Top

European Spine Journal

Published in:

01-10-2018 | Original Article

Lupeol against high-glucose-induced apoptosis via enhancing the anti-oxidative stress in rabbit nucleus pulposus cells

Authors: Ming-Bo Guo, De-Chun Wang, Hai-Fei Liu, Long-Wei Chen, Jian-Wei Wei, Yong Lin, Hui Xue

Published in: European Spine Journal | Issue 10/2018

Abstract

Purpose

This study aimed to investigate the potential mechanism and value of lupeol in inhibiting high-glucose-induced apoptosis in rabbit nucleus pulposus cells (NPCs).

Methods

NPCs were divided into four groups: control (CON), high glucose (HG), LUP, and HG + LUP. Viability, reactive oxygen species (ROS) levels, and apoptosis were examined in NPCs. The protein expression levels of Bax, Bcl-2, cytochrome C, and caspase 9/3 were measured using reverse transcription–polymerase chain reaction and Western blot assay.

Results

The apoptotic rate and total ROS level of the HG group significantly increased compared with the CON group (P < 0.01). The total ROS level in the HG + LUP group significantly decreased compared with the HG group(P < 0.05). The mRNA expression of Bcl-2 was significantly upregulated, whereas the expression of Bax, cytochrome C, and caspase 9/3 was downregulated in the HG + LUP group compared with those in the HG group(P < 0.05).The Western blot assay showed that the expression of Bcl-2 was upregulated, but the expression of Bax, cytochrome C, and caspase 9/3 was significantly downregulated in the HG + LUP group compared with the HG group (P < 0.05).

Conclusions

Lupeol inhibited high-glucose-induced apoptosis in NPCs by enhancing the anti-oxidative stress in the mitochondria. This study suggested lupeol as a potential therapeutic drug for treating intervertebral disc degeneration under hyperglycaemic conditions.

Graphical abstract

These slides can be retrieved under Electronic Supplementary Material.

Available only for authorised users

Sivan SS, Tsitron E, Wachtel E, Roughley PJ, Sakkee N, van der Ham F, DeGroot J, Roberts S, Maroudas A (2006) Aggrecan turnover in human intervertebral disc as determined by the racemization of aspartic acid. J Biol Chem 281:13009–13014. https://doi.org/10.1074/jbc.M600296200 CrossRefPubMed

Samartzis D, Karppinen J, Chan D, Luk KD, Cheung KM (2012) The association of lumbar intervertebral disc degeneration on magnetic resonance imaging with body mass index in overweight and obese adults: a population-based study. Arthritis Rheum 64:1488–1496. https://doi.org/10.1002/art.33462 CrossRefPubMed

Williams FM, Popham M, Sambrook PN, Jones AF, Spector TD, MacGregor AJ (2011) Progression of lumbar disc degeneration over a decade: a heritability study. Ann Rheum Dis 70:1203–1207. https://doi.org/10.1136/ard.2010.146001 CrossRefPubMedPubMedCentral

Potier E, Ito K (2014) Can notochordal cells promote bone marrow stromal cell potential for nucleus pulposus enrichment? A simplified in vitro system. Tissue Eng Part A 20:3241–3251. https://doi.org/10.1089/ten.TEA.2013.0703 CrossRefPubMedPubMedCentral

Battie MC, Videman T, Levalahti E, Gill K, Kaprio J (2008) Genetic and environmental effects on disc degeneration by phenotype and spinal level: a multivariate twin study. Spine 33:2801–2808. https://doi.org/10.1097/BRS.0b013e31818043b7 CrossRefPubMed

Jiang L, Yuan F, Yin X, Dong J (2014) Responses and adaptations of intervertebral disc cells to microenvironmental stress: a possible central role of autophagy in the adaptive mechanism. Connect Tissue Res 55:311–321. https://doi.org/10.3109/03008207.2014.942419 CrossRefPubMed

Won HY, Park JB, Park EY, Riew KD (2009) Effect of hyperglycemia on apoptosis of notochordal cells and intervertebral disc degeneration in diabetic rats. J Neurosurg Spine 11:741–748. https://doi.org/10.3171/2009.6.SPINE09198 CrossRefPubMed

