Effect of aloperine on chondrocyte injury induced by interleukin-1β and its mechanism / 中国生物制品学杂志

ABSTRACT

@#Objective To investigate the effect of aloperine(ALO)on interleukin-1β(IL-1β)-induced chondrocyte injury and its mechanism. Methods Chondrocytes were randomly divided into control(Con)group,IL-1 β group,IL-1β + ALO-L(25 mg/L)group,IL-1β + ALO-M(50 mg/L)group and IL-1 β + ALO-H(100 mg/L)group;Con group,IL-1βgroup,IL-1β + miR-NC group and IL-1β + miR-16-5p group;Con group,IL-1β group,IL-1β + si-NC group and IL-1β + siSOX5 group. Cells in IL-1β group were treated with 10 ng/mL IL-1β,while no treatment was given in Con group. The transcription levels of miR-16-5p and SOX5 mRNA in chondrocytes were detected by qRT-PCR;The contents of IL-6,TNF-αand IL-1β were detected by ELISA;The expression levels of Bcl-2,Bax and SOX5 protein were detected by Western blot and the apoptosis was detected by flow cytometry. Results Compared with IL-1 β group,the contents of IL-6,TNF-α and IL-1βin IL-1β + ALO-L group,IL-1β + ALO-M group and IL-1β + ALO-H group decreased significantly(t = 5. 002～20. 653,each P < 0. 001),the apoptosis rate decreased significantly(t = 5. 473～17. 371,each P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 7. 800～16. 100,each P < 0. 001),and the expression level of Bax protein decreased significantly(t = 4. 993～14. 311,each P < 0. 001);The mRNA transcription level of miR-16-5p gene increased significantly(t = 6. 688～16. 545,each P < 0. 001),while the mRNA transcription level and protein expression level of SOX5 gene decreased significantly(t = 4. 609～15. 393,each P < 0. 001). Compared with the IL-1β + miR-NC group,the mRNA transcription level of miR-16-5p in the IL-1β + miR-16-5p group increased significantly(t = 17. 106,P < 0. 001),the contents of IL-6,TNF-α and IL-1 β decreased significantly(t = 15. 030～20. 013,each P < 0. 001),the apoptosis rate decreased significantly(t = 12. 273,P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 15. 652,P < 0. 001),and the expression level of Bax protein decreased significantly(t = 12. 999,P < 0. 001). Compared with IL-1β +si-NC group,the expression level of SOX5(t = 13. 444,P < 0. 001),IL-6,TNF-α and IL-1β in IL-1β + si-SOX5 group decreased significantly(t = 14. 087～17. 103,each P < 0. 001),the apoptosis rate decreased significantly(t = 11. 991,P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 13. 864,P < 0. 001),and the expression level of Bax protein decreased significantly(t = 11. 818,P < 0. 001). Conclusion Alo inhibited the apoptosis of chondrocytes induced by IL-1β,thus reducing the injury of chondrocytes,of which the mechanism might be regulating the expression of miR-16-5p and SOX5 and the secretion of inflammatory factors in chondrocytes.