Gang Zhong1, Huiping Long2, Shiting Ma3, Yao Shunhan4, Jia Li5, Jun Yao6. 1. Department of Bone and Joint Surgery, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China; Guangxi Collaborative Innovation Center for Biomedicine, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China. Electronic address: 13257788667@163.com. 2. Guangxi Medical University, 530021, Nanning, China. 3. Department of Bone and Joint Surgery, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China; Guangxi Collaborative Innovation Center for Biomedicine, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China. 4. Department of Bone and Joint Surgery, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China. 5. Department of Pathology, First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China. Electronic address: jialee2005@126.com. 6. Department of Bone and Joint Surgery, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China; Guangxi Collaborative Innovation Center for Biomedicine, The First Affiliated Hospital of Guangxi Medical University, 530021 Nanning, China. Electronic address: gxyaojun@163.com.
Abstract
AIMS: Osteoarthritis (OA) is a chronic and degenerative joint disease prevalent in the elderly, which is characterized by hypertrophy and reactive hyperplasia of articular cartilage. Autophagy has been reported to inhibit inflammation and reduce chondrocyte apoptosis in OA. As the microRNA (miRNA)-335-5p has been linked to both inflammation and autophagy, this study aimed to investigate its potential role in regulating autophagy during the pathogenesis of OA. MAIN METHODS: Quantitative real-time PCR (qRT-PCR) was used to detect miRNA-335-5p expression in normal and OA human chondrocytes. Following transfection of human OA chondrocytes with double-stranded miRNA-335-5p mimic/inhibitor, qRT-PCR, western blotting, and immunofluorescence were used to determine expression levels of the inflammatory mediators IL-1β, IL-6, and TNF-α, and the autophagic markers Beclin-1, autophagy-related protein 5 (ATG5), and ATG7. The autophagy inhibitor 3-methyladenine (3-MA) was used to link the anti-inflammatory effects of miRNA-335-5p to autophagy. KEY FINDINGS: The expression of miRNA-335-5p was significantly lower in OA chondrocytes than in normal chondrocytes. Transfection of human OA chondrocytes with the miRNA-335-5p mimic led to a remarkable increase in viability, a significant increase in autophagy-related factors, and a reduction in inflammatory mediators. Importantly, treatment of miRNA-335-5p-overexpressing OA chondrocytes with the autophagy inhibitor 3-MA restored the expression of inflammatory mediators. SIGNIFICANCE: We conclude that miRNA-335-5p can significantly alleviate inflammation in human OA chondrocytes by activating autophagy. Therefore, miRNA-335-5p has potential for future use in the clinical diagnosis and treatment of OA.
AIMS: Osteoarthritis (OA) is a chronic and degenerative joint disease prevalent in the elderly, which is characterized by hypertrophy and reactive hyperplasia of articular cartilage. Autophagy has been reported to inhibit inflammation and reduce chondrocyte apoptosis in OA. As the microRNA (miRNA)-335-5p has been linked to both inflammation and autophagy, this study aimed to investigate its potential role in regulating autophagy during the pathogenesis of OA. MAIN METHODS: Quantitative real-time PCR (qRT-PCR) was used to detect miRNA-335-5p expression in normal and OA human chondrocytes. Following transfection of human OA chondrocytes with double-stranded miRNA-335-5p mimic/inhibitor, qRT-PCR, western blotting, and immunofluorescence were used to determine expression levels of the inflammatory mediators IL-1β, IL-6, and TNF-α, and the autophagic markers Beclin-1, autophagy-related protein 5 (ATG5), and ATG7. The autophagy inhibitor 3-methyladenine (3-MA) was used to link the anti-inflammatory effects of miRNA-335-5p to autophagy. KEY FINDINGS: The expression of miRNA-335-5p was significantly lower in OA chondrocytes than in normal chondrocytes. Transfection of human OA chondrocytes with the miRNA-335-5p mimic led to a remarkable increase in viability, a significant increase in autophagy-related factors, and a reduction in inflammatory mediators. Importantly, treatment of miRNA-335-5p-overexpressing OA chondrocytes with the autophagy inhibitor 3-MA restored the expression of inflammatory mediators. SIGNIFICANCE: We conclude that miRNA-335-5p can significantly alleviate inflammation in human OA chondrocytes by activating autophagy. Therefore, miRNA-335-5p has potential for future use in the clinical diagnosis and treatment of OA.