Resveratrol Combined with 17<i>β</i>-Estradiol Prevents IL-1<i>β</i> Induced Apoptosis in Human Nucleus Pulposus Via The PI3K/AKT/Mtor and PI3K/AKT/GSK-3<i>β</i> Pathway
Xiaoliang Bai, Xiaohui Guo, Feng Zhang, Long Tai Zheng, Wenyuan Ding, Sidong Yang
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.