预防性使用多孔纳米硒材料Se@SiO_2缓解激素性股骨头坏死的实验研究
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摘要
[目的]探究预防性使用多孔纳米硒材料Se@SiO_2能否抑制激素性股骨头坏死(SANFH)的发生。[方法]体外实验:选取SPF级4周龄SD大鼠2只,提取、培养并鉴定股骨头软骨细胞,利用细胞增殖与活性检测试剂(CCK-8)检测Se@SiO_2在应用浓度下对软骨细胞活性的影响,再利用活性氧试剂盒检测Se@SiO_2对软骨细胞抗氧化应激作用的影响。体内实验:选取SPF级8周龄SD大鼠24只,随机选取12只SD大鼠作为实验组,每2 d腹腔注射1mg/kg Se@SiO_2共2周,之后24只大鼠利用静脉给予甲强龙建立激素性股骨头坏死动物模型,通过股骨头软骨组织IL-1β、MMP-13的表达和HE染色切片评估预防性使用Se@SiO_2对股骨头软骨组织坏死的改善作用。[结果](1)Se@SiO_2在应用浓度下对股骨头软骨细胞无明显细胞毒性,并且加入Se@SiO_2干预可有效清除软骨细胞活性氧水平;(2)提前加入Se@SiO_2干预2周后股骨头软骨组织IL-1β、MMP13表达明显下降,HE染色显示提前使用Se@SiO_2干预能有效改善SANFH程度。[结论]提前2周给予Se@SiO_2,能改善SANFH程度。
[Objective] To explore whether prophylactic use of porous nano selenium material Se@SiO_2 inhibit the occurrence of steroid-induced avascular necrosis of femoral head(SANFH). [Method] Regarding to the experiment in vitro, the chondrocytes, extracted from femoral heads of 2 specific pathogen free(SPF) SD rats aged 4 weeks, were incubated and identified. The CCK-8 assay was applied to detect the impact of Se@SiO_2 in the applied concentration on cell proliferation and activity, while the reactive oxygen species assay was used to detect effect of Se@SiO_2 in the applied concentration on the oxidative stress of the femoral head chondrocytes. Regarding to the experiment in vivo, 24 SPF SD rats aged 8 weeks were randomly divided into the experimental group and the control group with 12 animals in each group, the animals in experimental group were intraperitoneally given Se@SiO_21 mg/kg every two days for 2 weeks. After that, all the 24 rats were intravenously given methylprednisolone(MPS) to establish the SANFH models. The expression IL-1 beta and MMP-13 detected by Western-Blot assay,as well as histological observations were conducted to compare the two groups. [Results] In vitro, the Se@SiO_2 had no obvious cytotoxicity on the chondrocytes of femoral head under the applied concentration, whereas decreased of the oxidative stress of the femoral head chondrocyte. In vivo, the Se@SiO_2 reduced IL-1β and MMP13 expression in the femoral head, improved histological appearance of SANFH. [Conclusion] Application of Se@SiO_2 two weeks advance does alleviate pathogenesis of SANFH.
[Objective] To explore whether prophylactic use of porous nano selenium material Se@SiO_2 inhibit the occurrence of steroid-induced avascular necrosis of femoral head(SANFH). [Method] Regarding to the experiment in vitro, the chondrocytes, extracted from femoral heads of 2 specific pathogen free(SPF) SD rats aged 4 weeks, were incubated and identified. The CCK-8 assay was applied to detect the impact of Se@SiO_2 in the applied concentration on cell proliferation and activity, while the reactive oxygen species assay was used to detect effect of Se@SiO_2 in the applied concentration on the oxidative stress of the femoral head chondrocytes. Regarding to the experiment in vivo, 24 SPF SD rats aged 8 weeks were randomly divided into the experimental group and the control group with 12 animals in each group, the animals in experimental group were intraperitoneally given Se@SiO_21 mg/kg every two days for 2 weeks. After that, all the 24 rats were intravenously given methylprednisolone(MPS) to establish the SANFH models. The expression IL-1 beta and MMP-13 detected by Western-Blot assay,as well as histological observations were conducted to compare the two groups. [Results] In vitro, the Se@SiO_2 had no obvious cytotoxicity on the chondrocytes of femoral head under the applied concentration, whereas decreased of the oxidative stress of the femoral head chondrocyte. In vivo, the Se@SiO_2 reduced IL-1β and MMP13 expression in the femoral head, improved histological appearance of SANFH. [Conclusion] Application of Se@SiO_2 two weeks advance does alleviate pathogenesis of SANFH.
