Grape seed proanthocyanidins antagonize osteocyte apoptosis due to steroid-induced osteonecrosis of the femoral head

doi:10.3969/j.issn.2095-4344.2759

Abstract

Abstract:

BACKGROUND: Steroid-induced avascular necrosis of the femoral head has a complex biological process, and its pathogenesis is unknown. To date, there is no effective treatment in clinical practice. Therefore, exploring the etiology of steroid-induced avascular necrosis of the femoral head is still an important content of research in this field.

OBJECTIVE: To explore the effect of grape seed proanthocyanidins on osteocyte apoptosis due to steroid-induced osteonecrosis of the femoral head.

METHODS: Twenty-seven Japanese white rabbits were randomly divided into three groups. The model group was intravenously injected with E. coli endotoxin, 100 μg/kg, twice at an interval of 24 hours. Two injections of E. coli endotoxin were followed by intramuscular injection of methylprednisolone 20 mg/kg, for 3 times. The interval was 24 hours. The treatment group was intravenously injected with E. coli endotoxin, 100 μg/kg, twice at an interval of 24 hours. After the second injection of E. coli endotoxin, methylprednisolone 20 mg/kg and grape seed proanthocyanidin extract 200 μg/kg were injected intramuscularly three times at an interval of 24 hours. The control group was treated with the same dose of saline intravenously. The animals were killed by air embolization at the 4^th week after the last injection. Under the relatively aseptic condition, the bilateral femoral heads were routinely fixed, decalcified, embedded and sliced. Histomorphological observation and hematoxylin-eosin staining were performed to count empty bone lacunae under light microscope. Hoechst staining was used to detect cell apoptosis. The expressions of Caspase-9 and Bcl-2 in the femoral head were detected by immunohistochemical staining.

RESULTS AND CONCLUSION: The percentage of empty bone lacunae in the model group was significantly higher than that in the control group (P < 0.01). The percentage of empty bone lacunae in the treatment group was significantly lower than that in the model group (P < 0.05), but still higher than that in the control group (P < 0.01). There were few apoptotic cells in the femoral head of the control group, but a large number of apoptotic cells with densely stained nuclei in the model group. The number of apoptotic cells in the treatment group was reduced, but still higher than that in the control group (P < 0.01). The average absorbance value of Caspase-9 in the model group was significantly higher than that in the control group (P < 0.01). The average absorbance value decreased significantly in the treatment group as compared with the model group (P < 0.01), but there was no significant difference between the treatment and control groups. The average absorbance value of Bcl-2 in the model group was significantly lower than that in the control group (P < 0.05). The average absorbance value of Bcl-2 increased significantly in the treatment group as compared with the model group (P < 0.01). These findings indicate that osteocyte apoptosis is involved in the occurrence and development of steroid-induced osteonecrosis of the femoral head. Grape seed proanthocyanidins may inhibit osteocyte apoptosis by promoting the expression of Bcl-2 and inhibiting the expression of Caspase-9.

Key words: glucocorticoid, osteonecrosis of the femoral head, apoptosis, grape seed proanthocyanidins, Caspase-9, Bcl-2

CLC Number:

Zhang Zhifeng, Wei Jing, Sun Tao, Song Jinling, Zhao Zhenqun, Huang Jian. Grape seed proanthocyanidins antagonize osteocyte apoptosis due to steroid-induced osteonecrosis of the femoral head[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(26): 4146-4151.

Figures/Tables 6

References

[1] DRESCHER W, SCHLIEPER G, FLOEGE J, et al. Steroid-related osteonecrosis--an update. Nephrol Dial Transplant. 2011;26(9): 2728-2731.

[2] HWANG Y, PARK J, CHOI SH, et al. Traumatic and Non-traumatic Osteonecrosis in the Femoral Head of a Rabbit Model. Lab Anim Res. 2011;27(2):127-131.

[3] HU X, GUO Z, XU J, et al. Role of feline sarcoma‑related protein in the viability and apoptosis of bladder cancer cells. Mol Med Rep. 2019;19(6): 5219-5226.

[4] HUANG J, DING Z, LUO Q, et al. Cancer cell-derived exosomes promote cell proliferation and inhibit cell apoptosis of both normal lung fibroblasts and non-small cell lung cancer cell through delivering alpha-smooth muscle actin. Am J Transl Res. 2019;11(3):1711-1723.

[5] CAO M, ZHANG Z, HAN S, et al. Butyrate inhibits the proliferation and induces the apoptosis of colorectal cancer HCT116 cells via the deactivation of mTOR/S6K1 signaling mediated partly by SIRT1 downregulation. Mol Med Rep. 2019;19(5):3941-3947.

[6] KYLLI P, NOHYNEK L, PUUPPONEN-PIMIÄ R, et al. Lingonberry (Vaccinium vitis-idaea) and European cranberry (Vaccinium microcarpon) proanthocyanidins: isolation, identification, and bioactivities. J Agric Food Chem. 2011;59(7):3373-3384.

