摘要
目的研究针刺对失神经骨骼肌萎缩大鼠骨骼肌细胞丝/苏氨酸蛋白激酶(AKT)、腺苷酸活化蛋白激酶(AMPK)、轴抑制蛋白1(Axin1)表达水平的影响。方法将48只SD大鼠采用简单随机分组分为空白对照组、假手术组、模型组和针刺组各12只。模型组和针刺组大鼠采用手术切断坐骨神经制备失神经骨骼肌萎缩大鼠模型,假手术组只暴露坐骨神经,空白对照组不做任何处理。针刺组在造模后进行电针治疗(“足三里”和“承山”穴),每次10 min,连续治疗3周。治疗后,检测各组大鼠绯肠肌组织的湿重比、肌纤维横截面积及肌细胞直径比;HE染色检测各组大鼠绯肠肌损伤;TUNEL染色检测各组大鼠绯肠肌细胞凋亡;蛋白质印迹法(Westernblotting)检测p-AKT、AKT、p-AMPK、AMPK、Axin1、B细胞淋巴瘤2(Bcl-2)、Bcl相关X蛋白(Bax)、活化胱天蛋白酶(cleavedcaspase-3)和增殖细胞核抗原(PCNA)蛋白的表达。结果与空白对照组和假手术组相比,模型组大鼠腓肠肌湿重比(0.38±0.06)、肌纤维截面积比(0.52±0.12)和肌纤维直径比(0.53±0.16)均明显下降(P<0.05),细胞凋亡率(30.85±5.74)%明显升高(P<0.05),Bcl-2(0.40±0.03)、Bax(0.53±0.05)、cleavedcaspase-3(0.58±0.06)、PCNA(0.44±0.05)、p-AKT/AKT(0.54±0.06)、p-AMPK/AMPK(0.73±0.04)和Axin1(0.41±0.05)的表达明显上调(P<0.05);与模型组相比,针刺组大鼠腓肠肌湿重比(0.52±0.07)、肌纤维截面积比(0.64±0.11)和肌纤维直径比(0.66±0.15)均明显增加(P<0.05),细胞凋亡率(21.39±3.87)%明显下降(P<0.05),Bcl-2(0.61±0.05)、PCNA(0.72±0.08)、p-AKT/AKT(0.82±0.09)和Axin1(0.62±0.06)的表达明显上调(P<0.05),Bax(0.33±0.04)、cleavedcaspase-3(0.34±0.04)和p-AMPK/AMPK(0.51±0.03)的表达明显下调(P<0.05)。结论针刺可能通过调控Axin1/AMPK/AKT的表达,延缓失神经大鼠骨骼肌的萎缩。
Objective To explore effects of acupuncture on expression levels of serine/threonine protein kinase(AKT),adenine monophosphate activated protein kinase(AMPK)and Axis inhibition protein 1(Axin1)in skeletal muscle cells of rats with denervation skeletal muscle atrophy.Methods Forty-eight SD rats were randomly assigned into blank group,sham operation group,model group and acupuncture group,12 cases in each group.In model group and acupuncture group,sciatic nerve was surgically cut to prepare rats models of denervation skeletal muscle atrophy.Only the sciatic nerve was exposed in sham group and nothing was done in blank group.In acupuncture group,electroacupuncture(Zusanli point and Bear Hill acupuncture point)was conducted after modeling,10 min/once.They were continuously treated for 3 weeks.After treatment,wet to weight ratio of gastrocnemius muscle tissue,ratio of muscle fiber cross-sectional area to myocyte diameter in each group were detected.The gastrocnemius muscle injury in each group was detected by HE staining.The apoptosis of gastrocnemius muscle in each group was detected by TUNEL staining.Western Blot was performed to detect expression of p-AKT,AKT,p-AMPK,AMPK,Axin1,B cell lymphoma 2(Bcl-2),Bcl-associated X protein(Bax),cleaved caspase-3 and proliferating cell nuclear antigen(PCNA)proteins.Results Compared with blank group and sham operation group,wet to weight ratio of gastrocnemius muscle tissue(0.38±0.06),ratio of muscle fiber cross-sectional area(0.52±0.12)and muscle fiber diameter(0.53±0.16)were significantly decreased in model group(P<0.05),while apoptosis rate(30.85±5.74)%was significantly increased(P<0.05),and expressions of Bcl-2(0.40±0.03),Bax(0.53±0.05),cleaved caspase-3(0.58±0.06),PCNA(0.44±0.05),p-AKT/AKT(0.54±0.06),p-AMPK/AMPK(0.73±0.04)and Axin1(0.41±0.05)were significantly up-regulated(P<0.05).Compared with model group,wet to weight ratio of gastrocnemius muscle tissue(0.52±0.07),ratio of muscle fiber cross-sectional area(0.64±0.11)and muscle fiber diameter(0.66±0.15)were si
作者
刘祥华
罗湘筠
李文倩
LIU Xianghua;LUO Xiangyun;LI Wenqian(Department of Acupuncture and Moxibustion,the First Affiliated Hospital of Hunan College of Traditional Chinese Medicine(Hunan Provincial Hospital of Traditional Chinese Medicine),Zhuzhou,Hunan 412000,China)
出处
《安徽医药》
CAS
2022年第5期913-917,I0001,共6页
Anhui Medical and Pharmaceutical Journal
