Sirt6促进自噬抑制软骨细胞衰老的体外研究

Abstract
Figure/Table
References (0)
Related Citation (15)

Download: PDF (1957 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Objective: To investigate the effects of Sirt6 in chondrocyte senescence and its effect on chondrocyte autophagy in chondrocyte senescence. Methods: The chondrocytes were isolated and cultured in vitro, and the chondrocyte senescence was induced by continuous passage. The chondrocyte senescence was detected by beta-galactosidase staining, and protein expression of Sirt6 was detected by Western blot. We then overexpressed or knocked down Sirt6 in chondrocytes using lentivirus, and compared the maximum number of passages and the percentages of senescent cells between different groups. The effects of Sirt6 on chondrocyte function were investigated by real-time PCR, to compare the mRNA expression of collagen-II, aggrecan, MMP-13, and ADAMTS-5 in each group. After overexpression of Sirt6 in chondrocytes, the presence of intracellular autophagosomes was analyzed by immunofluorescence, and the ratio of LC3-II/LC3-I was analyzed by Western blot. In addition, the effect of autophagy on the inhibition of senescence and functional degeneration of the chondrocytes during overexpression of Sirt6 was verified by using the autophagy inhibitor, 3-methyladenine. Results: Expression of Sirt6 decreased gradually with an increase in the number of cell generations, and expression of P16 increased significantly in chondrocytes with overexpressed Sirt6. Over expression of Sirt6 in chondrocytes significantly suppressed cellular senescence. Sirt6 promoted the autophagy of chondrocytes, and an autophagy inhibitor inhibited the protective effect of Sirt6 on chondrocytes. Conclusion: Increasing the expression level of Sirt6 can delay the senescence and degeneration of chondrocytes, and its effect is related to the promoted autophagy by Sirt6.

Key words： osteoarthritis chondrocytes cell aging Sirt6 autophagy

Received: 12 September 2017

	Service
	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	ZHANG Hui
	SUN Jiali
	CHEN Zexin
	WU Yaosen
	LIN Yan.

URL:

https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2018/V48/I5/342

[1] SANDELL L J, AIGNER T. Articular cartilage and changes in arthritis. An introduction: cell biology of osteoarthritis[J]. Arthritis Res, 2001, 3(2): 107-113.
[2] RAHMATI M, NALESSO G, MOBASHERI A, et al. Aging and osteoarthritis: central role of the extracellular matrix[J]. Ageing Res Rev, 2017, 40: 20-30.
[3] PIAO J, TSUJI K, OCHI H, et al. Sirt6 regulates postnatal growth plate differentiation and proliferation via Ihh signal-ing[J]. Sci Rep, 2013, 3(10): 3022.
[4] IANNONE F, LAPADULA G. The pathophysiology of os-teoarthritis[J]. Aging Clin Exp Res, 2003, 15(5): 364-372.
[5] 袁普卫, 康武林, 李小群, 等. 骨性关节炎发病机制及相关细胞因子的研究进展[J]. 中国矫形外科杂志, 2016, 24(11): 1010-1015.
[6] PRICE J S, WATERS J G, DARRAH C, et al. The role of chondrocyte senescence in osteoarthritis[J]. Aging Cell, 2002, 1(1): 57-65.
[7] LOESER R F. Age-related changes in the musculoskeletal system and the development of osteoarthritis[J]. Clin Geriatr Med, 2010, 26(3): 371-386.
[8] ZHOU H W, LOU S Q, ZHANG K. Recovery of function in osteoarthritic chondrocytes induced by p16INK4a-specific siRNA in vitro[J]. Rheumatology (Oxford), 2004, 43(5): 555-568.
[9] MOSTOSLAVSKY R, CHUA K F, LOMBARD D B, et al. Genomic instability and aging-like phenotype in the absence of mammalian Sirt6[J]. Cell, 2006, 124(2): 315-329.
[10] KANFI Y, NAIMAN S, AMIR G, et al. The sirtuin Sirt6 regulates lifespan in male mice[J]. Nature, 2012, 483(7388): 218-221.
[11] MICHISHITA E, MCCORD R A, BERBER E, et al. Sirt6 is a histone H3 lysine 9 deacety lase that modulates telomeric chromatin[J]. Nature, 2008, 452(7186): 492-496.
[12] CARDUS A, URYGA A K, WALTERS G, et al. Sirt6 protects human endothelial cells from DNA damage, telomere dysfunction, and senescence[J]. Cardiovasc Res, 2013, 97 (3): 571-579.
[13] BERGAMINI E, CAVALLINI G, DONATI A, et al. The role of autophagy in aging[J]. Ann N Y Acad Sci, 2010, 1114(1): 69-78.
[14] WANG Z J, ZHANG H B, CHEN C, et al. Effect of PPARG on AGEs-induced AKT/MTOR signaling-associated human chondrocytes autophagy[J]. Cell Biol Int, 2018, DOI: 10. 1002/cbin.10951.
[15] DE FIGUEROA P L, LOTZ M K, BLANCO F J, et al.
Autophagy activation and protection from mitochondrial dysfunction in human chondrocytes[J]. Arthritis Rheumatol, 2015, 67(4): 966-976.
[16] 王浩, 曹飞, 斯海波, 等. 雷帕霉素调控自噬在骨关节炎软骨细胞退变中的机制研究[J]. 中华骨与关节外科杂志, 2017, 10(3): 248-253.
[17] CARAMÉS B, HASEGAWA A, TANIGUCHI N, et al. Autophagy activation by rapamycin reduces severity of experimental osteoarthritis[J]. Ann Rheum Dis, 2012, 71(4): 575-581.
[18] MINASHIMA T, ZHANG Y, LEE Y, et al. Lithium protects against cartilage degradation in osteoarthritis[J]. Arthritis Rheumatol, 2014, 66(5): 1228-1236.
[19] SASAKI H, TAKAYAMA K, MATSUSHITA T, et al. Autophagy modulates osteoarthritis-related gene expression in human chondrocytes[J]. Arthritis Rheum, 2012, 64(6): 1920-1928.
[20] MA C H, CHIUA Y C, WU C H, et al. Homocysteine causes dysfunction of chondrocytes and oxidative stress through repression of SIRT1/AMPK pathway: A possible link between hyperhomocysteinemia and osteoarthritis[J]. Redox Biol, 2018, 15: 504-512.
[21] FUJITA N, MATSUSHITA T, ISHIDA K, et al. Potential involvement of SIRT1 in the pathogenesis of osteoarthritis through the modulation of chondrocyte gene expressions[J]. J Orthop Res, 2011, 29(4): 511-515.
[22] MATSUZAKI T, MATSUSHITA T, TAKAYAMA K, et al. Disruption of Sirt1 in chondrocytes causes accelerated progression of osteoarthritis under mechanical stress and during ageing in mice[J]. Ann Rheum Dis, 2014, 73(7): 1397-1404.
[23] GAGARINA V, GABAY O, DVIR-GINZBERG M, et al. Sirt1 enhances survival of human osteoarthritic chondrocytes by repressing protein tyrosine phosphatase 1B and activating the insulin-like growth factor receptor pathway[J]. Arthritis Rheum, 2010, 62(5): 1383-1392.
[24] 李春亮, 李钊伟, 秦凤. Sirt1调控软骨细胞自噬在骨关节炎中作用及机制[J]. 重庆医学, 2016, 45(15): 2118-2122.