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  

Effect of Electroacupuncture Serum on Proliferation of Cultured Multifidus Muscle Satellite Cells and Expression of Pax-7,MyoD and p-Akt

作　　者：刘通[1,2] 于佳妮[1,3] 邹德辉[1] 陈玉佩[1] 卢宗孝 晏珺 张莉[1] 霍则军[4] LIU Tong YU Jia-ni ZOU De-hui CHEN Yu-pei LU Zong-xiao YAN Jun ZHANG Li HUO Ze- jun(School of Acu-moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China Department of Acupuncture and Rehabilitation, Guangdong Second Hospital of Traditional Chinese Medicine, Guangzhou 510095 Department of Rehabilitation, Guangdong Hospital of Traditional Chinese Medicine, Guangzhou 510120 Department of Chinese Medicine, Peking University 3rd Hospital, Beijing 100191)

机构地区：[1]北京中医药大学针灸推拿学院,北京100029 [2]广东省第二中医院针灸康复科,广州510095 [3]广东省中医院康复科,广州510120 [4]北京大学第三医院中医科,北京100191

出　　处：《针刺研究》2016年第5期402-409,共8页Acupuncture Research

基　　金：国家自然科学基金项目(No.81574052)

摘　　要：目的:观察电针血清对大鼠多裂肌卫星细胞增殖及Pax-7、成肌分化抗原(MyoD)、磷酸化蛋白激酶B(p-Akt)蛋白表达的影响,从血清学角度探讨电针促进多裂肌损伤修复的部分作用机制。方法:SD大鼠随机分为正常组、模型组、电针委中组、电针肾俞组,每组各8只,采用0.5%布比卡因肌肉注射复制多裂肌损伤模型,各电针组分别选取"委中"穴、"肾俞"穴进行电针治疗,每日1次,每次20min,4d后收集血清。原代培养大鼠腰多裂肌卫星细胞并进行鉴定,随机分为正常血清组、模型血清组、电针委中血清组、电针委中血清+LY 294002组、电针肾俞血清组、电针肾俞血清+LY 294002组,分别以各组血清培养24h,CCK-8、EdU法观察肌卫星细胞的增殖,Western blot法检测肌卫星细胞Pax-7、MyoD、p-Akt蛋白表达。结果:与正常血清组相比,模型血清组多裂肌卫星细胞增殖速度明显加快(P<0.01),两电针血清组又高于模型血清组(P<0.01)。与正常血清组相比,模型血清组MyoD、p-Akt表达明显升高(P<0.05),两电针血清组高于模型血清组(P<0.05,P<0.01)。与电针血清组相比,加LY 294002后,细胞增殖速度及MyoD、p-Akt表达均显著下降(P<0.01,P<0.05)。各组之间Pax-7蛋白表达差异无统计学意义(P>0.05)。结论:电针"委中"血清和电针"肾俞"血清均可通过磷脂酰肌醇3-激酶-蛋白激酶B通路促进肌卫星细胞的增殖,且有促进成肌分化的趋势。Objective To observe the effect of electroacupuncture（EA）serum on proliferation of multifidus muscle satellite cells（SCs）and expression of paired box transcription factor Pax-7,MyoD and protein kinase B（PKB or Akt）proteins of SCs,so as to explore its underlying mechanism in promoting repair of multifidus muscles.Methods Thirty-two SD rats were randomly assigned to control,model,EA-Weizhong（BL 40）and EA-Shenshu（BL 23）groups.The multifidus muscle injury（MFMI）model was established by injection of 0.5% bupivacaine hydrochloride（400μL）into the bilateral L4-L5 paravertebral muscles（4points,100μL for each point）.EA stimulation was separately applied to bilateral BL 40 and BL 23 for 20min,once daily,4days altogether.Blood samples of the abdominal artery of rats in the above mentioned 4groups were separately collected for extracting serum,followed by deactivation and filtration,and then were respectively applied to the Dulbecco＇s Modified Eagle Media（DMEM）culturing each multifidus muscle SCs of the normal serum,model serum,EA-BL 40 serum and EA-BL 40serum＋LY 294002（an inhibitor of phosphotidylinsitol-3-kinase,PI 3K）,EA-BL 23 serum and EA-BL 23serum＋LY 294002 groups for analyzing the impact of EA serum on the proliferation state of SCs by Cell Counting Kit-8（CCK-8）and 5-Ethynyl-2＇-deoxyuridine（EdU）methods,respectively.The expression of Pax-7,MyoD and phosphorylated（p）-Akt proteins of the cultured SCs was detected for characterization of SCs by Western blot.Results Compared with the normal serum group,the proliferation levels（detected by both CCK-8-and EdU）and the expression levels of MyoD and p-Akt proteins of SCs in the model serum group were significantly increased（P0.05,P0.01）,while in comparison with the model serum group,the proliferation and expression levels of MyoD and p-Akt proteins of SCs were further significantly increased in both EA-BL 23 and EA-BL 40 serum groups（P0.01,P0.05）,but not in the EA-BL 40serum＋LY 294002 and EA-BL 23serum＋

关 键 词：电针血清 多裂肌卫星细胞 Pax-7 成肌分化抗原 蛋白激酶B 

分 类 号：R245.97[医药卫生—针灸推拿学]