摘要
背景:激素性股骨头坏死与成骨细胞铁死亡密切相关,鹿角多肽可以通过抑制成骨细胞铁死亡,促进成骨细胞的存活与功能的建立,具有治疗激素性股骨头坏死的潜力,但其对成骨细胞铁死亡的调控机制尚未明确。目的:探究鹿角多肽抑制地塞米松诱导成骨细胞铁死亡的作用机制。方法:①采用不同浓度梯度的鹿角多肽与地塞米松干预MC3T3-E114细胞,CCK-8法检测细胞活性,确定鹿角多肽与地塞米松的作用浓度;②采用不同质量浓度梯度鹿角多肽干预被地塞米松(800μmol/L)处理后的MC3T3-E114细胞,分为空白对照组、地塞米松组、地塞米松+鹿角多肽组,采用CCK-8法计算不同质量浓度鹿角多肽对地塞米松(800μmol/L)干预MC3T3-E114细胞增殖的影响;③借助试剂盒检测空白对照组、地塞米松组、地塞米松+鹿角多肽组中超氧化物歧化酶活性及谷胱甘肽、丙二醛、脂质过氧化物、细胞铁、活性氧含量的变化,并应用Western blot检测谷胱甘肽过氧化物酶4、溶质载体家族7成员11蛋白的表达,验证鹿角多肽抑制铁死亡的通路。结果与结论:①CCK-8法检测细胞活性,结果确定后续实验选择以鹿角多肽(10 mg/mL)和地塞米松(800μmol/L)干预MC3T3-E114细胞处理24 h;②地塞米松干预后,丙二醛、脂质过氧化物、细胞铁、活性氧含量均升高(P<0.01),谷胱甘肽含量、超氧化物歧化酶活性均降低(P<0.01),谷胱甘肽过氧化物酶4、溶质载体家族7成员11蛋白表达降低(P<0.05-0.01);鹿角多肽干预后,上述各指标变化明显被逆转(P<0.05-0.01);③结果表明,鹿角多肽可能通过调控溶质载体家族7成员11/谷胱甘肽过氧化物酶4轴抑制成骨细胞铁死亡,发挥对激素性股骨头坏死的治疗作用。
BACKGROUND:Steroid-induced osteonecrosis of the femoral head is closely related to ferroptosis in osteoblasts.Deer antler peptide can promote the survival and functional establishment of osteoclasts by inhibiting ferroptosis in osteoblasts,and has the potential to treat steroid-induced osteonecrosis of the femoral head,but its regulatory mechanism of ferroptosis in osteoblasts has not yet been clarified.OBJECTIVE:To investigate the mechanism by which deer antler peptide inhibits dexamethasone-induced ferroptosis in osteoblasts.METHODS:(1)Different concentration gradients of antler peptide and dexamethasone were used to intervene in MC3T3-E114 cells,and the cell activity was detected by cell counting kit-8 method to determine the effect concentration of antler peptide and dexamethasone.(2)MC3T3-E114 cells treated with dexamethasone(800μmol/L)were intervened with different concentrations of gradient antler polypeptide,which were then divided into blank control group,dexamethasone group and dexamethasone+antler peptide group.Cell counting kit-8 method was used to calculate the effects of different concentrations of antler polypeptide on the proliferation of MC3T3-E114 cells.(3)Glutathione,superoxide dismutase,malondialdehyde,lipid peroxide,cellular iron,and reactive oxygen species levels in the blank control group,dexamethasone group and dexamethasone+antler peptide group were detected using kits.The protein expressions of glutathione peroxidase 4 and solute carrier family 7 member 11 were detected by western blot to verify the pathway by which antler polypeptide inhibits ferroptosis.RESULTS AND CONCIUSION:After cell activity was detected by cell counting kit-8 assay,antler peptide(10 mg/mL)and dexamethasone(800μmol/L)were selected to treat MC3T3-E114 cells for 24 hours in subsequent experiments.After treatment with dexamethasone,malondialdehyde,lipid peroxide,cellular iron and reactive oxygen species levels were all increased(P<0.01),while glutathione content and superoxide dismutase activity were decreased and the protein expression of glutathione peroxidase 4 and solute carrier family 7 member 11 were also decreased(P<0.05-0.01).After antler peptide intervention,the changes in the above indexes were obviously reversed(P<0.05-0.01).To conclude,antler peptide may inhibit ferroptosis in osteoblasts by regulating the glutathione peroxidase 4/solute carrier family 7 member 11 axis,and thereby exert a therapeutic role in steroid-induced osteonecrosis of the femoral head.
作者
邵学坤
王成
王仪
王平
邱卓雅
王欣如
孙铁锋
Shao Xuekun;Wang Cheng;Wang Yi;Wang Ping;Qiu Zhuoya;Wang Xinru;Sun Tiefeng(Shandong University of Traditional Chinese Medicine,Jinan 250355,Shandong Province,China;Shandong Academy of Chinese Medicine,Jinan 250014,Shandong Province,China;Qilu Institute of Technology,Jinan 273100,Shandong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2025年第14期2875-2881,共7页
Chinese Journal of Tissue Engineering Research
基金
山东省重点研发计划(科技示范工程)(2021SFGC1205),项目负责人:王平
济南市“新高校20条”资助项目引进创新团队计划(202333006),项目参与人:王平、王仪
济南市“新高校20条”资助项目(202228121),项目负责人:王平
泰山学者工程资助项目(tstp 20231239),项目参与人:王平。
关键词
鹿角多肽
地塞米松
激素性股骨头坏死
抑制
成骨细胞
铁死亡
antler peptide
dexamethasone
steroid-induced osteonecrosis of the femoral head
inhibition
osteoblast
ferroptosis
