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1. 北京中医药大学针灸推拿学院
2. 广东省第二中医院针灸康复科
3. 广东省中医院康复科
4. 北京大学第三医院中医科
纸质出版日期:2016
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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.
LIU Tong, YU Jia-ni, ZOU De-hui, et al. Effect of Electroacupuncture Serum on Proliferation of Cultured Multifidus Muscle Satellite Cells and Expression of Pax-7,MyoD and p-Akt[J]. Acupuncture research, 2016, 41(5): 402-409.
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. DOI: 10.13702/j.1000-0607.2016.05.008.
LIU Tong, YU Jia-ni, ZOU De-hui, et al. Effect of Electroacupuncture Serum on Proliferation of Cultured Multifidus Muscle Satellite Cells and Expression of Pax-7,MyoD and p-Akt[J]. Acupuncture research, 2016, 41(5): 402-409. DOI: 10.13702/j.1000-0607.2016.05.008.
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(P<0.05
P<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(P<0.01
P<0.05)
but not in the EA-BL 40serum+LY 294002 and EA-BL 23serum+LY 294002groups(P>0.05)
suggesting an involvement of PI 3Kin 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-7proteins among the 6serum groups(P>0.05).Conclusion 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 3Ksignaling.
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