Kong CG, Park JB, Kim MS, Park EY (2014) High glucose accelerates autophagy in adult rat intervertebral disc cells. Asian Spine J 8:543–548. https://doi.org/10.4184/asj.2014.8.5.543 CrossRefPubMedPubMedCentral

Jazini E, Sharan AD, Morse LJ, Dyke JP, Aronowitz EB, Chen LK, Tang SY (2012) Alterations in T2 relaxation magnetic resonance imaging of the ovine intervertebral disc due to nonenzymatic glycation. Spine 37:E209–E215. https://doi.org/10.1097/BRS.0b013e31822ce81f CrossRefPubMedPubMedCentral

10.

Karim L, Tang SY, Sroga GE, Vashishth D (2013) Differences in non-enzymatic glycation and collagen cross-links between human cortical and cancellous bone. Osteoporos Int 24:2441–2447. https://doi.org/10.1007/s00198-013-2319-4 CrossRefPubMedPubMedCentral

11.

Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399. https://doi.org/10.1146/annurev.arplant.55.031903.141701 CrossRefPubMed

12.

Rodriguez R, Redman R (2005) Balancing the generation and elimination of reactive oxygen species. Proc Natl Acad Sci USA 102:3175–3176. https://doi.org/10.1073/pnas.0500367102 CrossRefPubMed

13.

Yan W, Arai A, Aoki M, Ichijo H, Miura O (2007) ASK1 is activated by arsenic trioxide in leukemic cells through accumulation of reactive oxygen species and may play a negative role in induction of apoptosis. Biochem Biophys Res Commun 355:1038–1044. https://doi.org/10.1016/j.bbrc.2007.02.064 CrossRefPubMed

14.

Miyamoto H, Doita M, Nishida K, Yamamoto T, Sumi M, Kurosaka M (2006) Effects of cyclic mechanical stress on the production of inflammatory agents by nucleus pulposus and anulus fibrosus derived cells in vitro. Spine 31:4–9CrossRef

15.

Wang YJ, Shi Q, Lu WW, Cheung KC, Darowish M, Li TF, Dong YF, Zhou CJ, Zhou Q, Hu ZJ, Liu M, Bian Q, Li CG, Luk KD, Leong JC (2006) Cervical intervertebral disc degeneration induced by unbalanced dynamic and static forces: a novel in vivo rat model. Spine 31:1532–1538. https://doi.org/10.1097/01.brs.0000222019.84095.23 CrossRefPubMed

16.

Siddique HR, Saleem M (2011) Beneficial health effects of lupeol triterpene: a review of preclinical studies. Life Sci 88:285–293. https://doi.org/10.1016/j.lfs.2010.11.020 CrossRefPubMed

17.

Lima LM, Perazzo FF, Tavares Carvalho JC, Bastos JK (2007) Anti-inflammatory and analgesic activities of the ethanolic extracts from Zanthoxylum riedelianum (Rutaceae) leaves and stem bark. J Pharm Pharmacol 59:1151–1158. https://doi.org/10.1211/jpp.59.8.0014 CrossRefPubMed

18.

Kumari A, Kakkar P (2012) Lupeol prevents acetaminophen-induced in vivo hepatotoxicity by altering the Bax/Bcl-2 and oxidative stress-mediated mitochondrial signaling cascade. Life Sci 90:561–570. https://doi.org/10.1016/j.lfs.2012.01.012 CrossRefPubMed

19.

Srivastava AK, Mishra S, Ali W, Shukla Y (2016) Protective effects of lupeol against mancozeb-induced genotoxicity in cultured human lymphocytes. Phytomedicine 23:714–724. https://doi.org/10.1016/j.phymed.2016.03.010 CrossRefPubMed

20.

Evens AM, Prachand S, Shi B, Paniaqua M, Gordon LI, Gartenhaus RB (2004) Imexon-induced apoptosis in multiple myeloma tumor cells is caspase-8 dependent. Clin Cancer Res 10:1481–1491CrossRef

21.

