针刺对膝骨关节炎模型大鼠股四头肌卫星细胞分化和凋亡的影响及其机制

doi:10.13481/j.1671-587X.20250604

Abstract

Abstract:

Objective To discuss the effect of acupuncture on the differentiation and apoptosis of quadriceps muscle satellite cells in model rats with knee osteoarthritis（KOA）， and to clarify its related mechanism. Methods A total of 40 SPF-grade rats were selected and randomly divided into control group， model group， celecoxib group， and acupuncture group， with 10 rats in each group. The rats in control group only underwent joint cavity incision followed by suturing， while the rats in model group， celecoxib group， and acupuncture group were used to replicate the KOA models. The maximum circumference of the femoral segment of the affected limb， rat body mass， and quadriceps wet weight of the rats in various groups were measured； the quadriceps wet weight maintenance rate and quadriceps wet weight/body mass ratio of the rats in various groups were calculated. HE staining was used to observe the pathomorphology of articular cartilage and quadriceps muscle tissue of the rats in various groups； terminal deoxynucleotidyl transferase （TdT）-mediated dUTP nick end labeling （TUNEL） method was used to detect the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in various groups； immunofluorescence method was used to detect the protein expression levels of interleukin-6 （IL-6）， Janus kinase （JAK）， and signal transducer and activator of transcription 3 （STAT3） in quadriceps muscle tissue of the rats in various groups； Western blotting method was used to detect the expression levels of IL-6/JAK/STAT3 signaling pathway proteins， and muscle satellite cells， and apoptosis-related proteins in quadriceps muscle tissue of the rats in various groups. Results Compared with control group， the affected hind limb circumference， quadriceps wet weight， wet weight maintenance rate， and wet weight/body mass ratio of the rats in model group were significantly decreased （P<0.05）； compared with model group， the affected hind limb circumference， quadriceps wet weight， wet weight maintenance rate， and wet weight/body mass ratio of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）； compared with celecoxib group， the affected hind limb circumference， quadriceps wet weight， wet weight maintenance rate， and wet weight/body mass ratio of the rats in acupuncture group were significantly increased （P<0.05）. The HE staining results showed that the knee articular cartilage of the rats in control group remained intact， chondrocytes were aggregated and horizontally arranged with smooth edges， and quadriceps muscle cells were long cylindrical， orderly arranged， and regular in shape； in model group， the knee articular cartilage was thinner with rough edges， reduced number of cartilage layers， and disordered arrangement， and the quadriceps muscle fibers were disorganized， with some muscle fiber dissolution and muscle cell membrane damage， accompanied by muscle fiber fragments and a large amount of inflammatory exudate； in celecoxib group， the morphology of knee articular cartilage was generally normal， occasionally with irregular cartilage arrangement and reduced thickness， sporadically visible necrotic chondrocytes， quadriceps muscle fibers and sarcolemma were relatively intact， new muscle fibers appeared， some muscle fiber edges were blurred， accompanied by a small amount of cell debris and mild inflammatory infiltration； in acupuncture group， the knee articular cartilage structure remained intact with smooth edges， occasionally rough edges， and chondrocytes were aggregated and orderly arranged. The TUNEL assay results showed that compared with control group， the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in model group were significantly increased （P<0.05）； compared with model group， the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly decreased （P<0.05）； compared with celecoxib group， the apoptosis index in articular cartilage and quadriceps muscle tissue of the rats in acupuncture group were significantly decreased （P<0.05）. The immunofluorescence assay results showed that compared with control group， the expression levels of IL-6， JAK， and STAT3 proteins in quadriceps muscle tissue of the rats in model group were significantly decreased （P<0.05）； compared with model group， the expression levels of IL-6， JAK， and STAT3 proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）； compared with celecoxib group， the expression levels of IL-6， JAK， and STAT3 proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of IL-6， JAK， STAT3， paired box transcription factor 7 （Pax7）， Desmin， Myosin， and Myogenin proteins in quadriceps muscle tissue of the rats in model group were significantly decreased （P<0.05）； compared with model group， the expression levels of IL-6， JAK， STAT3， Pax7， Desmin， Myosin， and Myogenin proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）； compared with celecoxib group， the expression levels of IL-6， JAK， STAT3， Pax7， Desmin， Myosin， and Myogenin proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased （P<0.05）. Compared with control group， the expression levels of B-cell lymphoma 2 （Bcl-2）， B-cell lymphoma-xl （Bcl-xl）， and myeloid cell leukemia 1 （MCL1） proteins in quadriceps muscle tissue in model group were significantly decreased （P<0.05）， and the expression levels of Bcl-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase-3 （Caspase-3） proteins were significantly increased （P<0.05）； compared with model group， the expression levels of Bcl-2， Bcl-xl， and MCL1 proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）， and the expression levels of Bax and Caspase-3 proteins were significantly decreased （P<0.05）； compared with celecoxib group， the expression levels of Bcl-2， Bcl-xl， and MCL1 proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased （P<0.05）， and the expression levels of Bax and Caspase-3 proteins were significantly decreased （P<0.05）. Conclusion Acupuncture can promote the differentiation of quadriceps muscle satellite cells and inhibit muscle cell apoptosis in the model rats with KOA， and the mechanism may be related to the up-regulation of expressions of IL-6， JAK， and STAT3 proteins in the quadriceps muscle tissue.

