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.
School of Acu-moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China; Department of Rehabilitation, Guangdong Hospital of Traditional Chinese Medicine, Guangzhou 510120.
Zhen Ci Yan Jiu. 2016 Oct 25;41(5):402-9.
To observe the effect of electroacupuncture (EA) serum on proliferation of multifidus muscle sa-tellite 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.
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 (400 μL) into the bilateral L-L paravertebral muscles (4 points, 100 μL for each point). EA stimulation was separately applied to bilateral BL 40 and BL 23 for 20 min, once daily, 4 days altogether. Blood samples of the abdominal artery of rats in the above mentioned 4 groups 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 40 serum+LY 294002 (an inhibitor of phosphotidylinsitol-3-kinase, PI 3 K), EA-BL 23 serum and EA-BL 23 serum+LY 294002 groups for ana-lyzing 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.
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 (<0.05, <0.01), 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 (<0.01, <0.05), but not in the EA-BL 40 serum+LY 294002 and EA-BL 23 serum+LY 294002 groups (>0.05), suggesting an involvement of PI 3 K in the proliferation of SCs. No marked differences were found in the proliferation levels between the EA-BL 23 and EA-BL 40 serum groups and in the expression levels of Pax-7 proteins among the 6 serum groups (>0.05).
Both EA-BL 40 and EA-BL 23 serum can promote proliferation of multifidus muscle SCs, which may contribute to the effect of EA intervention in promoting repair of the injured muscle, partially by way of Akt/PI 3 K signaling.
观察电针血清对多裂肌卫星细胞（SCs）增殖及SCs中配对盒转录因子Pax-7、肌源性决定因子（MyoD）和蛋白激酶B（PKB或Akt）蛋白表达的影响，以探讨其促进多裂肌修复的潜在机制。
将32只SD大鼠随机分为对照组、模型组、电针委中（BL 40）组和电针肾俞（BL 23）组。通过向双侧L₁-L₂椎旁肌（4个点，每点100 μL）注射0.5%盐酸布比卡因（400 μL）建立多裂肌损伤（MFMI）模型。分别对双侧BL 40和BL 23进行电针刺激20分钟，每天1次，共4天。分别采集上述4组大鼠腹主动脉血样提取血清，经灭活、过滤后，分别应用于正常血清、模型血清、电针BL 40血清和电针BL 40血清+LY 294002（磷脂酰肌醇-3-激酶抑制剂，PI 3 K）、电针BL 23血清和电针BL 23血清+LY 294002组的杜氏改良 Eagle培养基（DMEM）中培养多裂肌SCs，分别采用细胞计数试剂盒-8（CCK-8）法和5-乙炔基-2'-脱氧尿苷（EdU）法分析电针血清对SCs增殖状态的影响。通过蛋白质免疫印迹法检测培养SCs中Pax-7、MyoD和磷酸化（p）-Akt蛋白的表达，以鉴定SCs。
与正常血清组相比，模型血清组SCs的增殖水平（CCK-8法和EdU法检测）以及MyoD和p-Akt蛋白的表达水平均显著升高（P<0.05，P<(此处原文有误，推测应为)<0.01）；而与模型血清组相比，电针BL 23血清组和电针BL 40血清组SCs的增殖及MyoD和p-Akt蛋白的表达水平进一步显著升高（P<0.01，P<0.05），但电针BL 40血清+LY 294002组和电针BL来23血清+LY 294002组无明显变化（P>0.05），提示PI 3 K参与SCs的增殖。电针BL 23血清组和电针BL 40血清组的增殖水平以及6个血清组之间Pax-7蛋白的表达水平差异均无统计学意义（P>0.05）。
电针BL 40血清和电针BL 23血清均可促进多裂肌SCs增殖，这可能是电针干预促进损伤肌肉修复的作用机制之一，部分是通过Akt/PI 3 K信号通路实现的。