Anekstein Y, Smorgick Y, Lotan R, Agar G, Shalmon E, Floman Y, Mirovsky Y (2010) Diabetes mellitus as a risk factor for the development of lumbar spinal stenosis. IMAJ 12:16–20PubMed

22.

Zhang CX, Wang T, Ma JF, Liu Y, Zhou ZG, Wang DC (2017) Protective effect of CDDO-ethyl amide against high-glucose-induced oxidative injury via the Nrf2/HO-1 pathway. Spine J 17:1017–1025. https://doi.org/10.1016/j.spinee.2017.03.015 CrossRefPubMed

23.

Yudoh K, Nguyen VT, Nakamura H, Hongo-Masuko K, Kato T, Nishioka K (2005) Potential involvement of oxidative stress in cartilage senescence and development of osteoarthritis: oxidative stress induces chondrocyte telomere instability and downregulation of chondrocyte function. Arthritis Res Ther 7:R380–R391. https://doi.org/10.1186/ar1499 CrossRefPubMedPubMedCentral

24.

Jallali N, Ridha H, Thrasivoulou C, Underwood C, Butler PE, Cowen T (2005) Vulnerability to ROS-induced cell death in ageing articular cartilage: the role of antioxidant enzyme activity. Osteoarthr Cartil 13:614–622. https://doi.org/10.1016/j.joca.2005.02.011 CrossRefPubMed

25.

Kim KW, Ha KY, Lee JS, Rhyu KW, An HS, Woo YK (2007) The apoptotic effects of oxidative stress and antiapoptotic effects of caspase inhibitors on rat notochordal cells. Spine 32:2443–2448. https://doi.org/10.1097/BRS.0b013e318157395a CrossRefPubMed

26.

Kakkar P, Singh BK (2007) Mitochondria: a hub of redox activities and cellular distress control. Mol Cell Biochem 305:235–253. https://doi.org/10.1007/s11010-007-9520-8 CrossRefPubMed

27.

Singh BK, Tripathi M, Pandey PK, Kakkar P (2010) Nimesulide aggravates redox imbalance and calcium dependent mitochondrial permeability transition leading to dysfunction in vitro. Toxicology 275:1–9. https://doi.org/10.1016/j.tox.2010.05.001 CrossRefPubMed

28.

Heyde CE, Tschoeke SK, Hellmuth M, Hostmann A, Ertel W, Oberholzer A (2006) Trauma induces apoptosis in human thoracolumbar intervertebral discs. BMC Clin Pathol 6:5. https://doi.org/10.1186/1472-6890-6-5 CrossRefPubMedPubMedCentral

29.

Kumari A, Kakkar P (2012) Lupeol protects against acetaminophen-induced oxidative stress and cell death in rat primary hepatocytes. Food Chem Toxicol 50:1781–1789. https://doi.org/10.1016/j.fct.2012.02.042 CrossRefPubMed

30.

Pham TN, Marion M, Denizeau F, Jumarie C (2006) Cadmium-induced apoptosis in rat hepatocytes does not necessarily involve caspase-dependent pathways. Toxicol Vitro 20:1331–1342. https://doi.org/10.1016/j.tiv.2006.05.005 CrossRef

31.

Prasad S, Kalra N, Shukla Y (2008) Induction of apoptosis by lupeol and mango extract in mouse prostate and LNCaP cells. Nutr Cancer 60:120–130. https://doi.org/10.1080/01635580701613772 CrossRefPubMed

32.

Boelsterli UA (2003) Idiosyncratic drug hepatotoxicity revisited: new insights from mechanistic toxicology. Toxicol Mech Methods 13:3–20. https://doi.org/10.1080/15376510309824 CrossRefPubMed

33.

Wuertz K, Urban JP, Klasen J, Ignatius A, Wilke HJ, Claes L, Neidlinger-Wilke C (2007) Influence of extracellular osmolarity and mechanical stimulation on gene expression of intervertebral disc cells. J Orthop Res 25:1513–1522. https://doi.org/10.1002/jor.20436 CrossRefPubMed

34.

