白果内酯对IL-1β诱导的ATDC5软骨细胞自噬、增殖和凋亡的影响

doi:10.11843/j.issn.0366-6964.2023.02.039

Abstract

Abstract: The purpose of this study was to investigate the effect of bilobalide on IL-1β-induced autophagy, proliferation and apoptosis of ATDC5 chondrocytes. 10 ng·mL^-1 IL-1β was used to induce ATDC5 chondrocytes to construct an in vitro inflammatory model, and then the cells were randomly divided into control group, IL-1β group and bilobalide groups (low-, medium- and high-dosage). The newly synthesized DNA of cells was detected by EdU, and the cell proliferation rate was detected by double labeling combined with the nuclear marker (Hoechest). The apoptosis of ATDC5 chondrocytes was analyzed by flow cytometry using membrane Annexin-V/PI. Autophagic flux was detected by a combined measurement of the mRFP-GFP-LC3 dual fluorescence system. Western blot and qRT-PCR methods were used to detect the protein and mRNA expression levels of LC3-II, Beclin1, BAX, Caspase-3 and Bcl2 in chondrocytes of each group. The results showed that after IL-1β-induced ATDC5 chondrocytes, cell proliferation was attenuated, autophagy was down-regulated and apoptosis was improved. Bilobalide intervention can promote autophagy, proliferation and inhibit apoptosis. Bilobalide promoted the proliferation of IL-1β-induced ATDC5 chondrocytes (P<0.05). Compared with IL-1β group, bilobalide inhibited the protein and mRNA expression of LC3-II, Beclin1, BAX and Caspase-3 (P<0.05), and promoted the protein and mRNA expression of Bcl2 (P<0.05). Autophagy flux was activated, and the apoptosis rate was significantly reduced (P<0.05). The results suggest that bilobalide can promote autophagy of IL-1β-induced ATDC5 chondrocytes and inhibit cell apoptosis, promote cell proliferation and protect chondrocytes.

Key words: articular cartilage, bilobalide, autophagy, apoptosis, cell proliferation, chondrocytes

CLC Number:

S853.74

MA Tianwen, YU Yue, LÜ Liangyu, JIA Lina, RUAN Hongri, WANG Haoran, WANG Xinyu, ZHANG Yuxin, ZHANG Jiantao, GAO Li. Effects of Bilobalide on Autophagy, Proliferation and Apoptosis of IL-1β-induced ATDC5 Chondrocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 837-846.

