陆军军医大学学报 (Dec 2022)

Kallistatin inhibits apoptosis and extracellular matrix degradation of human nucleus pulposus cells by inducing SIRT1 expression

  • LU Enhu,
  • LI Qingshu,
  • TAN Yun

DOI
https://doi.org/10.16016/j.2097-0927.202205134
Journal volume & issue
Vol. 44, no. 23
pp. 2421 – 2428

Abstract

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Objective To investigate the effect of human tissue kallikrein-binding protein, kallistatin (KS) on intervertebral disc nucleus pulposus cells and the possible mechanism. Methods The expression of KS was detected in normal and degenerative nucleus pulposus cells isolated from the patients undergoing thoracolumbar surgery due to thoracolumbar fracture and lumbar disc herniation in our hospital from April 2020 to August 2021. The cells were divided into Control group, H2O2 group, KS group and H2O2+KS group. The changes of cell apoptosis and extracellular matrix (ECM) in each group were detected. After treatment of silencing information regulator 2 related enzyme Ⅰ (SIRT1) small interfering RNA/KS treatment, the cells were divided into control group, H2O2 group, KS+H2O2 group, KS+H2O2+siRNA-NC group and KS+H2O2+siRNA-SIRT1 group. Cell apoptosis and ECM of the nucleus pulposus cells in each group were detected. Results The mRNA and protein levels of KS were significantly lower in the nucleus pulposus cells isolated from the degenerative intervertebral disc than the normal nucleus pulposus cells (P < 0.05). Compared with the H2O2 group, the apoptotic rate was obviously decreased (P < 0.05), the mRNA and protein levels of apoptosis-related genes Caspase3, Caspase9 and Bax were notably decreased (P < 0.05), the expression of anti-apoptotic gene Bcl2 was remarkably increased, and the content of GAG and the expression levels of Collagen Ⅱ and Aggrecan were statistically increased (P < 0.05) in the KS group and H2O2+KS group (P < 0.05). When the expression of SIRT1 was knocked down by siRNA-SIRT1 in the nucleus pulposus cells, the KS+H2O2+SIRT1 group had significantly increased apoptotic rate, and decreased content of GAG and protein levels of Collagen Ⅱ and Aggrecan when compared with the control group, KS+H2O2 group, and KS+H2O2+siRNA-NC group (P < 0.05). Conclusion KS inhibits H2O2-induced apoptosis and ECM degradation of human nucleus pulposus cells by inducing SIRT1 expression.

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