Open AccessHydrogen sulfide is expressed in the human and the rat cultured nucleus pulposus cells and suppresses apoptosis induced by hypoxia
Author(s) -
Haolin Sun,
Litong Qi,
Shijun Wang,
Xuwen Li,
Chunde Li
Publication year - 2018
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0192556
Subject(s) - apoptosis , cystathionine beta synthase , hypoxia (environmental) , annexin , tunel assay , microbiology and biotechnology , endogeny , programmed cell death , cell culture , chemistry , biology , biochemistry , oxygen , enzyme , cysteine , organic chemistry , genetics
Apoptosis plays pivotal role in the pathogenesis of degenerative disc diseases, which is the primary contributor to low back pain. Although the role of hydrogen sulfide (H 2 S) in cell apoptosis is well appreciated, the effects and mechanism that H 2 S regulates the program death of intervertebral disc cell are not yet elucidated. In this study, we utilized the nucleus pulposus (NP) from patients with lumbar disc herniation to investigate the relationship between endogenous H 2 S and NP cells apoptosis in human. Furthermore, we analyzed primary rat NP cells to study the effects of exogenous H 2 S on hypoxia induced cell apoptosis. Human NP samples were obtained from patients with lumbar disc herniation and were divided into uncontained and contained herniation groups. Using immunohistochemistry staining and sulphur-sensitive electrode, we detected the expression of cystathionine-β-synthase (CBS) and cystathionine γ-lyase (CSE), as well as the production of endogenous H 2 S in human NP. Tunel staining showed increased apoptosis in NP from herniated disc; and there was significant correlation between H 2 S generation and apoptosis in human NP. CoCl 2 was then used to induce hypoxia in cultured primary rat NP cells. Annexin V staining indicated that exogenous NaHS attenuated hypoxia induced apoptosis in rat NP cells. Furthermore, hypoxia significantly increased the levels of multiple apoptosis associated proteins (Fas, Cytochromes C, Caspase 9 and cleaved-Caspase-3) in cells, which were eliminated by NaHS. Our study demonstrates the presence of endogenous H 2 S in human intervertebral disc; and the endogenous H 2 S generation rate is associated with NP apoptosis in herniated disc. In vitro study showes exogenous H 2 S donor attenuates hypoxia induced apoptosis in primary rat NP cells. Thus, our work provides insights that H 2 S may have beneficial effects in treating degenerative disc diseases.