References 29

[1]	HU X S, ZHANG W Y, LI X, et al.Strategies to modulate the redifferentiation of chondrocytes[J].Front Bioeng Biotechnol, 2021, 9:764193.
[2]	HUNTER D J, BIERMA-ZEINSTRA S.Osteoarthritis[J].Lancet, 2019, 393(10182):1745-1759.
[3]	PÉREZ-LOZANO M L, CESARO A, MAZOR M, et al.Emerging natural-product-based treatments for the management of osteoarthritis[J].Antioxidants (Basel), 2021, 10(2):265.
[4]	SUKHIKH S, NOSKOVA S, IVANOVA S, et al.Chondroprotection and molecular mechanism of action of phytonutraceuticals on osteoarthritis[J].Molecules, 2021, 26(8):2391.
[5]	SIERPINA V S, WOLLSCHLAEGER B, BLUMENTHAL M.Ginkgo biloba[J].Am Fam Physician, 2003, 68(5):923-926.
[6]	LU J, XIE L, LIU K, et al.Bilobalide:A review of its pharmacology, pharmacokinetics, toxicity, and safety[J].Phytother Res, 2021, 35(11):6114-6130.
[7]	MAO D M, LI H, ZHANG L, et al.Bilobalide alleviates IL-17-induced inflammatory injury in ATDC5 cells by downregulation of microRNA-125a[J].J Biochem Mol Toxicol, 2019, 33(12):e22405.
[8]	MA T W, LV L Y, YU Y, et al.Bilobalide exerts anti-inflammatory effects on chondrocytes through the AMPK/SIRT1/mTOR pathway to attenuate ACLT-induced post-traumatic osteoarthritis in rats[J].Front Pharmacol, 2022, 13:783506.
[9]	ELMORE S.Apoptosis:A review of programmed cell death[J].Toxicol Pathol, 2007, 35(4):495-516.
[10]	DUAN R, XIE H, LIU Z Z.The role of autophagy in osteoarthritis[J].Front Cell Dev Biol, 2020, 8:608388.
[11]	CARAMÉS B, OLMER M, KIOSSES W B, et al.The relationship of autophagy defects to cartilage damage during joint aging in a mouse model[J].Arthritis Rheumatol, 2015, 67(6):1568-1576.
[12]	SHUKUNAMI C, SHIGENO C, ATSUMI T, et al.Chondrogenic differentiation of clonal mouse embryonic cell line ATDC5 in vitro:Differentiation-dependent gene expression of parathyroid hormone (PTH)/PTH-related peptide receptor[J].J Cell Biol, 1996, 133(2):457-468.
[13]	HUITEMA L F A, VAN WEEREN P R, BARNEVELD A, et al.Iron ions derived from the nitric oxide donor sodium nitroprusside inhibit mineralization[J].Eur J Pharmacol, 2006, 542(1-3):48-53.
[14]	TIAN Z, ZHANG X N, SUN M L.Phytochemicals mediate autophagy against osteoarthritis by maintaining cartilage homeostasis[J].Front Pharmacol, 2021, 12:795058.
[15]	LI Y L, NIE J L, JIANG P.Oleanolic acid mitigates interleukin-1β-induced chondrocyte dysfunction by regulating miR-148-3p-modulated FGF2 expression[J].J Gene Med, 2020, 22(5):e3169.
[16]	LV S J, WANG X J, JIN S J, et al.Quercetin mediates TSC2-RHEB-mTOR pathway to regulate chondrocytes autophagy in knee osteoarthritis[J].Gene, 2022, 820:146209.
[17]	HU Z C, GONG L F, LI X B, et al.Inhibition of PI3K/Akt/NF-κB signaling with leonurine for ameliorating the progression of osteoarthritis:in vitro and in vivo studies[J].J Cell Physiol, 2019, 234(5):6940-6950.
[18]	JENEI-LANZL Z, MEURER A, ZAUCKE F.Interleukin-1β signaling in osteoarthritis-chondrocytes in focus[J].Cell Signal, 2019, 53:212-223.
[19]	ALMONTE-BECERRIL M, NAVARRO-GARCIA F, GONZALEZ-ROBLES A, et al.Cell death of chondrocytes is a combination between apoptosis and autophagy during the pathogenesis of Osteoarthritis within an experimental model[J].Apoptosis, 2010, 15(5):631-638.
[20]	LIAO M F, LU K T, HSU J L, et al.The role of autophagy and apoptosis in neuropathic pain formation[J].Int J Mol Sci, 2022, 23(5):2685.
[21]	KANG R, ZEH H J, LOTZE M T, et al.The Beclin 1 network regulates autophagy and apoptosis[J].Cell Death Differ, 2011, 18(4):571-580.
[22]	KAO WC, CHEN JC, LIU PC, et al.The Role of Autophagy in Osteoarthritic Cartilage[J].Biomolecules, 2022, 12(10):1357.
[23]	HWANG H S, KIM H A.Chondrocyte apoptosis in the pathogenesis of osteoarthritis[J].Int J Mol Sci, 2015, 16(11):26035-26054.
[24]	王金明, 高宇凤, 成小芳, 等.GSTO1基因沉默对氟致成骨细胞自噬与凋亡的影响[J].畜牧兽医学报, 2019, 50(1):183-192.WANG J M, GAO Y F, CHENG X F, et al.Effects of GSTO1 gene silencing on autophagy and apoptosis of fluoride-induced osteoblasts[J].Acta Veterinaria et Zootechnica Sinica, 2019, 50(1):183-192.(in Chinese).
[25]	EDLICH F. BCL-2 proteins and apoptosis:Recent insights and unknowns[J].Biochem Biophys Res Commun, 2018 27;500(1):26-34.
[26]	ASADI M, TAGHIZADEH S, KAVIANI E, et al. Caspase-3:Structure, function, and biotechnological aspects[J].Biotechnol Appl Biochem, 2022, 69(4).
[27]	XU C, NI S, ZHUANG C, et al.Polysaccharide from Angelica sinensis attenuates SNP-induced apoptosis in osteoarthritis chondrocytes by inducing autophagy via the ERK1/2 pathway[J].Arthritis Res Ther, 2021, 23(1):47.
[28]	DONG J, ZHANG K J, LI G C, et al.CDDO-Im ameliorates osteoarthritis and inhibits chondrocyte apoptosis in mice via enhancing Nrf2-dependent autophagy[J].Acta Pharmacol Sin, 2022, 43(7):1793-1802.
[29]	MAHADEVAN S, PARK Y.Multifaceted therapeutic benefits of Ginkgo biloba L.:chemistry, efficacy, safety, and uses[J].J Food Sci, 2008, 73(1):R14-R19.

Effects of Bilobalide on Autophagy, Proliferation and Apoptosis of IL-1β-induced ATDC5 Chondrocytes

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