Research Paper Volume 13, Issue 16 pp 20258—20276

DNMT3B decreases extracellular matrix degradation and alleviates intervertebral disc degeneration through TRPA1 methylation to inhibit the COX2/YAP axis

DNMT3B alleviated IVDD through TRPA1-COX2-YAP axis. Seven days after IVDD surgery, rats were injected with oe-DNMT3B and/or oe-YAP lentivirus vectors by tail vein for 3 consecutive days. Rats were euthanized 4 weeks after IVDD surgery, and C4-C6 intervertebral disc tissues were collected. (A) DNMT3B, TRPA1, COX2, and YAP expression in IVDD rats treated with oe-DNMT3B and oe-YAP was detected by RT-qPCR. (B) DNMT3B, TRPA1, COX2, and YAP expression in IVDD rats treated with oe-DNMT3B and oe-YAP were detected by Western blot. (C) Histological analysis of the intervertebral disc tissue. (D) The expression of collagen II, aggrecan, MMP3, and MMP9 was detected by RT-qPCR in intervertebral disc tissues of rat models treated with oe-DNMT3B or oe-YAP. (E) The expression of collagen II, aggrecan, MMP3, and MMP9 was detected by Western blot in intervertebral disc tissues of rat models treated with oe-DNMT3B or oe-YAP. (F) Inflammatory factors IL-6, TNF-α, IL-8 levels were detected by ELISA in intervertebral disc tissues of rat models treated with oe-DNMT3B or oe-YAP. Measurement data are expressed as the mean ± standard deviation (n = 3) and analyzed using one-way ANOVA between multiple groups. **, p

Figure 8. DNMT3B alleviated IVDD through TRPA1-COX2-YAP axis. Seven days after IVDD surgery, rats were injected with oe-DNMT3B and/or oe-YAP lentivirus vectors by tail vein for 3 consecutive days. Rats were euthanized 4 weeks after IVDD surgery, and C4-C6 intervertebral disc tissues were collected. (A) DNMT3B, TRPA1, COX2, and YAP expression in IVDD rats treated with oe-DNMT3B and oe-YAP was detected by RT-qPCR. (B) DNMT3B, TRPA1, COX2, and YAP expression in IVDD rats treated with oe-DNMT3B and oe-YAP were detected by Western blot. (C) Histological analysis of the intervertebral disc tissue. (D) The expression of collagen II, aggrecan, MMP3, and MMP9 was detected by RT-qPCR in intervertebral disc tissues of rat models treated with oe-DNMT3B or oe-YAP. (E) The expression of collagen II, aggrecan, MMP3, and MMP9 was detected by Western blot in intervertebral disc tissues of rat models treated with oe-DNMT3B or oe-YAP. (F) Inflammatory factors IL-6, TNF-α, IL-8 levels were detected by ELISA in intervertebral disc tissues of rat models treated with oe-DNMT3B or oe-YAP. Measurement data are expressed as the mean ± standard deviation (n = 3) and analyzed using one-way ANOVA between multiple groups. **, p < 0.01.