Lei Fan1, Ming Li2, Fu-Yang Cao2, Zhi-Wei Zeng2, Xiao-Bin Li1, Chao Ma3, Jing-Tao Ru4, Xue-Jian Wu5. 1. Department of Orthopedic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450002, China; Department of Orthopedic Surgery, People's Hospital of Zhengzhou University, Zhengzhou 450003, China. 2. Basic Medical College of Zhengzhou University, Zhengzhou 450001, China. 3. Department of Cardiology, Campus Virchow, Charité-Universitätsmedizin Berlin, Berlin 13353, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Berlin, Berlin, Germany. Electronic address: chao.ma@charite.de. 4. Department of Orthopedic Surgery, People's Hospital of Zhengzhou University, Zhengzhou 450003, China. Electronic address: rengcaitan5576kt@163.com. 5. Department of Orthopedic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450002, China. Electronic address: wu14xuejian@sina.com.
Abstract
BACKGROUND: Astragalus polysaccharide (APS) is a traditional Chinese herbal medicine with anti-inflammatory and anti-aging activities. OBJECTIVE: This study aimed to explore the effect and associated mechanisms of APS on LPS-induced injury in ATDC5 cells, to evaluate the potential of APS for use as an adjuvant therapy for osteoarthritis (OA). MATERIALS AND METHODS: ATDC5 cells were pre-treated with APS and stimulated with lipopolysaccharide (LPS). Cell viability, ROS generation as well as the expression of IL-6, TNF-α, iNOS and Cox-2 were evaluated by performing CCK8 assay, ROS detection, ELISA, western blot and qRT-PCR. The expression of NF-κB and p38MAPK signal pathways related proteins and KLF4 was measured through western blot. RESULTS: LPS increased the expression of IL-6 and TNF-α, elevated the expression of Cox-2, iNOS and increased ROS generation. APS treatment significantly alleviated LPS-induced damage in ATDC5 cells. Besides, miR-92a was down-regulated while KLF4 was up-regulated by APS. At the same time, the targeting relationship between miR-92a and KLF4 was demonstrated. The inhibitory effects of APS on LPS-induced injury in ATDC5 cells were attenuated by the combination of KLF4 siRNA. In addition, LPS induced NF-κB and p38MAPK signal pathways were decreased by APS treatment. Also, the inhibitory effect of APS on NF-κB and p38MAPK signal pathways was reversed by KLF4 siRNA. CONCLUSIONS: The present study reveals that APS protects ATDC5 cells against LPS induced-injury by regulation of miR-92a/KLF4 axis and suppressing NF-κB and p38MAPK signal pathways.
BACKGROUND: Astragalus polysaccharide (APS) is a traditional Chinese herbal medicine with anti-inflammatory and anti-aging activities. OBJECTIVE: This study aimed to explore the effect and associated mechanisms of APS on LPS-induced injury in ATDC5 cells, to evaluate the potential of APS for use as an adjuvant therapy for osteoarthritis (OA). MATERIALS AND METHODS: ATDC5 cells were pre-treated with APS and stimulated with lipopolysaccharide (LPS). Cell viability, ROS generation as well as the expression of IL-6, TNF-α, iNOS and Cox-2 were evaluated by performing CCK8 assay, ROS detection, ELISA, western blot and qRT-PCR. The expression of NF-κB and p38MAPK signal pathways related proteins and KLF4 was measured through western blot. RESULTS: LPS increased the expression of IL-6 and TNF-α, elevated the expression of Cox-2, iNOS and increased ROS generation. APS treatment significantly alleviated LPS-induced damage in ATDC5 cells. Besides, miR-92a was down-regulated while KLF4 was up-regulated by APS. At the same time, the targeting relationship between miR-92a and KLF4 was demonstrated. The inhibitory effects of APS on LPS-induced injury in ATDC5 cells were attenuated by the combination of KLF4 siRNA. In addition, LPS induced NF-κB and p38MAPK signal pathways were decreased by APS treatment. Also, the inhibitory effect of APS on NF-κB and p38MAPK signal pathways was reversed by KLF4 siRNA. CONCLUSIONS: The present study reveals that APS protects ATDC5 cells against LPS induced-injury by regulation of miR-92a/KLF4 axis and suppressing NF-κB and p38MAPK signal pathways.