通过调节软骨细胞有丝分裂 Pink1-Parkin 通路缓解兔膝关节骨关节炎

Zhu Wenting, Guo Changqing, D U Mei, M A Yunxuan, Cui Yongqi, Chen Xilin, Guo Changqing
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摘要

目的研究穴位切除术对膝关节骨关节炎(KOA)兔软骨细胞有丝分裂和 Pink1-Parkin 通路的影响:方法:通过改良 Videman 法建立 KOA 模型。兔子被随机分为对照组(CON)、KOA 组和 KOA + 穴位切除组(Acu)。穴位切除组的兔子在模型建立后接受穴位切除术 4 周。记录干预前后兔子的行为。用光学显微镜和荧光显微镜评估软骨退化情况。透射电子显微镜、免疫荧光和酶联免疫吸附试验(ELISA)评估了有丝分裂的水平。免疫荧光、Western 印迹和实时聚合酶链反应评估了磷酸酶和天丝同源物(PTEN)诱导的激酶 1(Pink1)-Parkin 有丝分裂途径成分的表达:结果:KOA兔出现关节疼痛、活动障碍和软骨退行性变,Mankin评分升高,Ⅱ型胶原(Col-Ⅱ)表达显著下降,有丝分裂受抑制,线粒体功能受损,Pink1-Parkin通路相关因子受抑制。穴位切除术调节了Pink1-Parkin通路相关蛋白、有丝分裂相关蛋白微管相关蛋白-1轻链-3、外膜易位酶和线粒体内膜23的表达;增加了线粒体和自噬体的共定位;促进了受损线粒体的清除;恢复了线粒体三磷酸腺苷(ATP)的产生;缓解了KOA兔的软骨退化:通过调节与 Pink1-Parkin 通路相关的蛋白,激活软骨细胞中的有丝分裂,针刺术在缓解家兔 KOA 中发挥了作用。
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Acupotomy alleviates knee osteoarthritis in rabbit by regulating chondrocyte mitophagy Pink1-Parkin pathway.

Objective: To investigate the effect of acupotomy, on mitophagy and the Pink1-Parkin pathway in chondrocytes from rabbits with knee osteoarthritis (KOA).

Methods: A KOA model was established via the modified Videman method. Rabbits were randomly divided into a control group (CON), KOA group and KOA + acupotomy group (Acu). Rabbits in the acupotomy group were subjected to acupotomy for 4 weeks after model establishment. The behavior of the rabbits before and after intervention was recorded. Cartilage degeneration was evaluated by optical microscopy and fluorescence microscopy. The level of mitophagy was evaluated by transmission electron microscopy, immunofluorescence and enzyme-linked immunosorbent assay (ELISA). The expression of phosphatase and tensin homolog (PTEN)-induced kinase 1 (Pink1)-Parkin mitophagy pathway components was evaluated by immunofluorescence, Western blotting and real-time polymerase chain reaction.

Results: In rabbits with KOA, joint pain, mobility disorders and cartilage degeneration were observed, the Mankin score was increased, collagen type Ⅱ (Col-Ⅱ) expression was significantly decreased, mitophagy was inhibited, mitochondrial function was impaired, and factors associated with the Pink1-Parkin pathway were inhibited. Acupotomy regulated the expression of Pink1-Parkin pathway-related proteins, the mitophagy-related protein microtubule-associated protein-1 light chain-3, the translocase of the outer membrane, and the inner mitochondrial membrane 23; increased the colocalization of mitochondria and autophagosomes; promoted the removal of damaged mitochondria; restored mitochondrial adenosine-triphosphate (ATP) production; and alleviated cartilage degeneration in rabbits with KOA.

Conclusions: Acupotomy played a role in alleviating KOA in rabbits by activating mitophagy in chondrocytes via the regulation of proteins that are related to the Pink1-Parkin pathway.

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