电针对大鼠失神经支配骨骼肌萎缩及IGF-1/PI3K/AKT表达的影响  

Effect of electroacupuncture on denervated skeletal muscle atrophy and expression of IGF-1/PI3K/AKT in rats

作　　者：陈玄[1] 叶笑然[1] 黄晓卿[1] CHEN Xuan;YE Xiaoran;HUANG Xiaoqing(Fujian Research Institution of TCM,Fuzhou 350003,China)

机构地区：[1]福建省中医药研究院,福州350003

出　　处：《中国针灸》2018年第12期1311-1317,共7页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金青年项目:81503657;国家自然科学基金面上项目:81373733;福建省属公益类科研院所基本科研专项:2016 R 1033-6

摘　　要：目的:观察电针对失神经支配骨骼肌萎缩及IGF-1/PI3K/AKT信号通路表达的影响,探讨电针对失神经肌肉萎缩的可能作用机制。方法:36只雄性SD大鼠随机分为对照组、模型组和电针组。模型组和电针组大鼠切断左后肢坐骨神经制备失神经肌萎缩模型。电针组予电针治疗,穴取"足三里"和"上巨虚",疏密波,2 Hz/33 Hz,1.5～2 mA,每次30 min,每日1次,每5次间隔2 d,共20次。对照组与模型组每日同样鼠笼固定,但不电针。疗程结束后观察各组坐骨神经功能指数,称量腓肠肌肌湿重,HE染色测量患/健侧肌纤维横截面积比和肌细胞直径比,TUNEL和免疫荧光双染色的方法评价肌细胞凋亡和增殖分化情况。荧光定量PCR法检测胰岛素样生长因子-1(insulin-like growth factor-1,Igf-1)和蛋白激酶B(protein kinase B,PKB,又被称作Akt)的mRNA表达,免疫蛋白印迹法检测局部肌肉Akt总蛋白及其磷酸化形式p-AKT(Ser473)和p-AKT(Thr308)的表达量。结果:与对照组相比,模型组大鼠的坐骨神经功能指数,患侧/健侧腓肠肌湿重比、肌纤维横截面积比和肌细胞直径比均明显下降,肌细胞凋亡指数明显升高(均P<0.01);与模型组相比,电针组上述指标均得到改善(均P<0.05),与肌卫星细胞增殖分化相关的PAX3和PAX7表达明显增高(均P<0.01),p-AKT(Ser473)蛋白水平增加(P<0.05)。结论:电针可改善失神经肌肉的萎缩情况,减少肌细胞凋亡,促进肌卫星细胞增殖分化,其作用机制可能与提高p-AKT(Ser473)水平、激活AKT信号通路有关。Objective To observe the effects of electroacupuncture (EA) on denervated skeletal muscle atrophy and expression of IGF-1/PI3K/AKT signaling pathway in rats, and to explore the possible mechanisms of it. Methods Thirty-six male SD rats were randomly divided into a control group, a model group and an EA group. Denervated muscle atrophy model was made by cutting off the left hindlimb sciatic nerve in the model group and the EA group. In the EA group, EA, dense-disperse wave, 2 Hz/33 Hz and 1.5-2 mA, was connected at “Zusanli” (ST 36) and “Shangjuxu” (ST 37). The treatment was given for 30 min every time, once a day and a total of 20 times were needed, there was 2 days interval every 5 times. The rats in the control group and model group were fixed in the same cage every day but not applied EA. Sciatic nerve function index (SFI) and ratio of muscle wet weight (RWW) were measured after treatment. HE staining was used to determine the ratio of cross-sectional area (RCA) and the ratio of fiber diameter (RFD). TUNEL and double immunofluorescence staining were used to evaluate the myocyte apoptosis, proliferation and differentiation. The mRNA expressions of IGF-1 and AKT were detected by real-time PCR. The total protein of AKT, p-AKT (Ser473) and p-AKT (Thr308) were detected by western blot. Results Compared with the control group, SFI, RWW, RCA and RFD of the model group were significantly decreased and apoptotic index of myocytes were significantly increased (all P<0.01). Compared with the model group, the above indexes of the EA group were improved (all P<0.05), and the expressions of PAX3and PAX7 related to proliferation and differentiation of muscle satellite cells were significantly increased (both P<0.01), p-AKT (Ser473) protein level was increased (P<0.05). Conclusion EA can improve the atrophy of denervated muscles, reduce myocyte apoptosis, and promote the proliferation and differentiation of muscle satellite cells. The mechanism may be related to the increase of p-AKT (Ser473) level and activation of A

关 键 词：失神经 肌萎缩 电针 穴 足三里 穴 上巨虚 蛋白激酶B(AKT) 

分 类 号：R245.97[医药卫生—针灸推拿学]