miR-98-5p inhibits the development of osteoarthritis through STAT3 pathway
Objective To explore the possible molecular mechanism of miR-98-5p regulating the occurrence and development of osteo-arthritis(OA).Methods ①lRat chondrocytes were divided into control group,interleukin-1 β(IL-1 β)group,NC-agomir group,miR-98-5p-agomir group,NC-antagomir group and miR-98-5p-antagomir group.The chondrocytes were cultured with normal DMEM medium in control group,and the cells in other groups were cultured in the medium supplemented with 10 ng/mL IL-1 β for 24 h after successful transfection.The proliferation and the apoptosis of chondrocytes were detected by MTT and TUNEL methods,respectively.The transcriptional levels of miR-98-5p,STAT3,Bcl-2,Bax,IL-1β,IL-6 and TNF-α were detected by RT-qPCR.The levels of IL-1 β,IL-6 and TNF-α in cell culture supernatant were detected by ELISA.The levels of Bcl-2,Bax,p-STAT3 and STAT3 proteins were detected by Western blot.②OA rat model was established by transection of anterior cruciate ligament and partial resection of medial meniscus.After modeling,the OA rats were divided into OA group,NC-agomir+OA group and miR-98-5p-agomir+OA group,with 12 rats in each group.Another 12 healthy rats were chosen as sham group.Totally 40 μL normal saline was injected into the articular cavity of rats in sham group and OA group,while 40 μL NC-agomir and miR-98-5p-agomir(1 nmol/μL)were injected into the articular cavity in NC-agomir+OA group and miR-98-5p-agomir+OA group,respectively.The injection was given once a week for four weeks.The cartilage degeneration of rats was observed by Safranin O-fast green staining.Apoptosis of chondrocytes was observed by TUNEL staining.The transcriptional levels of miR-98-5p,STAT3,Bcl-2,Bax,IL-1β,IL-6 and TNF-α in cartilage were detected by RT-qPCR.The levels of IL-1β,IL-6 and TNF-α in cartilage tissue were detected by ELISA.The levels of Bcl-2,Bax,p-STAT3 and STAT3 protein in cartilage were detected by Western blot.Results ①Compared with IL-1β group and NC-agomir group,the relative cell viability,Bcl-2 mRNA and protein levels and miR-98-5p level were increased in miR-98-5p-agomir group(P＜0.05),while TUNEL positive rate,the mRNA and protein levels of Bax,IL-1 β,IL-6,TNF-α and STAT3,and the protein level of p-STAT3 and p-STAT3/STAT3 ratio were decreased(P＜0.05).Compared with IL-1β group and NC-antagomir group,the relative cell viability,Bcl-2 mRNA and protein levels and miR-98-5p level were decreased in miR-98-5p-antagomir group(P＜0.05),TUNEL positive rate,the mRNA and protein levels of Bax,IL-1β,IL-6 and TNF-α,STAT3,and the protein level of p-STAT3 and p-STAT3/STAT3 ratio were increased(P＜0.05).②Compared with OA group and NC-agomir+OA group,OARSI score,TUNEL positive rate,the mRNA and protein levels of Bax,IL-1β,IL-6,TNF-α and STAT3 in cartilage tissues,and the protein level of p-STAT3 and p-STAT3/STAT3 ratio in cartilage tissues were decreased in miR-98-5p-agomir+OA group(P＜0.05),while Bcl-2 mRNA and protein level and miR-98-5p level in cartilage tissue were increased(P＜0.05).Conclusion Upregulation of miR-98-5p may inhibit the inflammation and the cell apoptosis in OA by decreasing the activity of STAT3,thus inhibiting the occurrence and development of OA.
osteoarthritismiR-98-5psignal transducer and activator of transcription 3inflammationcell apoptosis
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