Key words: Acupuncture, Interleukin-6/Janus kinase/signal transducer and activator of transcription 3 signaling pathway, Muscle satellite cells, Knce osteoarthritis model, Quadriceps muscle

CLC Number:

R684.3

Qu ZHENG,Baoqiang DONG,Xingxing LIN,Xuefeng GUAN,Yu ZHANG,Chaojie WANG,Yiyan Han. Effect of acupuncture on differentiation and apoptosis of quadriceps muscle satellite cells in knee osteoarthritis model rats and its mechanism[J].Journal of Jilin University(Medicine Edition), 2025, 51(6): 1475-1486.

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References 24

[1]	ABRAMOFF B， CALDERA F E. Osteoarthritis［J］. Med Clin N Am， 2020， 104（2）： 293-311.
[2]	谢文慧，张卓莉. 难治性类风湿关节炎的治疗策略［J］. 中国实用内科杂志， 2024， 44（12）： 1006-1010.
[3]	中国医师协会风湿免疫科医师分会骨关节炎学组. 中国膝骨关节炎临床药物治疗专家共识（2023）［J］. 中华内科杂志， 2024， 63（6）： 560-578.
[4]	中华中医药学会. 膝骨关节炎中西医结合诊疗指南（2023年版）［J］. 中医正骨， 2023， 35（6）：1-10.
[5]	郑曲，林星星，张宇，等. 基于 Piezo1/YAP/Caspase3轴探讨针刺保护膝骨性关节炎模型大鼠股四头肌细胞的机制研究［J］. 世界科学技术-中医药现代化， 2025， 27（8）： 2274-2283.
[6]	YANG X F， XUE P P， CHEN H R， et al. Denervation drives skeletal muscle atrophy and induces mitochondrial dysfunction， mitophagy and apoptosis via miR-142a-5p/MFN1 axis［J］. Theranostics， 2020， 10（3）： 1415-1432.
[7]	李嫣晓，陈红霞，程梦蝶，等.铁死亡与心肌纤维化相关信号通路的研究进展［J］. 郑州大学学报（医学版）， 2024， 59（2）： 195-201.
[8]	LIU X X， LI X H， ZHOU J T.Experimental study on replicating knee osteoarthritis by modified Hulth’s modeling method［J］. Zhongguo Zhong Xi Yi Jie He Za Zhi， 2005， 25（12）： 1104-1108.
[9]	薛立功. 经筋理论研究对经络学说形成的启迪意义［A］. //中国针灸学会经筋诊治专业委员会成立大会暨首届中华经筋医学论坛论文集［C］. 2009： 1-7.
[10]	APURBA G， SUDIP B. Biomonitoring the skeletal muscle metabolic dysfunction in knee osteoarthritis in older adults： Is Jumpstart Nutrition^® Supplementation effective［J］. Caspian J Intern Med， 2023， 14（4）： 590-606.
[11]	中医康复临床实践指南·膝骨关节炎制定工作组，王尚全，朱立国，等. 中医康复临床实践指南·膝骨关节炎［J］. 康复学报， 2020， 30（3）： 177-182
[12]	林星星，董宝强，王树东，等. 基于筋膜的经筋基础研究：述评与展望［J］. 中国针灸， 2023， 43（11）： 1338-1342.
[13]	罗序国，熊明洁，江睿. 基于“宗筋主束骨而利机关”理论的中医手法治疗膝骨关节炎的临床观察［J］. 中国医学创新， 2022， 19（21）： 86-90.
[14]	贺文华，董晓慧，汤臣建，等. “宗筋主束骨而利机关”理论在经筋病中的临床应用概况［J］. 湖南中医杂志， 2019， 35（5）： 155-157.
[15]	COLETTI C， ACOSTA G F， KESLACY S， et al. Exercise-mediated reinnervation of skeletal muscle in elderly people： an update［J］. Eur J Transl Myol， 2022， 32（1）： 10416.
[16]	ZHENG D D， LIU J， PIAO H L， et al. ROS-triggered endothelial cell death mechanisms： Focus on pyroptosis， parthanatos， and ferroptosis［J］. Front Immunol， 2022， 13： 1039241.
[17]	黄吴心蕊，田静，贺洁宇. 原发性干燥综合征合并骨质疏松的危险因素［J］.中南大学学报（医学版）， 2024， 49（2）： 312-318.
[18]	SOUSA-VICTOR P， GARCÍA-PRAT L， MUÑOZ-CÁNOVES P. Control of satellite cell function in muscle regeneration and its disruption in ageing［J］. Nat Rev Mol Cell Biol， 2022， 23（3）： 204-226.
[19]	KUANG S H， KURODA K， LE GRAND F， et al. Asymmetric self-renewal and commitment of satellite stem cells in muscle［J］. Cell， 2007， 129（5）： 999-1010.
[20]	BACHMAN J F， CHAKKALAKAL J V. Satellite cells in the growth and maintenance of muscle［J］. Curr Top Dev Biol， 2024， 158： 1-14.
[21]	COLLINS B C， KARDON G. It takes all kinds： heterogeneity among satellite cells and fibro-adipogenic progenitors during skeletal muscle regeneration［J］. Development， 2021， 148（21）： dev199861.
[22]	PIETROSEMOLI N， MELLA S， YENNEK S， et al. Comparison of multiple transcriptomes exposes unified and divergent features of quiescent and activated skeletal muscle stem cells［J］. Skeletal Muscle， 2017， 7（1）： 28.
[23]	SU Y， YU Y Y， LIU C C， et al. Fate decision of satellite cell differentiation and self-renewal by miR-31-IL34 axis［J］. Cell Death Differ， 2020， 27（3）： 949-965.
[24]	SONG Z W， JIN C L， YE M， et al. Lysine inhibits apoptosis in satellite cells to govern skeletal muscle growth via the JAK2-STAT3 pathway［J］. Food Funct， 2020， 11（5）： 3941-3951.

