Abstract
Objective To investigate the effect of small interfering RNA targeting mechanosensitive ion channel protein Piezo1 (Piezo1-siRNA) on abnormal chondrocyte proliferation exposed to mechanical stretch. Methods Construct and screen effective Piezo1-siRNA sequences and explore an appropriate method to transfect lentiviral vector into chondrocytes exposed to mechanical stretch. Western blot and RT-PCR were used to detect the mRNA and protein expression of Piezo1, Kif18A, and β-tubulin, respectively. Flow cytometry was used to measure the changes in the chondrocyte cycle. The proliferation of chondrocyte was evaluated by cell counting kit-8. Results According to the mRNA and protein expression of Piezo1, the effective siRNA sequence was successfully screened. Compared with the 0 h group, mechanical stretch upregulated the expression of Piezo1, Kif18A, and β-tubulin, resulting in chondrocyte cycle arrest and eventually inhibiting chondrocyte proliferation. Moreover, Piezo1-siRNA transfection effectively blocks this process and promotes the proliferation of chondrocyte. Conclusion Piezo1-siRNA can reduce the inhibition of chondrocyte proliferation induced by mechanical stretch via downregulating the expression of Kif18A and inhibiting the depolymerization of microtubules. Piezo1-siRNA plays a protective role in chondrocytes, which provides a potential method for the treatment of OA under abnormal mechanical stimulation.
Highlights
Osteoarthritis (OA) is a degenerative disease of bone and articular cartilage, which is caused by mechanical and biological factors [1,2,3]
Previous studies have reported that the expression of Piezo1 in chondrocyte exposed to different mechanical stretch is various, and chondrocyte proliferation is closely associated with the time of mechanical stretch [11]
The expression of Piezo1 in chondrocytes changed with the extension of mechanical stretch time, and the expression of Piezo1 protein was significantly inhibited by transfection of siRNA
Summary
Osteoarthritis (OA) is a degenerative disease of bone and articular cartilage, which is caused by mechanical and biological factors [1,2,3]. Degenerative changes of articular cartilage are closely related to abnormal proliferation of chondrocyte [4, 5], in which mechanical stretch stress plays a crucial role [6]. Piezo is a novel type of nonselective mechanically sensitive ion channel which is different from other ion channels in structure and gating mechanism [8, 9] It converts mechanical signals into chemical signals and engages in cell proliferation, apoptosis, and metabolism [10]. Previous studies have reported that the expression of Piezo in chondrocyte exposed to different mechanical stretch is various, and chondrocyte proliferation is closely associated with the time of mechanical stretch [11]. Whether Piezo is involved in mechanical stretch-induced abnormal proliferation of chondrocytes is still unclear
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