Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy
Background/Aims: Intervertebral discs consist of an extracellular matrix (ECM) with a central gelatinous nucleus pulposus (NP) enclosed in an outer layer known as the annulus fibrosus. ECM metabolic disorders result in loss of boundary between the annulus fibrosus and NP, which can lead to intervert...
| Main Authors: | , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Cell Physiol Biochem Press GmbH & Co KG
2018-09-01
|
| Series: | Cellular Physiology and Biochemistry |
| Subjects: | |
| Online Access: | https://www.karger.com/Article/FullText/493843 |
| _version_ | 1819068038540951552 |
|---|---|
| author | Changgui Shi Huiqiao Wu Di Du Hee-Jeong Im Ying Zhang Bo Hu Huajiang Chen Xinwei Wang Yang Liu Peng Cao Ye Tian Xiaolong Shen Rui Gao Andre J van Wijnen Xiaojian Ye Wen Yuan |
| author_facet | Changgui Shi Huiqiao Wu Di Du Hee-Jeong Im Ying Zhang Bo Hu Huajiang Chen Xinwei Wang Yang Liu Peng Cao Ye Tian Xiaolong Shen Rui Gao Andre J van Wijnen Xiaojian Ye Wen Yuan |
| author_sort | Changgui Shi |
| collection | DOAJ |
| description | Background/Aims: Intervertebral discs consist of an extracellular matrix (ECM) with a central gelatinous nucleus pulposus (NP) enclosed in an outer layer known as the annulus fibrosus. ECM metabolic disorders result in loss of boundary between the annulus fibrosus and NP, which can lead to intervertebral disc degeneration (IDD). Proinflammatory cytokines, such as interleukin (IL)-1β, mediate the progression of IDD. Nicotinamide phosphoribosyltransferase (Nampt) catalyzes the first step in the biosynthesis of nicotinamide adenine dinucleotide (NAD) and is known to be induced by IL-1β. APO866 is an inhibitor of NAD biosynthesis and is involved in autophagy. LC3 (microtubule-associated protein 1 light chain 3) is a key regulator of autophagy and is used as an indicator of increased autophagy. Herein, we investigate the role of APO866 in regulating autophagy in NP cells and IL-1β mediated NP cell degeneration and apoptosis. Methods: NP cells were extracted from IDD tissues and cultured in DMEM/F12 medium. Nampt was induced by different concentrations of IL-1β (0, 0.5, 1, 5, 10 ng/mL) for 24 h or NP cells were treated with 10 ng/mL IL-1β for 0, 6, 12, 48 h. QRT-PCR and western blots were used to detect Nampt and ECM-related protein expression in NP tissue of patients with IDD and in NP cells. Confocal analysis was used to detect membrane-bound LC3, Aggrecan, and Collagen II. Results: Nampt is expressed in NP tissue at higher levels in severe grades of IDD (Grade IV and V) compared with low grades (Grade II and III). In NP cells, 10 ng/mL IL-1β induced Nampt expression for 48 h, increased expression of the degradative-associated proteins, ADAMTS4/5 and MMP-3/13, and decreased expression of ECM-related proteins, Aggrecan and Collagen II. However, the Nampt inhibitor APO866 blocked IL-1β induction, and the knockdown of Nampt expression increased the expression of ECM proteins that were inhibited by IL-1β. Moreover, evidence provided by the autophagic markers LC3 and Beclin-1 indicated that APO866 induced NP cell autophagy. Furthermore, although APO866 inhibited the downregulated expression of ECM-related proteins by IL-1β, this function was blocked by autophagy inhibitor, 3-methyladenine. Conclusion: APO866 protects NP cells and induces autophagy by inhibiting IL-1β-induced NP cell degeneration and apoptosis, which may have therapeutic potential in IDD. |
| first_indexed | 2024-12-21T16:27:48Z |
| format | Article |
| id | doaj.art-e11f0ffdb3154686ba67748571a8f89b |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-21T16:27:48Z |
| publishDate | 2018-09-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-e11f0ffdb3154686ba67748571a8f89b2022-12-21T18:57:25ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-09-014962463248210.1159/000493843493843Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via AutophagyChanggui ShiHuiqiao WuDi DuHee-Jeong ImYing ZhangBo HuHuajiang ChenXinwei WangYang LiuPeng CaoYe TianXiaolong ShenRui GaoAndre J van WijnenXiaojian YeWen YuanBackground/Aims: Intervertebral discs consist of an extracellular matrix (ECM) with a central gelatinous nucleus pulposus (NP) enclosed in an outer layer known as the annulus fibrosus. ECM metabolic disorders result in loss of boundary between the annulus fibrosus and