电针干预对大鼠腓肠肌适应性肥大及肌卫星细胞增殖分化的影响  

Effects of Electroacupuncture on Adaptive Hypertrophy of Gastrocnemius and Proliferation and Differentiation of Muscle Satellite Cells in Rats

作　　者：赵丹丹[1,2] 唐成林 黄思琴[1,2] 罗翱 张安宁[1,2] 郭全虎 曹净[1,2] 高睿琦 

机构地区：[1]重庆医科大学中医药学院,重庆400016 [2]中医药防治代谢性疾病重庆市重点实验室,重庆400016

出　　处：《针刺研究》2017年第6期489-495,共7页Acupuncture Research

基　　金：国家自然科学基金(No.81273870)

摘　　要：目的:观察不同时长电针干预对大鼠腓肠肌适应性肥大及腓肠肌中增殖细胞核抗原(PCNA)蛋白表达及配对盒转录因子7(Pax 7)、成肌分化抗原(MyoD 1)、肌细胞生成素(MyoG)、肌球蛋白重链-Ⅰ(Myh 7)、转化生长因子-β1(TGF-β1)、Smad 3mRNA表达的影响,探讨电针诱导骨骼肌适应性肥大的相关机制。方法:SD大鼠随机分为空白对照组、电针2周组、电针4周组和电针8周组,每组6只。除空白对照组,其余3组大鼠予以右侧"足三里"和"环跳"电针干预10min,每日1次,每周6次,分别持续干预2、4和8周。称重后计算各组大鼠腓肠肌湿重比,HE染色后测定腓肠肌肌纤维截面积及直径,免疫荧光标记法观察腓肠肌中PCNA蛋白的表达改变,实时荧光定量PCR法检测腓肠肌中Pax 7、MyoD 1、MyoG、Myh 7、TGF-β1、Smad 3mRNA相对表达量。结果:与空白对照组相比,腓肠肌中PCNA蛋白表达在电针2周组和电针4周组明显增多;大鼠腓肠肌湿重比、肌纤维截面积和直径及腓肠肌Pax 7、MyoD 1、MyoG、Myh 7、TGF-β1mRNA相对表达量随电针干预时间延长呈现先升高后降低的趋势,与空白对照组比较,Pax 7和TGF-β1mRNA相对表达量在干预2周时到达峰值(P<0.01),肌湿重比、肌纤维截面积和直径及MyoD 1、MyoG、Myh 7mRNA相对表达量在干预4周时到达峰值(P<0.01);腓肠肌中Smad 3mRNA相对表达量随电针干预时间延长呈现先下降后上升的趋势,在电针4周组到达低峰(P<0.01)。结论:电针干预"足三里"和"环跳"诱导大鼠腓肠肌产生时序性适应性肥大的改变可能是通过上调腓肠肌中Pax 7、PCNA、TGF-β1、MyoD 1、MyoG、Myh 7的表达,并下调Smad 3的表达,促进肌卫星细胞的增殖、成肌分化和肌管的终末分化,增加电针侧腓肠肌质量、肌纤维面积和直径而实现的。Objective To explore the effects and mechanism of different duration electroacupuncture （EA） on adaptive hypertrophy of gastrocnemius and the expressions of proliferation cell nuclear antigen （PCNA）, paired box transcription factor 7 （Pax 7）, myogenic differentiation antigen （MyoD 1）, myogenin （MyoG）, myosin heavy chain- T （Myh 7）, transforming growth factor-β 1 （TGF-β1）, Smad 3 in rats. Methods Twenty-four SD adult male rats were randomly divided into normal, two-week EA （A 2）, four-week EA （A 4） and eight-week EA （A 8） groups, 6 rats in each group. Except the normal group, EA was applied at right ＂Zusanli＂ （ST 36） and ＂Huantiao＂ （GB 30） for 10 min in the other three groups, separately for 2 weeks, 4 weeks and 8 weeks, once a day, 6 times a week. The ratio of gastrocnemius wet weight was calculated after EA. The cross-sectional area and diameter of muscle fiber were measured after HE staining. The expression of PCNA protein was detected by immunofluorescence. The relative expressions of Pax 7, MyoD 1, MyoG, Myh 7, TGF-β1 and Smad 3 mRNA were tested with real-time PCR. Results The expressions of PCNA protein were obviously higher in the A 2 and A 4 groups than that in the normal group. The expression of genes in the EA intervention groups showed a trend of increasing first and then decreasing except Smad 3 mRNA. The relative expressions of Pax 7 and TGF-β 1 mRNAs reached their peaks in the A 2 group compared with those in the normal group （ P〈0.01）. The ratio of gastrocnemius wet weight, cross-sectional area and diameter of muscle fiber, and the relative expressions of MyoD 1, MyoG, Myh 7 mRNAs reached their peaks in the A 4 group （ P〈0.01 ）, while the relative expression of Smad 3 mRNA showed a trend of decreasing first and then increasing and reached its bottom at the 4th week （P〈0.01）. Conclusion EA at ＂Zusanli＂ （ST 36） and ＂Huantiao＂ （GB 30） may induce adaptive hypertrophy of gastrocnemius in rats by up-regulating

关 键 词：电针 肌卫星细胞 骨骼肌肥大 配对盒转录因子7 成肌分化抗原 

分 类 号：R245.97[医药卫生—针灸推拿学]