB) was detected. The myocardium were observed by microscope after HE taining.The expressions of S100A4 protein were detected with immunohistochemistry. Results ①The time of exercise exhaustive were longer than group T(P﹤0.01)， the activity of CK-MM were lower than group T and C(P<0.05) in group N. ②There was not obviously tissue damage in group N. ③Compared with group C， the expression of P53 protein increased significantly in group T(P<0.05) and the expression of S100A4 protein was increased significantly in group N(PP<0.05). Conclusion There was myocardial injury in mice with load swimming training at 24 Hours after exhaustive exercise. But raw loach can reduce the damage of myocardia by increased the expression of S100A4 protein.
, http://www.100md.com
Key words：Loach； Exhaustion exercise；Myocardial； S100A4 protein
泥鳅味道鲜美、氨基酸组成全面，必需氨基酸构成比例符合FAO/WHO的标准[1]，含有大量益于人体的生物活性物质。有研究报道，泥鳅酶解蛋白、冻干粉和中药复合剂有强身保健、抗疲劳和抗自由基保护机体作用[1-3]，而对保护运动机体的效果和机制国内外研究极少。众所周知，力竭运动力竭运动对心肌产生损害。近年研究发现，一种钙结合蛋白S100A4能促血管生成和对心脏损伤起保护作用[4]。本实验生泥鳅粉应用于游泳训练小鼠，观察力竭游泳能力、心肌组织及心肌S100A4蛋白表达变化，探索泥鳅保护运动机体心肌的作用与机制。
1资料与方法
1.1一般资料 健康雄性ICR小鼠，体重(20±2)g，购自湖南斯耐克景达实验动物有限公司，许可证号SCXK(湘)2011-0003。以国家标准啮齿类动物饲料分笼饲养，自由饮食和饮水。室温控制在(22±2)℃，相对湿度40%～55%，光照12h/d。小鼠静养与观察3d。按体重分层随机分为安静对照组(C)、运动训练组(T)、运动训练+泥鳅组(N)，每组8只。
, http://www.100md.com
1.2游泳训练与力竭 运动组小鼠放入深80 cm、直径55 cm桶内游泳训练，水温(30±2)℃、水深40 cm。5d/w，下午进行，共进行3w。第1w不负重30min，第2w尾根部2%负重40min，第3w 3%负重40min。第2w开始，每天上午10点灌胃，量0.3 ml/只，C组和T组灌生理盐水，N组灌生泥鳅溶液。训练结束休息2d，T组和N组尾根部负重5%进行力竭游泳，记录游泳开始至力竭的时间。力竭判断标准：小鼠沉入水中超过10s，且放在平面时呼吸深急，神情疲倦，垂头俯卧，刺激后无反应，无法完成翻正反射。
1.3泥鳅灌胃液制备 将生鲜市场所购活泥鳅放入水中静养2～3d，清洗沥干，加液氮速冻除头和内脏、剁碎，在陶瓷碾砵内碾磨成粉，-80℃保存。参考冻干泥鳅粉的文献有效量[2]，按2.5g/kg量用纯水将泥鳅粉配制成16.67%(w/v)灌胃液。
1.4取材 小鼠力竭游泳24h后，与C组均称取体重和麻醉，摘眼球采血分离血清，-80℃保存，检测血清CK-MB活性。在冰上打开胸腔摘取心脏，放入预冷的冰生理盐水中清洗，滤纸吸干水分并切取左心室心肌约3×3×3 mm置入4℃的4%多聚甲醛中，固定1w进行常规组织脱水、透明、包埋、石蜡切片。, http://www.100md.com(刘祥梅 施孟)
关键词：生泥鳅；力竭运动；心肌；S100A4蛋白
Abstract：Objective This study was to observe effects of raw loach on myocardium by expression of S100A4. Methods 24 healthy male ICR mouse were divided into control group(C)， exercise-trained group(T)and exercise + raw loach group (N). The exhaustive exercise time of mice was recorded after three week exercise-trained. Activity of serum MB isoenzyme of creatine kinase (CK-MB) was detected. The myocardium were observed by microscope after HE taining.The expressions of S100A4 protein were detected with immunohistochemistry. Results ①The time of exercise exhaustive were longer than group T(P﹤0.01)， the activity of CK-MM were lower than group T and C(P<0.05) in group N. ②There was not obviously tissue damage in group N. ③Compared with group C， the expression of P53 protein increased significantly in group T(P<0.05) and the expression of S100A4 protein was increased significantly in group N(PP<0.05). Conclusion There was myocardial injury in mice with load swimming training at 24 Hours after exhaustive exercise. But raw loach can reduce the damage of myocardia by increased the expression of S100A4 protein.
, http://www.100md.com
Key words：Loach； Exhaustion exercise；Myocardial； S100A4 protein
泥鳅味道鲜美、氨基酸组成全面，必需氨基酸构成比例符合FAO/WHO的标准[1]，含有大量益于人体的生物活性物质。有研究报道，泥鳅酶解蛋白、冻干粉和中药复合剂有强身保健、抗疲劳和抗自由基保护机体作用[1-3]，而对保护运动机体的效果和机制国内外研究极少。众所周知，力竭运动力竭运动对心肌产生损害。近年研究发现，一种钙结合蛋白S100A4能促血管生成和对心脏损伤起保护作用[4]。本实验生泥鳅粉应用于游泳训练小鼠，观察力竭游泳能力、心肌组织及心肌S100A4蛋白表达变化，探索泥鳅保护运动机体心肌的作用与机制。
1资料与方法
1.1一般资料 健康雄性ICR小鼠，体重(20±2)g，购自湖南斯耐克景达实验动物有限公司，许可证号SCXK(湘)2011-0003。以国家标准啮齿类动物饲料分笼饲养，自由饮食和饮水。室温控制在(22±2)℃，相对湿度40%～55%，光照12h/d。小鼠静养与观察3d。按体重分层随机分为安静对照组(C)、运动训练组(T)、运动训练+泥鳅组(N)，每组8只。
, http://www.100md.com
1.2游泳训练与力竭 运动组小鼠放入深80 cm、直径55 cm桶内游泳训练，水温(30±2)℃、水深40 cm。5d/w，下午进行，共进行3w。第1w不负重30min，第2w尾根部2%负重40min，第3w 3%负重40min。第2w开始，每天上午10点灌胃，量0.3 ml/只，C组和T组灌生理盐水，N组灌生泥鳅溶液。训练结束休息2d，T组和N组尾根部负重5%进行力竭游泳，记录游泳开始至力竭的时间。力竭判断标准：小鼠沉入水中超过10s，且放在平面时呼吸深急，神情疲倦，垂头俯卧，刺激后无反应，无法完成翻正反射。
1.3泥鳅灌胃液制备 将生鲜市场所购活泥鳅放入水中静养2～3d，清洗沥干，加液氮速冻除头和内脏、剁碎，在陶瓷碾砵内碾磨成粉，-80℃保存。参考冻干泥鳅粉的文献有效量[2]，按2.5g/kg量用纯水将泥鳅粉配制成16.67%(w/v)灌胃液。
1.4取材 小鼠力竭游泳24h后，与C组均称取体重和麻醉，摘眼球采血分离血清，-80℃保存，检测血清CK-MB活性。在冰上打开胸腔摘取心脏，放入预冷的冰生理盐水中清洗，滤纸吸干水分并切取左心室心肌约3×3×3 mm置入4℃的4%多聚甲醛中，固定1w进行常规组织脱水、透明、包埋、石蜡切片。, http://www.100md.com(刘祥梅 施孟)
参见：首页 > 保健版 > 康复中心 > 运动保健 > 游泳