12 November 2016 : Laboratory Research
17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
Tao WangAC, Si-Dong YangCG, Sen LiuDE, Hui WangCD, Huan LiuCD, Wen Yuan DingCDEFDOI: 10.12659/MSM.900310
Med Sci Monit 2016; 22:4312-4322
Abstract
BACKGROUND: Tumor necrosis factor-α (TNF-α) has been widely known to induce degeneration of nucleus pulposus cells (NPCs). 17β-estradiol (17β-E2) has been broadly proven for its function of suppressing cell apoptosis. The aim of this study is to explore whether 17β-E2 protects apoptosis of human NPCs induced by TNF-α via the PI3K/AKT pathway.
MATERIAL AND METHODS: NPCs were divided into four groups: control, TNF-α (100 ng/mL), TNF-α (100 ng/mL) with pretreated 17β-E2 (10 um/L), TNF-α (100 ng/mL) with pretreated 17β-E2 (10 um/L) and MK2206 (10 um/L, inhibitor of the PI3K/AKT pathway). Flow cytometry was used to measure the apoptotic incidence. Inverted phase-contrast microscopy was used to accomplish the morphological observation for apoptosis of treated cells. Additionally, Cell Counting Kit 8 (CCK-8) assay was used to detected cell proliferation. Western blot and quantitative real-time PCR (qRT-PCR) were applied to explore the expression of pro-caspase-3, caspase-3/p17, cleaved PARP, PARP, Akt, and phospho-Akt (p-Akt).
RESULTS: First, inverted phase-contrast microscopy, CCK-8, and flow cytometry showed that TNF-α induced marked apoptosis, which was abolished by 17β-E2. Furthermore, Western blot and qRT-PCR showed that 17β-E2 protects TNF-α which can induced apoptosis by upregulating p-Akt, whereas Akt was essentially constant. Our data revealed that p-Akt expression peaked at 24 hours in a time-dependent manner (0–48 hours) after treating with TNF-α; and the p-Akt expression generally increased in a time-dependent manner (0–48 hours) after treating with TNF-α and 17β-E2.
CONCLUSIONS: 17β-E2 is shown to protect NPCs against TNF-α induced apoptosis by upregulating p-Akt in the PI3K/AKT pathway. 17β-E2 generally increases expression of p-Akt.
Keywords: Accessory nerve, Estriol, Tumor Necrosis Factors
Errate
The authors asked for the change of the figure 1. They wanted to send the figure that was 200 times bigger under the microscope as described in Figure 1 caption, however, they mistakenly uploaded the wrong picture.
Reference:
1. Tao Wang, Si-Dong Yang, Sen Liu, Hui Wang, Huan Liu, Wen Yuan Ding: 17ß-Estradiol Inhibites Tumor Necrosis Factor-alpha Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway. Med Sci Monit, 2016; 22: 4312-4322. DOI: 10.12659/MSM.900310
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