电针“委中”在大鼠腰多裂肌损伤修复过程中对自噬相关蛋白Beclin1、p62的影响
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摘要
目的观察电针"委中"对大鼠腰多裂肌损伤后肌肉再生修复过程中自噬相关蛋白Beclin1、p62表达的动态变化。方法将72只雄性SD大鼠随机分为空白组、模型组、电针委中组,每组24只。每组再随机分为1天、3天、7天三个时间点。模型组和电针委中组麻醉后于双侧L4-5多裂肌注射0.5%布比卡因溶液制造多裂肌损伤模型,空白组不做处理,造模后电针委中组进行每日一次的电针委中治疗,模型组不进行针刺干预,3组分别于治疗后的1天、3天、7天同步取材。通过免疫组化法和Western Blotting法观察多裂肌中Beclin1、p62表达含量的动态变化。结果 1天、3天、7天模型组的Beclin1表达含量均高于空白组(P<0.01),3天、7天电针委中组的Beclin1表达含量高于模型组(P<0.05);3天、7天模型组p62表达含量低于空白组(P<0.05);1天、3天、7天电针委中组p62表达低于模型组(P<0.05)。结论电针委中穴可促进自噬蛋白Beclin1的表达上调,提高自噬水平,促进p62蛋白的分解,有利于骨骼肌损伤后的修复。
Objective To observe the effect of electroacupuncture( EA) stimulate at"Weizhong"( BL 40) on the expression of autophagy associated protein Beclin1 and P62 in rats with mul-tifidus muscle injury. Methods A total of 72 rats were randomly divided into blank group,model group and EA"Weizhong"( BL 40) group,24 rats in each group. Both sides of multifidus muscle( L4 and L5) in model group and BL40 electroacupunture group were injected with 0. 5% bupivacaine at 1,3,7 day. The rats in BL40 electroacupunture group accepted electroacupunture once day after model was establishment,the rats in model group were catched and fixed at same time. The expression of Beclin1,p62 was measured by immunohistochemical method and western-blot method. Results The expression of Beclin1 of model group was higher than that of blank group at at 1,3,7 day( P < 0. 01). The expression of Beclin1 of BL40 electroacupunture group was higher than that of model group at day 3 and day 7( P < 0. 05). The expression of p62 of model groups was higher than that of blank group at day 3 and day 7( P < 0. 05). Compared with the model group,the expression of p62 in BL40 electroacupunture group was decreased at 1,3,7 day( P < 0. 05). Conclusion EA"Weizhong"( BL40) can increase the expression of autophagy associated protein Beclin1,and can improve autophagy levels and decrease the expression of P62,which is beneficial to skeletal muscle repair.
Objective To observe the effect of electroacupuncture( EA) stimulate at"Weizhong"( BL 40) on the expression of autophagy associated protein Beclin1 and P62 in rats with mul-tifidus muscle injury. Methods A total of 72 rats were randomly divided into blank group,model group and EA"Weizhong"( BL 40) group,24 rats in each group. Both sides of multifidus muscle( L4 and L5) in model group and BL40 electroacupunture group were injected with 0. 5% bupivacaine at 1,3,7 day. The rats in BL40 electroacupunture group accepted electroacupunture once day after model was establishment,the rats in model group were catched and fixed at same time. The expression of Beclin1,p62 was measured by immunohistochemical method and western-blot method. Results The expression of Beclin1 of model group was higher than that of blank group at at 1,3,7 day( P < 0. 01). The expression of Beclin1 of BL40 electroacupunture group was higher than that of model group at day 3 and day 7( P < 0. 05). The expression of p62 of model groups was higher than that of blank group at day 3 and day 7( P < 0. 05). Compared with the model group,the expression of p62 in BL40 electroacupunture group was decreased at 1,3,7 day( P < 0. 05). Conclusion EA"Weizhong"( BL40) can increase the expression of autophagy associated protein Beclin1,and can improve autophagy levels and decrease the expression of P62,which is beneficial to skeletal muscle repair.
引文
[1]赵宏,刘志顺,谢利民,等.《腰痛针灸临床实践指南》解读[J].中国针灸,2015,35(10):1065-1068.
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[9]赵永军,卢健.骨骼肌自噬的调控机制与Sarcopenia的关联及运动的影响[J].中国运动医学杂志,2013,32(10):929-937.
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[13]余维豪,范维铭,蔡虹,等.“腰背委中求”的临床与机理研究[J].中国针灸,1997,(8):503-504.
[14]杨辉.腰痛的中医辩证及针灸治疗[J].中医临床研究,2011,3(20):70-71.
[15]彭园园,刘通,陈玉佩,等.电针“委中”对布比卡因致大鼠腰多裂肌损伤后再生及组织形态的影响[J].中国针灸,2016,36(3):287-294.
