| 刘祥华,罗湘筠,李文倩.针刺激活轴抑制蛋白1、腺苷酸活化蛋白激酶、丝/苏氨酸蛋白激酶信号通路对失神经骨骼肌萎缩大鼠的保护作用[J].安徽医药,2022,26(5):913-917. |
| 针刺激活轴抑制蛋白1、腺苷酸活化蛋白激酶、丝/苏氨酸蛋白激酶信号通路对失神经骨骼肌萎缩大鼠的保护作用 |
| Protective effect of acupuncture on denervated skeletal muscle atrophy rats by activating Axin1/AMPK/AKT signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2022.05.015 |
| 中文关键词: 周围神经损伤 失神经 骨骼肌萎缩 大鼠 丝/苏氨酸蛋白激酶 腺苷酸活化蛋白激酶 轴抑制蛋白1 |
| 英文关键词: Peripheral nerve injuries Denervation Skeletal muscle atrophy Rat AKT AMPK Axin1 |
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| 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(Western blotting)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(cleaved caspase-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)、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)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)、cleaved caspase-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。 |
| 英文摘要: |
| Objective To explore effects of acupuncture on expression levels of serine/threonine protein kinase (AKT), adenine mo-nophosphate 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 de-tect 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 significantly increased in acupuncture group (P<0.05), while apoptosis rate (21.39±3.87)% was significantly decreased (P<0.05), expression of Bcl-2 (0.61±0.05), PCNA (0.72±0.08), p-AKT/AKT (0.82±0.09) and Axin1 (0.62±0.06) was significantly up-regu-lated (P<0.05), and expression of Bax (0.33±0.04), cleaved caspase-3 (0.34±0.04) and p-AMPK/AMPK (0.51±0.03) was significantly down-regulated (P<0.05).Conclusion Acupuncture may delay skeletal muscle atrophy of denervation rats by regulating expression of Axin1/AMPK/AKT. |
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