Polydatin effects on apoptosis of nucleus pulposus cells and SIRT1/mTOR pathway in mice with lumbar disc degeneration

doi:10.12307/2021.290

Abstract

Abstract: BACKGROUND: Intervertebral disc degeneration often causes low back pain, shoulder and neck pain in the elderly, which is closely related to the apoptosis, inflammation and oxidative stress of nucleus pulposus cells in intervertebral disc degeneration. Polydatin is the main active component of Polygonum cuspidatum, which has anti-inflammatory, antioxidant and other biological activities.
OBJECTIVE: To investigate the effects of polydatin on apoptosis of nucleus pulposus cells and silent information regulator 1 (SIRT1)/mammalian target of rapamycin (mTOR) pathway in mice with intervertebral disc degeneration.
METHODS: Primary mouse nucleus pulposus cells were isolated and cultured by trypsin. Cell identification and cytotoxicity measurement were conducted by toluidine blue staining and type II collagen immunocytochemistry staining. The nucleus pulposus cells were randomly divided into five groups: blank group, model group, low-, middle-, and high-dose polydatin groups (10, 25, and 50 mg/L). The blank group was normally cultured; in addition, the other groups were induced by interleukin-1β to build the degenerative model of nucleus pulposus cells, followed by cultured with different doses of polydatin. Cell apoptosis in each group was detected by flow cytometry. The expression levels of oxidative stress factors and inflammatory factors were measured by enzyme-linked immunosorbent assay. The expression levels of apoptosis related proteins and SIRT1/mTOR signaling pathway related proteins in nucleus pulposus cells were detected by western blot method. The study protocol was ethically approved by the Ethics Committee of General Hospital of the Yangtze River Shipping with an approval No. L20200059.
RESULTS AND CONCLUSION: The nucleus pulposus cells were successfully isolated and purified by trypsin. Compared with those in the blank group, the apoptotic rate of nucleus pulposus cells, the expression of Caspase-3 and Bax proteins, the expression levels of malondialdehyde, lactate dehydrogenase, interleukin-6, inducible nitric oxide synthase, and tumor necrosis factor α were significantly higher in the model group. Compared with the model group, these indexes were significantly decreased in the polydatin groups. Compared with the blank group, the model group showed a significant reduction in the expression levels of Bcl-2 protein, superoxide dismutase, reduced glutathione, SIRT1, phosphorylated PI3K/phosphatidylinositol-3-kinase, phosphorylated Akt/protein kinase B and mTOR protein (P < 0.05), and the polydatin groups had a significant increase in the above-mentioned indexes in a dose-dependent manner (P < 0.05). To conclude, polydatin can significantly reduce the apoptosis, inflammation and oxidative stress of nucleus pulposus cells induced by interleukin-1β, which may be achieved by activating the SIRT1/mTOR pathway.

Key words: polydatin, lumbar disc degeneration, nucleus pulposus cells, silent information regulator 1, mammalian target of rapamycin

CLC Number:

Zuo Bin, Xia Xiaofeng, Che Biao, Zou Kai, Tang Jiaguo. Polydatin effects on apoptosis of nucleus pulposus cells and SIRT1/mTOR pathway in mice with lumbar disc degeneration[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(35): 5619-5625.

Figures/Tables 9

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