Neidlinger-Wilke C, Mietsch A, Rinkler C, Wilke HJ, Ignatius A, Urban J (2012) Interactions of environmental conditions and mechanical loads have influence on matrix turnover by nucleus pulposus cells. J Orthop Res 30:112–121. https://doi.org/10.1002/jor.21481 CrossRefPubMed

35.

Ishihara H, Warensjo K, Roberts S, Urban JP (1997) Proteoglycan synthesis in the intervertebral disk nucleus: the role of extracellular osmolality. Am J Physiol 272:C1499–C1506. https://doi.org/10.1152/ajpcell.1997.272.5.C1499 CrossRefPubMed

36.

Chen J, Baer AE, Paik PY, Yan W, Setton LA (2002) Matrix protein gene expression in intervertebral disc cells subjected to altered osmolarity. Biochem Biophys Res Commun 293:932–938. https://doi.org/10.1016/S0006-291X(02)00314-5 CrossRefPubMed

37.

Mavrogonatou E, Kletsas D (2009) High osmolality activates the G1 and G2 cell cycle checkpoints and affects the DNA integrity of nucleus pulposus intervertebral disc cells triggering an enhanced DNA repair response. DNA Repair 8:930–943. https://doi.org/10.1016/j.dnarep.2009.05.005 CrossRefPubMed

38.

Mavrogonatou E, Papadimitriou K, Urban JP, Papadopoulos V, Kletsas D (2015) Deficiency in the alpha1 subunit of Na+/K+-ATPase enhances the anti-proliferative effect of high osmolality in nucleus pulposus intervertebral disc cells. J Cell Physiol 230:3037–3048. https://doi.org/10.1002/jcp.25040 CrossRefPubMed

39.

Urban JP (2002) The role of the physicochemical environment in determining disc cell behaviour. Biochem Soc Trans 30:858–864. https://doi.org/10.1042/bst0300858 CrossRefPubMed

40.

Hunter CJ, Matyas JR, Duncan NA (2003) The notochordal cell in the nucleus pulposus: a review in the context of tissue engineering. Tissue Eng 9:667–677. https://doi.org/10.1089/107632703768247368 CrossRefPubMed

41.

Hunter CJ, Matyas JR, Duncan NA (2004) Cytomorphology of notochordal and chondrocytic cells from the nucleus pulposus: a species comparison. J Anat 205:357–362. https://doi.org/10.1111/j.0021-8782.2004.00352.x CrossRefPubMedPubMedCentral

Title: Lupeol against high-glucose-induced apoptosis via enhancing the anti-oxidative stress in rabbit nucleus pulposus cells
Authors: Ming-Bo Guo
De-Chun Wang
Hai-Fei Liu
Long-Wei Chen
Jian-Wei Wei
Yong Lin
Hui Xue
Publication date: 01-10-2018
Publisher: Springer Berlin Heidelberg
Published in: European Spine Journal / Issue 10/2018
Print ISSN: 0940-6719
Electronic ISSN: 1432-0932
DOI: https://doi.org/10.1007/s00586-018-5687-9

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Lupeol against high-glucose-induced apoptosis via enhancing the anti-oxidative stress in rabbit nucleus pulposus cells

Abstract

Purpose

Methods

Results

Conclusions

Graphical abstract

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Graphical abstract

Please log in to get access to this content

Other articles of this Issue 10/2018

Correction to: Total disc replacement versus fusion for lumbar degenerative disc disease: a systematic review of overlapping meta-analyses

Does the META score evaluating osteoporotic and metastatic vertebral fractures have enough agreement to be used by orthopaedic surgeons with different levels of training?

TRPC6 in simulated microgravity of intervertebral disc cells

The impact of deep surgical site infection on surgical outcomes after posterior adult spinal deformity surgery: a matched control study

What are the risk factors for surgical site infection after spinal fusion? A meta-analysis

Correction to: Right infraaxillary thoracotomy approach for upper thoracic vertebral decompression and fusion at T2–T6 levels: a technical note