Group	Circumference（l/mm）	Wet weight （m/g）	Wet weight maintenance rate（η/%）	Ratio of wet weight/body mass
Control	157.44±11.88	5.17±0.84	95.78±2.17	4.78±0.67
Model	99.33±11.16^*	3.00±0.50^*	82.67±5.20^*	2.33±0.50^*
Celecoxib	105.56±13.51^△	3.78±0.67^△	86.67±3.67^△	3.00±0.87^△
Acupuncture	134.22±16.52^△#	4.56±0.53^△#	94.33±2.69^△#	4.56±0.73^△#

Group	Apoptosis index
Group	Articular cartilage	Quadriceps femoris muscle tissue
Control	0.62±0.10	0.78±0.19
Model	1.52±0.29^*	2.42±0.37^*
Celecoxib	1.34±0.19^△	1.56±0.24^△
Acupuncture	1.03±0.21^△#	0.91±0.18^△#

Group	IL-6	JAK	STAT3
Control	12.04±2.12	10.02±1.73	9.83±1.45
Model	1.02±0.21^*	1.45±0.31^*	2.72±0.36^*
Celecoxib	5.13±1.25^△	6.23±1.12^△	5.24±1.19^△
Acupuncture	9.14±1.32^△#	8.07±1.53^△#	9.02±0.54^△#

Group	IL-6	JAK	STAT3
Control	0.73±0.13	0.83±0.13	0.98±0.16
Model	0.12±0.08^*	0.44±0.10^*	0.21±0.09^*
Celecoxib	0.52±0.10^△	0.53±0.12^△	0.76±0.10^△
Acupuncture	0.80±0.15^△#	0.76±0.17^△#	0.81±0.12^△#

Group	Pax7	Desmin	Myosin	Myogenin
Control	0.87±0.13	0.72±0.13	0.90±0.07	0.81±0.12
Model	0.15±0.03^*	0.10±0.05^*	0.14±0.04^*	0.24±0.08^*
Celecoxib	0.67±0.10^△	0.51±0.12^△	0.49±0.15^△	0.62±0.17^△
Acupuncture	0.72±0.16^△#	0.70±0.13^△#	0.74±0.12^△#	0.83±0.21^△#

Effect of acupuncture on differentiation and apoptosis of quadriceps muscle satellite cells in knee osteoarthritis model rats and its mechanism

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Group	Bcl-2	Bcl-xl	Bax	MCL1	Caspase-3
Control	0.91±0.13	0.94±0.14	0.13±0.02	0.70±0.13	0.09±0.01
Model	0.13±0.02^*	0.10±0.02^*	0.92±0.10^*	0.15±0.02^*	0.93±0.12^*
Celecoxib	0.21±0.04^△	0.31±0.07^△	0.48±0.12^△	0.23±0.03^△	0.62±0.13^△
Acupuncture	0.83±0.12^△#	0.78±0.12^△#	0.19±0.04^△#	0.73±0.12^△#	0.36±0.06^△#

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