[7] 范明远,叶青.体内自由基清除剂及抗氧化剂—原花青素的研究进展[J].中国预防医学杂志2001,12(4):303-305.

[8] ZHANG Z, ZHENG L, ZHAO Z, et al. Grape seed proanthocyanidins inhibit H2O2-induced osteoblastic MC3T3-E1 cell apoptosis via ameliorating H2O2-induced mitochondrial dysfunction. J Toxicol Sci. 2014;39(5):803-813.

[9] ZHAO ZQ, BAI R, LIU WL, et al. Roles of oxidative DNA damage of bone marrow hematopoietic cells in steroid-induced avascular necrosis of femoral head. Genet Mol Res. 2016;15(1):15017706.

[10] PAPAVASILIOU AV, TRIANTAFYLLOPOULOS I, PAXINOS O, et al. The role of cell therapies and hip arthroscopy in the management of osteonecrosis: an update. J Hip Preserv Surg. 2018;5(3):202-208.

[11] MCAVOY S, BAKER KS, MULROONEY D, et al. Corticosteroid dose as a risk factor for avascular necrosis of the bone after hematopoietic cell transplantation. Biol Blood Marrow Transplant. 2010;16(9):1231-1236.

[12] YAMAMOTO T, SCHNEIDER R, IWAMOTO Y, et al. Rapid destruction of the femoral head after a single intraarticular injection of corticosteroid into the hip joint. J Rheumatol. 2006;33(8):1701-1704.

[13] WEINSTEIN RS, JILKA RL, PARFITT AM, et al. Inhibition of osteoblastogenesis and promotion of apoptosis of osteoblasts and osteocytes by glucocorticoids. Potential mechanisms of their deleterious effects on bone. J Clin Invest. 1998;102(2):274-282.

[14] O'BRIEN CA, JIA D, PLOTKIN LI, et al. Glucocorticoids act directly on osteoblasts and osteocytes to induce their apoptosis and reduce bone formation and strength. Endocrinology. 2004;145(4):1835-1841.

[15] KERACHIAN MA, HARVEY EJ, COURNOYER D, et al. A rat model of early stage osteonecrosis induced by glucocorticoids. J Orthop Surg Res. 2011;6:62.

[16] WEINSTEIN RS, WAN C, LIU Q, et al. Endogenous glucocorticoids decrease skeletal angiogenesis, vascularity, hydration, and strength in aged mice. Aging Cell. 2010;9(2):147-161.

[17] KERACHIAN MA, SÉGUIN C, HARVEY EJ. Glucocorticoids in osteonecrosis of the femoral head: a new understanding of the mechanisms of action. J Steroid Biochem Mol Biol. 2009;114(3-5): 121-128.

[18] WEINSTEIN RS, NICHOLAS RW, MANOLAGAS SC. Apoptosis of osteocytes in glucocorticoid-induced osteonecrosis of the hip. J Clin Endocrinol Metab. 2000;85(8):2907-2912.

[19] KABATA T, KUBO T, MATSUMOTO T, et al. Apoptotic cell death in steroid induced osteonecrosis: an experimental study in rabbits. J Rheumatol. 2000;27(9):2166-2171.

[20] NIE Z, CHEN S, PENG H. Glucocorticoid induces osteonecrosis of the femoral head in rats through GSK3β-mediated osteoblast apoptosis. Biochem Biophys Res Commun. 2019;511(3):693-699.

[21] CHEN Y, DOU C, YI J, et al. Inhibitory effect of vanillin on RANKL-induced osteoclast formation and function through activating mitochondrial-dependent apoptosis signaling pathway. Life Sci. 2018; 208:305-314.

[22] JIN YR, STOHN JP, WANG Q, et al. Inhibition of osteoclast differentiation and collagen antibody-induced arthritis by CTHRC1. Bone. 2017;97: 153-167.

[23] CONAWAY HH, HENNING P, LIE A, et al. Activation of dimeric glucocorticoid receptors in osteoclast progenitors potentiates RANKL induced mature osteoclast bone resorbing activity. Bone. 2016;93:43-54.

[24] LAI R, XIAN D, XIONG X, et al. Proanthocyanidins: novel treatment for psoriasis that reduces oxidative stress and modulates Th17 and Treg cells. Redox Rep. 2018;23(1):130-135.

[25] ZHAO YM, GAO LP, ZHANG HL, et al. Grape seed proanthocyanidin extract prevents DDP-induced testicular toxicity in rats. Food Funct. 2014;5(3):605-611.

[26] YANG D, LI S, GAO L, et al. Dietary grape seed procyanidin extract protects against lead-induced heart injury in rats involving endoplasmic reticulum stress inhibition and AKT activation. J Nutr Biochem. 2018;62: 43-49.

[27] NIU Q, HE P, XU S, et al. Fluoride-induced iron overload contributes to hepatic oxidative damage in mouse and the protective role of Grape seed proanthocyanidin extract. J Toxicol Sci. 2018;43(5):311-319.