Min Qiu1, Da Liu2, Qin Fu3. 1. Department of Orthopedics, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning, China. Electronic address: qiumin834@163.com. 2. Department of Gynaecology and Obstetrics, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning, China. 3. Department of Orthopedics, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning, China.
Abstract
AIMS: To explore the therapeutic effect of miR-129-5p carried by exosomes from Human Synovial Mesenchymal Stem Cell (HS-MSC) on osteoarthritis(OA). MATERIALS AND METHODS: The levels of miR-129-5p and high mobility group protein -1 (HMGB1) and interleukin-1β (IL-1β) in the joint fluid of OA patients were respectively detected via real-time quantitative reverse transcription-PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-1β was taken to act on chondrocytes for the establishment of OA model in vitro. Ultracentrifugation was conducted to isolate HS-MSC exosomes (HS-MSC-Exo) from the supernatant. Western blot and ELISA were carried out to measure the expression of iNOS, COX2, MMP13, Collagen 2, TLR4, NF-κB, Caspase3, Bcl-2, HMGB1 in chondrocytes. Flow cytometry was conducted to detect the apoptosis of chondrocytes. Besides, bioinformatics was employed to predict the targeted relationship between miR-129-5p and HMGB1, which was further verified via dual luciferase activity experiments. KEY FINDINGS: The results illustrated that miR-129-5p was decreased in OA patients and IL-1β-induced chondrocytes, while HMGB1 was notably upregulated. HS-MSC-Exo rich in miR-129-5p remarkably declined the inflammatory response and apoptosis of chondrocytes, while HS-MSC-Exo deficient in miR-129-5p increased the IL-1β-mediated inflammatory response and apoptosis of chondrocytes. In terms of mechanism, miR-129-5p targets the 3'UTR end of HMGB1 and inhibits IL-1β-mediated upregulation of HMGB1. SIGNIFICANCE: In a word, this paper proved that miR-129-5p, existing in HS-MSC-Exo, can suppress the IL-1β-mediated OA by inhibiting HMGB1 release.
AIMS: To explore the therapeutic effect of miR-129-5p carried by exosomes from Human Synovial Mesenchymal Stem Cell (HS-MSC) on osteoarthritis(OA). MATERIALS AND METHODS: The levels of miR-129-5p and high mobility group protein -1 (HMGB1) and interleukin-1β (IL-1β) in the joint fluid of OA patients were respectively detected via real-time quantitative reverse transcription-PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-1β was taken to act on chondrocytes for the establishment of OA model in vitro. Ultracentrifugation was conducted to isolate HS-MSC exosomes (HS-MSC-Exo) from the supernatant. Western blot and ELISA were carried out to measure the expression of iNOS, COX2, MMP13, Collagen 2, TLR4, NF-κB, Caspase3, Bcl-2, HMGB1 in chondrocytes. Flow cytometry was conducted to detect the apoptosis of chondrocytes. Besides, bioinformatics was employed to predict the targeted relationship between miR-129-5p and HMGB1, which was further verified via dual luciferase activity experiments. KEY FINDINGS: The results illustrated that miR-129-5p was decreased in OA patients and IL-1β-induced chondrocytes, while HMGB1 was notably upregulated. HS-MSC-Exo rich in miR-129-5p remarkably declined the inflammatory response and apoptosis of chondrocytes, while HS-MSC-Exo deficient in miR-129-5p increased the IL-1β-mediated inflammatory response and apoptosis of chondrocytes. In terms of mechanism, miR-129-5p targets the 3'UTR end of HMGB1 and inhibits IL-1β-mediated upregulation of HMGB1. SIGNIFICANCE: In a word, this paper proved that miR-129-5p, existing in HS-MSC-Exo, can suppress the IL-1β-mediated OA by inhibiting HMGB1 release.
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