[16]刘通,于佳妮,邹德辉,等.电针血清对多裂肌卫星细胞增殖及Pax-7、成肌分化抗原、磷酸化蛋白激酶B表达的影响[J].针刺研究,2016,41(5):402-409.
[17]张雨睛,张丽慧,魏尔清,等.Beclin 1在凋亡和自噬中的调节作用[J].中国生物化学与分子生物学报,2015(4):331-338.
[18]Yue Z,Horton A,Bravin M,et al.A novel protein complex linking the delta 2 glutamate receptor and autophagy:implications for neurodegeneration in lurcher mice.[J].Neuron,2002,35(5):921.
[19]Sun Q,Fan W,Chen K,et al.Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase[J].Proceedings of the National Academy of Sciences of the United States of America,2008,105(49):19211-19216.
[20]屈惠莹,包翠芬,于迪,等.地塞米松诱导的危重病性肌病大鼠骨骼肌p62和泛素的表达[J].中国临床解剖学杂志,2014,32(5):599-603.
[21]秦正红.自噬-生物学与疾病基础卷[M].北京:科学出版社,2015:27-28.
[2]Kay AG.An Extensive Literature Review of the Lumbar Multifidus:Biomechanics[J].Journal of Manual&Manipulative Therapy,2001,9(1):17-39.
[3]Koumantakis GA,Watson PJ,Oldham JA.Trunk muscle stabilization training plus general exercise versus general exercise only:randomized controlled trial of patients with recurrent low back pain.[J].Physical Therapy,2005,85(3):209-225.
[4]Huijbregts PA.Therapeutic Exercise for Lumbopelvic Stabilization:a Motor Control Approach for the Treatment and Prevention of Low Back Pain,ed 2.(Book Review)[J].Physical Therapy,2005(5).
[5]邹德辉,卢宗孝,晏珺,等.电针“委中”对大鼠腰多裂肌损伤后LIF、IL-17表达的影响[J].环球中医药,2017,10(4):436-441.
[6]Taguchi T,Hoheisel U,Mense S.Dorsal horn neurons having input from low back structures in rats[J].Pain,2008,138(1):119-129.
[7]李忠仁.实验针灸学[M].中国中医药出版社,2003:316.
[8]Kaushik S,Cuervo A M.Autophagy as a cell-repair mechanism:activation of chaperone-mediated autophagy during oxidative stress[J].Molecular aspects of medicine,2006,27(5):444-454.
[9]赵永军,卢健.骨骼肌自噬的调控机制与Sarcopenia的关联及运动的影响[J].中国运动医学杂志,2013,32(10):929-937.
[10]Neel,Brian A,Lin,et al.Skeletal muscle autophagy:a new metabolic regulator[J].Trends in Endocrinology&Metabolism Tem,2013,24(12):635.
[11]贾红玲,张学伟,张永臣.数据挖掘技术在针灸治疗腰痛古代文献中的应用[J].针灸临床杂志,2013,29(11):40-41.
[12]许军峰.委中穴不同刺法对中风患者下肢功能障碍的影响[J].上海针灸杂志,2009,28(8):439-441.
[13]余维豪,范维铭,蔡虹,等.“腰背委中求”的临床与机理研究[J].中国针灸,1997,(8):503-504.
[14]杨辉.腰痛的中医辩证及针灸治疗[J].中医临床研究,2011,3(20):70-71.
[15]彭园园,刘通,陈玉佩,等.电针“委中”对布比卡因致大鼠腰多裂肌损伤后再生及组织形态的影响[J].中国针灸,2016,36(3):287-294.
[16]刘通,于佳妮,邹德辉,等.电针血清对多裂肌卫星细胞增殖及Pax-7、成肌分化抗原、磷酸化蛋白激酶B表达的影响[J].针刺研究,2016,41(5):402-409.
[17]张雨睛,张丽慧,魏尔清,等.Beclin 1在凋亡和自噬中的调节作用[J].中国生物化学与分子生物学报,2015(4):331-338.
[18]Yue Z,Horton A,Bravin M,et al.A novel protein complex linking the delta 2 glutamate receptor and autophagy:implications for neurodegeneration in lurcher mice.[J].Neuron,2002,35(5):921.
[19]Sun Q,Fan W,Chen K,et al.Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase[J].Proceedings of the National Academy of Sciences of the United States of America,2008,105(49):19211-19216.
[20]屈惠莹,包翠芬,于迪,等.地塞米松诱导的危重病性肌病大鼠骨骼肌p62和泛素的表达[J].中国临床解剖学杂志,2014,32(5):599-603.
[21]秦正红.自噬-生物学与疾病基础卷[M].北京:科学出版社,2015:27-28.