中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (14): 2882-2891.doi: 10.12307/2025.387

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

蠲痹汤含药血清调控线粒体自噬抑制白细胞介素1β诱导的关节软骨细胞损伤

郑永智1，陈飞飞2，康  乾1，晋春阳1，王若秦1   

  1. 1河南省中医院关节病科，河南省郑州市  450000；2河南省胸科医院结核内科，河南省郑州市  450000


  • 收稿日期:2024-02-27 接受日期:2024-06-01 出版日期:2025-05-18 发布日期:2024-09-27
  • 通讯作者: 郑永智，硕士，副主任医师，河南省中医院关节病科，河南省郑州市 450000
  • 作者简介:郑永智，男，1978年生，河南省长垣县人，汉族，2007年昆明医学院毕业，硕士，副主任医师，主要从事骨关节疾病的中西医防治进展的研究。
  • 基金资助:
    河南省教育厅厅局级项目(23A360023)，项目负责人：郑永智

Juanbi Decoction-containing serum inhibits interleukin-1beta induced articular chondrocyte damage via regulating mitophagy

Zheng Yongzhi1, Chen Feifei2, Kang Qian1, Jin Chunyang1, Wang Ruoqin1   

  1. 1Department of Arthropathy, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, Henan Province, China; 2Department of Tuberculosis, Henan Provincial Chest Hospital, Zhengzhou 450000, Henan Province, China
  • Received:2024-02-27 Accepted:2024-06-01 Online:2025-05-18 Published:2024-09-27
  • Contact: Zheng Yongzhi, Master, Associate chief physician, Department of Arthropathy, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, Henan Province, China
  • About author:Zheng Yongzhi, Department of Arthropathy, Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, Henan Province, China
  • Supported by:
    Henan Provincial Department of Education Bureau-level Project, No. 23A360023 (to ZYZ)

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摘要：



文题释义：
蠲痹汤：由羌活、独活、肉桂、秦艽、海风藤、桑枝、当归、川芎、乳香、木香、甘草等药物组成，具有祛风除湿、蠲痹止痛之功效，对于缓解全身关节疼痛和改善活动功能障碍有确切疗效。
线粒体自噬：细胞会通过自噬机制来选择性地清除受损线粒体，来维持线粒体的稳态和线粒体质量。软骨细胞线粒体自噬的缺陷会引起细胞凋亡和基质消失等软骨细胞退行性病变的变化。因此，维持细胞线粒体的稳态是治疗膝骨关节炎的潜在有效策略。

背景：软骨细胞线粒体自噬的缺陷会引起细胞凋亡和基质消失等软骨细胞退行性病变的变化。
目的：探讨蠲痹汤含药血清对白细胞介素1β诱导的大鼠膝关节软骨细胞炎症反应和凋亡的影响及可能作用机制。
方法：50只雄性SD大鼠随机给予生理盐水、蠲痹汤低、中、高剂量(1.24，2.48，4.96 g/kg)、塞来昔布(阳性药物)，连续灌胃2周后获得含药血清。①分离软骨细胞，将其随机分为对照组、白细胞介素1β组、蠲痹汤低、中、高剂量含药血清组及阳性药物血清组。CCK-8法检测细胞存活率、免疫荧光双染检测线粒体自噬水平、免疫荧光检测磷酸化腺苷酸激活蛋白激酶水平、Western blot检测PTEN诱导激酶1/Parkin通路相关蛋白和裂解的半胱氨酸蛋白酶蛋白3表达、ELISA检测炎症因子水平；②分别采用PTEN诱导激酶1 siRNA和Compound C进行干预，探究AMPK/PTEN诱导激酶1/Parkin通路在蠲痹汤含药血清调控线粒体自噬中的作用。
结果与结论：①与对照组比较，白细胞介素1β组软骨细胞存活率、Ⅱ型胶原蛋白表达、磷酸化腺苷酸激活蛋白激酶、PTEN诱导激酶1、Parkin和微管相关蛋白1轻链3蛋白水平以及线粒体自噬水平明显降低(P < 0.05)，而裂解的半胱氨酸蛋白酶蛋白3蛋白水平、白细胞介素6、白细胞介素8和肿瘤坏死因子α水平显著升高(P < 0.05)；与白细胞介素1β组比较，蠲痹汤各剂量含药血清组和阳性药物血清组上述各项指标呈现相反的变化(P < 0.05)；②PTEN诱导激酶1 siRNA可显著抑制蠲痹汤含药血清对白细胞介素1β处理软骨细胞线粒体自噬的影响，降低蠲痹汤含药血清对白细胞介素1β诱导的软骨细胞炎症与凋亡的保护作用；Compound C逆转了蠲痹汤含药血清对白细胞介素1β处理软骨细胞中PTEN诱导激酶1/Parkin信号通路的影响。结论：蠲痹汤含药血清通过影响线粒体自噬水平来抑制软骨细胞炎症和凋亡，从而减轻白细胞介素1β诱导的软骨细胞退化，其机制可能与调控AMPK/PTEN诱导激酶1/Parkin通路有关。
https://orcid.org/0009-0002-1188-8300(郑永智)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 蠲痹汤, 软骨细胞, 线粒体自噬, 腺苷酸激活蛋白激酶, AMPK, PTEN诱导激酶1/Parkin

