摘要
目的探讨大鼠骨骼肌去肌腱对碳酸酐酶3(CA3)蛋白表达和磷酸酶活性的影响。方法切断大鼠同侧趾长伸肌或比目鱼肌远心端的肌腱,取术后7、14、28和56 d的两侧(去肌腱组和对照组)趾长伸肌和比目鱼肌肌肉组织,对相邻组织切片分别进行ATPase染色和CA3免疫组化染色,Western blot法分析CA3蛋白表达水平,固相膜上原位酶活性分析CA3的磷酸酶活性。结果去肌腱组比目鱼肌组织形态无显著变化,趾长伸肌的快纤维逐渐肥大。相邻切片染色显示去肌腱组比目鱼肌和趾长伸肌的肌纤维ATPase染色和CA3免疫染色对应关系不变,即CA3免疫染色着色纤维即ATPase染色的型纤维。Western blot分析表明,去肌腱组比目鱼肌和趾长伸肌的CA3蛋白表达水平高于对照组,但CA3的磷酸酶活性降低,且去肌腱组比目鱼肌的CA3的磷酸酶活性下降更为显著。结论去肌腱后骨骼肌CA3蛋白水平逐渐增加而磷酸酶活性降低。反映了骨骼肌运动对CA3影响的复杂性,提示评估运动对骨骼肌影响时需全面考察相关蛋白的表达水平和活性状态。
Aim To investigate the effects of tenotomy on the expression and phosphatase activity of carbonic anhydrase 3(CA3).Methods Tendons of extensor digitorum longus(EDL)and soleus muscle(Sol)were removed from the rats.The operated and contralateral muscles were taken at 7 d,14 d,28 d and 56 d after operation.ATPase staining and CA3 immunohistochemical staining were performed on adjacent sections.Western blot was used to analyze the expression of CA3,and phosphate activity analysis on the nitrocellulose filter following electrophoresisin was applied for CA3 phosphatase.Results There was no significant change in the morphology of the Sol after tenotomy,but the fast fibers in the operated EDL became hypertrophied gradually.Adjacent slice staining showed that the relationship between ATPase staining and CA3 immunostaining of Sol and EDL after tenotomy was unchanged,that is,CA3 immunostaining colored fiber was identified as type I fiber by ATPase staining.Western blot analysis showed that the expression of CA3 in operated Sol and EDL was higher than that in the control side.Contrarily,the CA3 phosphatase activity of the former was lower than that of the latter,especially for the operated Sol.Conclusion After tenotomy,the expression of CA3 gradually increased and the phosphatase activity decreased.This reflects the complexity of the effect of movement on CA3,and also suggests that it is necessary to comprehensively examine the expression and activity of proteins when evaluating the effect of the movement on skeletal muscle.
作者
赵艳玲
吴亦影
顾建帮
刘珂
边伟红
黄河
ZHAO Yan-ling;WU Yi-ying;GU Jian-bang;LIU Ke;BIAN Wei-hong;HUANG He(Department of Geriatrics,Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University,Shanghai 200080;Department of Neurology,Shanghai Tenth People's Hospital Chongming Branch,Shanghai 202157;Department of Neurology,Shanghai Tenth People's Hospital Affiliated to Tongji University,Shanghai 200072,China)
出处
《中国临床神经科学》
2021年第3期257-264,共8页
Chinese Journal of Clinical Neurosciences
基金
上海市崇明区“可持续发展科技创新行动计划”(编号:CKY2020-27)。
关键词
骨骼肌
大鼠
去肌腱
比目鱼肌
趾长伸肌
碳酸酐酶3
磷酸酶活性
skeletal muscle
rat
tenotomy
extensor digitorum longus
soleus
carbonic anhydrase3
phosphatase activity