NP, which can lead to intervertebral disc degeneration (IDD). Proinflammatory cytokines, such as interleukin (IL)-1β, mediate the progression of IDD. Nicotinamide phosphoribosyltransferase (Nampt) catalyzes the first step in the biosynthesis of nicotinamide adenine dinucleotide (NAD) and is known to be induced by IL-1β. APO866 is an inhibitor of NAD biosynthesis and is involved in autophagy. LC3 (microtubule-associated protein 1 light chain 3) is a key regulator of autophagy and is used as an indicator of increased autophagy. Herein, we investigate the role of APO866 in regulating autophagy in NP cells and IL-1β mediated NP cell degeneration and apoptosis. Methods: NP cells were extracted from IDD tissues and cultured in DMEM/F12 medium. Nampt was induced by different concentrations of IL-1β (0, 0.5, 1, 5, 10 ng/mL) for 24 h or NP cells were treated with 10 ng/mL IL-1β for 0, 6, 12, 48 h. QRT-PCR and western blots were used to detect Nampt and ECM-related protein expression in NP tissue of patients with IDD and in NP cells. Confocal analysis was used to detect membrane-bound LC3, Aggrecan, and Collagen II. Results: Nampt is expressed in NP tissue at higher levels in severe grades of IDD (Grade IV and V) compared with low grades (Grade II and III). In NP cells, 10 ng/mL IL-1β induced Nampt expression for 48 h, increased expression of the degradative-associated proteins, ADAMTS4/5 and MMP-3/13, and decreased expression of ECM-related proteins, Aggrecan and Collagen II. However, the Nampt inhibitor APO866 blocked IL-1β induction, and the knockdown of Nampt expression increased the expression of ECM proteins that were inhibited by IL-1β. Moreover, evidence provided by the autophagic markers LC3 and Beclin-1 indicated that APO866 induced NP cell autophagy. Furthermore, although APO866 inhibited the downregulated expression of ECM-related proteins by IL-1β, this function was blocked by autophagy inhibitor, 3-methyladenine. Conclusion: APO866 protects NP cells and induces autophagy by inhibiting IL-1β-induced NP cell degeneration and apoptosis, which may have therapeutic potential in IDD.https://www.karger.com/Article/FullText/493843Nucleus pulposusIL-1βNamptAPO866ECMAutophagy |
| spellingShingle | Changgui Shi Huiqiao Wu Di Du Hee-Jeong Im Ying Zhang Bo Hu Huajiang Chen Xinwei Wang Yang Liu Peng Cao Ye Tian Xiaolong Shen Rui Gao Andre J van Wijnen Xiaojian Ye Wen Yuan Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy Cellular Physiology and Biochemistry Nucleus pulposus IL-1β Nampt APO866 ECM Autophagy |
| title | Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy |
| title_full | Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy |
| title_fullStr | Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy |
| title_full_unstemmed | Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy |
| title_short | Nicotinamide Phosphoribosyltransferase Inhibitor APO866 Prevents IL-1β-Induced Human Nucleus Pulposus Cell Degeneration via Autophagy |
| title_sort | nicotinamide phosphoribosyltransferase inhibitor apo866 prevents il 1β induced human nucleus pulposus cell degeneration via autophagy |
| topic | Nucleus pulposus IL-1β Nampt APO866 ECM Autophagy |
| url | https://www.karger.com/Article/FullText/493843 |
| work_keys_str_mv | AT changguishi nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT huiqiaowu nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT didu nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT heejeongim nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT yingzhang nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT bohu nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT huajiangchen nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT xinweiwang nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT yangliu nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT pengcao nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT yetian nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT xiaolongshen nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT ruigao nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT andrejvanwijnen nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT xiaojianye nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy AT wenyuan nicotinamidephosphoribosyltransferaseinhibitorapo866preventsil1binducedhumannucleuspulposuscelldegenerationviaautophagy |