Abstract: BACKGROUND: Defective outgrowth of chondrocyte mitophagy causes degenerative chondrocyte pathological changes such as apoptosis and matrix loss. 
OBJECTIVE: To investigate the effects and possible mechanism of Juanbi Decoction-containing serum on interleukin-1β-induced inflammatory response and apoptosis of rat knee joint chondrocytes. 
METHODS: Fifty male Sprague-Dawley rats were randomly given saline, Juanbi Decoction in low, medium and high doses (1.24, 2.48 and 4.96 g/kg), and celecoxib (positive drug) group. Drug-containing serum was obtained by continuous gavage for 2 weeks. (1) Chondrocytes were isolated and randomly divided into control group, interleukin-1β group, Juanbi Decoction low, medium and high dose containing serum groups and positive drug serum group. Cell counting kit-8 assay was used to detect cell viability; immunofluorescence double staining was used to detect mitophagy level; immunofluorescence was used to detect the phosphorylated AMP-activated protein kinase level; western blot was used to detect the expression of PTEN induced putative kinase 1/Parkin pathway-related protein and cleaved caspase-3; and ELISA was used to detect inflammatory factor level. (2) PTEN induced putative kinase 1 siRNA and Compound C were used for intervention to explore the role of AMP-activated protein kinase/PTEN induced putative kinase 1/Parkin pathway in the regulation of mitophagy by Juanbi Decoction-containing serum. 
RESULTS AND CONCLUSION: (1) Compared with the control group, chondrocyte survival, type II collagen, phosphorylated AMP-activated protein kinase, PTEN induced putative kinase 1, Parkin, and microtubule-associated proteins 1 light chain 3 protein levels, and mitophagy levels were significantly lower in the interleukin-1β group (P < 0.05), while cleaved caspase-3 protein levels, interleukin-6, interleukin-8 and tumor necrosis factor-α levels were significantly higher (P < 0.05). Compared with the interleukin-1β group, opposite changes in all the above indexes were observed in Juanbi Decoction low, medium and high dose containing serum groups and positive drug serum group (P < 0.05). (2) PTEN induced putative kinase 1 siRNA significantly inhibited the effect of Juanbi Decoction-containing serum on mitophagy in interleukin-1β-treated chondrocytes and reduced the protective effect of Juanbi Decoction-containing serum on interleukin-1β-induced inflammation and apoptosis in chondrocytes; Compound C reversed the effect of Juanbi Decoction-containing serum on the PTEN induced putative kinase 1/Parkin signaling pathway in interleukin-1β-treated chondrocytes. To conclude, Juanbi Decoction-containing serum inhibits chondrocyte inflammation and apoptosis by affecting the level of mitochondrial autophagy, thereby attenuating interleukin-1β-induced chondrocyte degradation by a mechanism that may be related to the regulation of the AMP-activated protein kinase/PTEN induced putative kinase 1/Parkin pathway.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Juanbi Decoction, chondrocyte, mitophagy, adenylate-activated protein kinase, AMPK, PTEN induced putative kinase 1/Parkin

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