引文
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[18]Komurcu E.Preventive effects of coenzyme Q10(Co Q10)on steroid-induced osteonecrosis in rats[J].Acta Orthop Traumatol Turc,2014,48(2):217-222.
[2]Lee YJ.Vascular endothelial growth factor polymorphisms in patients with steroid-induced femoral head osteonecrosis[J].J Orthop Res,2012,30(1):21-27.
[3]Tong PJ.Research on the role of metabolism of fatty substance and osteoclast activity during the development of steroid-induced necrosis of femoral head[J].Zhongguo Gu Shang,2009,22(2):110-113.
[4]Jia YB.Inhibitory effects of vitamin E on osteocyte apoptosis and DNA oxidative damage in bone marrow hemopoietic cells at early stage of steroid-induced femoral head necrosis[J].Mol Med Rep,2017,15(4):1585-1592.
[5]Erken HY.Effect of pentoxifylline on histopathological changes in steroid-induced osteonecrosis of femoral head:experimental study in chicken[J].Int Orthop,2012,36(7):1523-1528.
[6]Lykissas MG.The role of hypercoagulability in the development of osteonecrosis of the femoral head[J].Orthop Rev(Pavia),2012,4(2):e17.
[7]Li GY.Edaravone,a novel free radical scavenger,prevents steroidinduced osteonecrosis in rabbits[J].Rheumatology(Oxford),2013,52(3):438-447.
[8]Zhang YL.Vitamin K2 Prevents Glucocorticoid-induced Osteonecrosis of the Femoral Head in Rats[J].Int J Biol Sci,2016,12(4):347-358.
[9]Liu X.A novel and facile synthesis of porous Si O2-coated ultrasmall Se particles as a drug delivery nanoplatform for efficient synergistic treatment of cancer cells[J].Nanoscale,2016,8(16):8536-8541.
[10]Koo KH.Risk period for developing osteonecrosis of the femoral head in patients on steroid treatment[J].Clin Rheumatol,2002,21(4):299-303.
[11]Song Q.Beneficial effect of grape seed proanthocyanidin extract in rabbits with steroid-induced osteonecrosis via protecting against oxidative stress and apoptosis[J].J Orthop Sci,2015,20(1):196-204.
[12]Lu PZ,Lai CY,Chan WH.Caffeine induces cell death via activation of apoptotic signal and inactivation of survival signal in human osteoblasts[J].Int J Mol Sci,2008,9(5):698-718.
[13]Chiappini F.Reactive oxygen species and extracellular signal-regulated kinase 1/2 mediate hexachlorobenzene-induced cell death in FRTL-5 rat thyroid cells[J].Toxicol Sci,2013,134(2):276-290.
[14]Choi EM,Kim GH,Lee YS.Diazoxide protects against hydrogen peroxide-induced toxicity in the osteoblastic MC3T3-E1 cells[J].Eur J Pharmacol,2009,624(1-3):45-50.
[15]Zhang JK.Protective effect of tetrahydroxystilbene glucoside against hydrogen peroxide-induced dysfunction and oxidative stress in osteoblastic MC3T3-E1 cells[J].Eur J Pharmacol,2012,689(1-3):31-37.
[16]Yang L.Stem cell factor(SCF)protects osteoblasts from oxidative stress through activating c-Kit-Akt signaling[J].Biochem Biophys Res Commun,2014,455(3-4):256-261.
[17]Huang SL,Jiao J,Yan HW.Hydrogen-rich saline attenuates steroid-associated femoral head necrosis through inhibition of oxidative stress in a rabbit model[J].Exp Ther Med,2016,11(1):177-182.
[18]Komurcu E.Preventive effects of coenzyme Q10(Co Q10)on steroid-induced osteonecrosis in rats[J].Acta Orthop Traumatol Turc,2014,48(2):217-222.