Effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis based on PI3K/Akt/mTOR signaling pathway

LI Dekun△; YAO Changfeng; SHAN Ziliang; ZHOU Zheng; ZHANG Xianji; WANG Kewen; DU Shaolin

doi:10.13703/j.0255-2930.20241026-k0002

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Effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis based on PI3K/Akt/mTOR signaling pathway

Study on Mechanism | 更新时间：2025-10-11

- Effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis based on PI3K/Akt/mTOR signaling pathway
- Chinese Acupuncture & Moxibustion Vol. 45, Issue 10, Pages: 1459-1467(2025)
- 作者机构：
  
  安徽中医药大学针灸推拿学院，合肥　230012
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20241026-k0002
  CLC：
- Received：26 October 2024，
  
  Online First：14 March 2025，
  
  Published：12 October 2025
- 稿件说明：
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LI Dekun△, YAO Changfeng, SHAN Ziliang, et al. Effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis based on PI3K/Akt/mTOR signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(10): 1459-1467.
DOI：

LI Dekun△, YAO Changfeng, SHAN Ziliang, et al. Effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis based on PI3K/Akt/mTOR signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(10): 1459-1467. DOI： 10.13703/j.0255-2930.20241026-k0002.

摘要

目的：

观察针刺对膝关节骨关节炎（KOA）大鼠膝关节软骨自噬的影响，并探讨其作用机制。

方法：

将40只SPF级雄性SD大鼠随机分为空白组、模型组、悬吊组、针刺组、联合组，每组8只。除空白组外，其余组大鼠采用木瓜蛋白酶注射法建立KOA模型。悬吊组、针刺组、联合组分别行悬吊运动疗法（每次10 min，每日3次）、针刺（穴取右侧“阳陵泉”“足三里”“犊鼻”，每次30 min，每日1次）及联合疗法（悬吊运动疗法联合针刺）进行干预，连续6 d后休息1 d，持续4周。干预前后评估大鼠Lequesne MG评分。干预后，采用HE染色法观察大鼠右膝关节软骨组织形态，并评定Mankin评分；ELISA法检测大鼠血清白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α（TNF-α）含量；实时荧光定量PCR法检测大鼠右膝关节软骨组织Ⅱ型胶原蛋白（COL2）、Ⅹ型胶原蛋白（COL10）、磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B（Akt）、哺乳动物雷帕霉素靶蛋白（mTOR）、自噬调控蛋白（Beclin-1）mRNA表达；Western blot法检测大鼠右膝关节软骨组织PI3K、磷酸化PI3K（p-PI3K）、Akt、磷酸化Akt（p-Akt）、mTOR、磷酸化mTOR（p-mTOR）、Beclin-1蛋白表达。

结果：

与空白组比较，模型组大鼠Lequesne MG评分升高（

0.01）；右膝关节软骨厚度变薄，软骨细胞数量减少且排列杂乱，Mankin评分升高（

0.01）；血清IL-1β、IL-6、TNF-α含量升高（

0.01）；右膝关节软骨组织COL2、Beclin-1 mRNA及Beclin-1蛋白表达降低（

0.01），COL10、PI3K、Akt、mTOR mRNA及p-PI3K、p-Akt、p-mTOR蛋白表达升高（

0.01）。与模型组比较，悬吊组、针刺组和联合组大鼠Lequesne MG评分降低（

0.01）；右膝关节软骨厚度增加，软骨细胞排列改善，Mankin评分降低（

0.01）；血清IL-1β、IL-6、TNF-α含量降低（

0.01）；右膝关节软骨组织COL2、Beclin-1 mRNA及Beclin-1蛋白表达增加（

0.05，

0.01），COL10、PI3K、Akt、mTOR mRNA及p-PI3K、p-Akt、p-mTOR蛋白表达降低（

0.05，

0.01）。与悬吊组、针刺组比较，联合组大鼠Lequesne MG评分降低（

0.01）；Mankin评分降低（

0.05，

0.01）；血清IL-1β、IL-6、TNF-α含量降低（

0.05）；右膝关节软骨组织COL2、Beclin-1 mRNA及Beclin-1蛋白表达增加（

0.05），COL10、PI3K、Akt、mTOR mRNA及p-PI3K、p-Akt、p-mTOR蛋白表达降低（

0.05，

0.01）。

结论：

针刺可以促进KOA大鼠膝关节软骨修复和细胞自噬，降低炎性反应，从而缓解软骨退变，作用机制可能与调节PI3K/Akt/mTOR信号通路有关。

Abstract

Objective

To observe the effect of acupuncture on chondrocyte autophagy in rats of knee osteoarthritis (KOA) and explore its underlying mechanisms.

