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重庆医科大学中医药学院,中医药防治代谢性疾病重庆市重点实验室,重庆 410007
△邱玮,重庆医科大学硕士研究生。E-mail:2021110759@stu.cqmu.edu.cn
✉唐成林,教授。E-mail:cytcl996@163.com
收稿:2024-01-24,
网络首发:2024-11-04,
纸质出版:2025-01-12
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邱玮, 唐成林, 廖偲, 等. 基于p38 MAPK信号通路探讨电针对大鼠失神经骨骼肌萎缩的影响[J]. 中国针灸, 2025,45(1):61-70.
QIU Wei, TANG Chenglin, LIAO Cai, et al. Effect of electroacupuncture on denervated skeletal muscle atrophy in rats based on p38 MAPK signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(1): 61-70.
邱玮, 唐成林, 廖偲, 等. 基于p38 MAPK信号通路探讨电针对大鼠失神经骨骼肌萎缩的影响[J]. 中国针灸, 2025,45(1):61-70. DOI: 10.13703/j.0255-2930.20240124-k0002.
QIU Wei, TANG Chenglin, LIAO Cai, et al. Effect of electroacupuncture on denervated skeletal muscle atrophy in rats based on p38 MAPK signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(1): 61-70. DOI: 10.13703/j.0255-2930.20240124-k0002.
目的:
2
观察电针对失神经骨骼肌萎缩大鼠步态、氧化应激、炎性反应及蛋白质降解的影响,探讨电针缓解失神经骨骼肌萎缩的可能机制。
方法:
2
将40只8周龄雄性SD大鼠随机分为假手术组、模型组、电针组、p38 MAPK抑制剂组,每组10只。模型组、电针组、p38 MAPK抑制剂组予手术横断右侧坐骨神经构建大鼠失神经骨骼肌萎缩模型,假手术组仅暴露神经,不离断。造模成功1 d后,电针组大鼠予电针干预,穴取右侧“环跳”“足三里”,选择连续波,频率2 Hz,电流1 mA,每次15 min,每天1次,每周6次;p38 MAPK抑制剂组予SB203580(5 mg/kg)腹腔注射,每天1次,每周6次,共干预3周。干预结束后,采用CatWalk XT 10.6动物步态分析仪评定大鼠步行功能。取材后计算大鼠腓肠肌湿重比;HE染色观察大鼠腓肠肌肌纤维形态与横截面积;ELISA法检测大鼠腓肠肌白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α含量;生化羟胺法检测大鼠腓肠肌超氧化物歧化酶(SOD)、丙二醛(MDA)含量;免疫组化法和Western blot法检测大鼠腓肠肌p38丝裂原活化蛋白激酶(p38 MAPK)、磷酸化(p)-p38 MAPK、肌肉萎缩盒F基因(Atrogin-1)、肌肉环状指基因1(Murf-1)、核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)的表达。
结果:
2
与假手术组比较,模型组大鼠站立持续时间、摆动时间、步行周期增加(
P
<
0.001),最大接触时间的足印面积、打印面积、最大接触时间
的平均强度、平均强度、摆动速度、步周长减少(
P
<
0.001),腓肠肌湿重比和肌纤维横截面积降低(
P
<
0.001),腓肠肌IL-6、IL-1β、TNF-α、MDA含量升高(
P
<
0.001),SOD含量降低(
P
<
0.001),腓肠肌p-p38 MAPK、Atrogin-1、Murf-1阳性表达和蛋白表达升高(
P
<
0.001),Nrf2、HO-1阳性表达和蛋白表达降低(
P
<
0.001);与模型组比较,电针组和p38 MAPK抑制剂组大鼠站立持续时间、摆动时间、步行周期减少(
P
<
0.01),最大接触时间的足印面积、打印面积、最大接触时间的平均强度、平均强度、摆动速度、步周长增加(
P
<
0.01),腓肠肌湿重比和肌纤维横断面积增加(
P
<
0.01,
P
<
0.05),腓肠肌IL-6、IL-1β、TNF-α、MDA含量降低(
P
<
0.05,
P
<
0.01),SOD含量升高(
P
<
0.01,
P
<
0.05),腓肠肌p-p38 MAPK、Atrogin-1、Murf-1阳性表达和蛋白表达降低(
P
<
0.05,
P
<
0.01),Nrf2、HO-1阳性表达和蛋白表达升高(
P
<
0.01,
P
<
0.05)。
结论:
2
电针可能通过介导失神经骨骼肌萎缩大鼠骨骼肌p38 MAPK信号通路,进而抑制氧化应激、炎性反应和蛋白质降解,缓解骨骼肌萎缩。
Objective
2
To assess the impacts of electroacupuncture (EA) on the gait
oxidative stress
inflammatory reaction
and protein degradation in the rats of denervated skeletal muscle atrophy
and explore the potential mechanism of EA for alleviating denervated skeletal muscle atrophy.
