Resveratrol Combined with 17β-Estradiol Prevents IL-1β Induced Apoptosis in Human Nucleus Pulposus Via The PI3K/AKT/Mtor and PI3K/AKT/GSK-3β Pathway

Abstract

Backgrounds Nucleus pulposus (NP) apoptosis is mainly charged for the pathological process of Intervertebral disc degeneration (IVDD). Our previous study revealed that Resveratrol (RSV) combined with 17β-estradiol (E2) was more effective in cutting down IL-1β induced NP cell apoptosis via PI3K/AKT pathway. The present study further evaluated the effect of RSV and E2 in the anti-apoptosis process of IVDD. Methods Human nucleus pulposus (NP) cells culture system and IL-1β inducing apoptosis model were constructed in this research. RSV and E2 were used to inhibit apoptosis. FACS (Fluorescence-activated cell sorting) and CCK-8 (Cell Counting Kit-8) assays were respectively used to determine apoptotic incidence and cell viability of NP cells. Quantitative RT-PCR was used to determine expression of target genes in mRNA level, and western blot analysis was performed to detect the changes of related protein expression. Results RSV combined with E2 attenuated IL-1β-induced cell apoptosis and recovered cell viability. Blockers for mTOR and GSK-3β abated the effect of RSV and E2. RSV combined with E2 obviously increased activated P-mTOR and P-GSK-3β, which contributes to the downregulation of caspase-3. Activated P-NF-kappa B was not involved in the anti-apoptosis process of RSV and E2. Conclusion Combination of Resveratrol and 17β-estradiol efficiently resisted IL-1β induced apoptosis of NP cell, mainly through PI3K/AKT/mTOR/caspase-3 and PI3K/AKT/GSK-3β pathway.

Keywords

• nucleus pulposus (np)
• intervertebral disc degeneration (ivdd)
• resveratrol
• 17β-estradiol