Methods

Forty male SPF-grade SD rats were randomized into a blank group

a model group

a suspension group

an acupuncture group

and a combined therapy group

8 rats in each one. Except the blank group

KOA model was prepared by the injection with papain. The suspension exercise therapy (10 min each time

three times daily)

acupuncture (at "Yanglingquan" [GB34]

"Zusanli" [ST36]

and "Dubi" [ST35] on the right side

30 min each intervention

once daily) and the combined therapy (the suspension exercise therapy combined with acupuncture) were delivered in the suspension group

the acupuncture group and the combined therapy group

respectively. The intervention of each group was performed continuously for 6 days

and 4 consecutive weeks

at the interval of 1 day. Before and after intervention

Lequesne MG score was assessed in the rats. After intervention

HE staining was adopted to observe the cartilaginous tissue morphology of the right knee joints

and Mankin score was evaluated; the serum levels of interleukin (IL)-1β

IL-6

and tumor neurosis factor-α (TNF-α) were measured using ELISA; the real-time PCR was provided to determine the mRNA expression of collagen protein type Ⅱ(COL2)

collagen protein type Ⅹ (COL10)

phosphatidylinositol 3-kinase (PI3K)

protein kinase B (Akt)

mammalian target of rapamycin (mTOR)

and autophagy-regulated protein (Beclin-1) in the cartilaginous tissue of the right knee joint; Western blot was employed to detect the protein expression of PI3K

phosphorylated PI3K (p-PI3K)

Akt

phosphorylated Akt (p-Akt)

mTOR

phosphorylated mTOR (p-mTOR) and Beclin-1 in the cartilaginous tissue of the right knee joint.

Results

Compared with the blank group

the rats in the model group showed the higher Lequesne MG score (

0.01)

thinner cartilage of the right knee

reduced chondrocytes and disordered arrangement

and higher Mankin score (

0.01). Besides

in the model group

the serum levels of IL-1β

IL-6 and TNF-α were elevated (

0.01)

the mRNA expression of COL2 and Beclin-1 and the protein expression of Beclin-1 decreased (

0.01)

the mRNA expression of COL10

PI3K

Akt and mTOR

and the protein expression of p-PI3K

p-Akt and p-mTOR increased (

0.01) in the cartilaginous tissue of the right knee joint. Compared with the model group

in the suspension group

the acupuncture group and the combined therapy group

the Lequesne MG scores were reduced (

0.01)

the cartilage of the right knee was thickened

the arrangement of chondrocytes was improved

and the Mankin scores were lower (

0.01). Besides

in these intervention groups

the serum levels of IL-1β

IL-6 and TNF-α were reduced (

0.01)

the mRNA expression of COL2 and Beclin-1 and the protein expression of Beclin-1 increased (

0.05

0.01)

the mRNA expression of COL10

PI3K

Akt and mTOR

and the protein expre

ssion of p-PI3K

p-Akt and p-mTOR decreased (

0.05

0.01) in the cartilaginous tissue of the right knee joint. When compared with the suspension group and the acupuncture group

in the combined therapy group

the Lequesne MG score was reduced (

0.01)

and the Mankin score was reduced (

0.05

0.01). Besides

the serum levels of IL-1β

IL-6 and TNF-α were reduced (

0.05)

the mRNA expression of COL2 and Beclin-1 and the protein expression of Beclin-1 increased (

0.05)

the mRNA expression of COL10

PI3K

Akt and mTOR

and the protein expression of p-PI3K

p-Akt and p-mTOR decreased (

0.05

0.01) in the cartilaginous tissue of the right knee joint.

Conclusion

Acupuncture can promote cartilage regeneration of knee joint and autophagy in KOA rats

alleviate inflammation

so as to retard cartilage degeneration

which may be possibly associated with the PI3K/Akt/mTOR signaling pathway.

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