Methods
2
Forty male SD rats
8 weeks old
were randomly assigned to a sham-surgery group
a model group
an EA group
and a p38 MAPK inhibitor group
with 10 rats in each group. The right sciatic nerve was transected to establish a rat model of denervated skeletal muscle atrophy in the model group
the EA group and the p38 MAPK inhibitor group. In the sham-surgery group
the nerve was exposed without transection. One day after successful modeling
the rats in the EA group received EA at "Huantiao" (GB30) and "Zusanli" (ST36) on the right side
using a continuous wave with a frequency of 2 Hz and current intensity of 1 mA
for 15 min in each session
EA was delivered once a day
6 times a week. In the p38 MAPK inhibitor group
the rats received the intraperitoneal injection with SB203580 (5 mg/kg)
once a day
6 times a week. The intervention was composed of 3 weeks in each group. After the intervention completion
the CatWalk XT 10.6 animal gait analysis system was used to record the gait parameters of rats. The wet weight ratio of the gastrocnemius muscle was calculated after the sample collected. Using HE staining
the fiber morphology and cross-sectional area of the gastrocnemius muscle were observed; ELISA was employed to measure the content of interleukin (IL)-6
IL-1β
and tumor necrosis factor (TNF)-α in the gastrocnemius muscle; the biochemical hydroxyamine method was adopted to detect the content of superoxide dismutase (SOD) and malondialdehyde (MDA) in the gastrocnemius muscle; with immunohistochemistry and Western blot used
the expression of p38 mitogen-activated protein kinase (p38 MAPK)
phosphorylated (p)-p38 MAPK
muscle atrophy F-box gene (Atrogin-1)
muscle RING finger 1 (Murf-1)
nuclear factor E2-related factor 2 (Nrf2)
and heme oxygenase-1 (HO-1) was detected in the gastrocnemius muscle.
Results
2
Compared to the sham-surgery group
in the model group
the standing duration
the swing time and the step cycle were increased (
P
<
0.001)
the footprint area of the maximum contact time
the print area
the average intensity of the maximum contact time
the average intensity
the swing speed
and the step length were decreased (
P
<
0.001); the wet weight ratio of gastrocnemius muscle and fiber cross-sectional area were reduced (
P
<
0.001); the content of IL-6
IL-1β
TNF-α and MDA in gastrocnemius muscle elevated (
P
<
0.001)
and that of SOD reduced (
P
<
0.001); the positive and protein expression of p-p38 MAPK
Atrogin-1 and Murf-1 elevated (
P
<
0.001) and that of Nrf2 and HO-1 dropped (
P
<
0.001). When compared with t
he model group
in the EA group and the p38 MAPK inhibitor group
the standing duration
the swing time and the step cycle decreased (
P
<
0.01)
the footprint area of the maximum contact time
the print area
the average intensity of the maximum contact time
the average intensity
the swing speed
and the step length increased (
P
<
0.01); the wet weight ratio of gastrocnemius muscle and fiber cross-sectional area were improved (
P
<
0.01
P
<
0.05); the content of IL-6
IL-1β
TNF-α and MDA in gastrocnemius muscle dropped (
P
<
0.05
P
<
0.01)
and that of SOD elevated (
P
<
0.01
P
<
0.05); the positive and protein expression of p-p38 MAPK
Atrogin-1 and Murf-1 dropped (
P
<
0.01
P
<
0.05) and that of Nrf2 and HO-1 increased (
P
<
0.01
P
<
0.05).
Conclusion
2
Electroacupuncture may alleviate skeletal muscle atrophy in denervated skeletal muscle atrophy rats by mediating the p38 MAPK activity
thereby suppressing oxidative stress
inflammatory reaction
and protein degradation.
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