摘要
目的 探讨miR-125a-3p对骨关节炎大鼠软骨细胞活性、氧化应激反应及细胞沉默信息调节因子1/叉头框转录因子o1(SIRT1/Foxo1)蛋白表达的影响。方法 将40只大鼠随机分为对照组(健康大鼠)、模型组(骨关节炎大鼠,造模后不做其他处理及干预)、miR-125a-3p组(骨关节炎大鼠,造模成功7 d后于大鼠左膝关节关节腔内注射miR-125a-3p抑制剂)、阿利吉仑组(骨关节炎大鼠,造模成功7 d后给予大鼠50 mg/kg阿利吉仑灌胃),每组10只。通过氧化应激反应检测超氧物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、乳酸脱氢酶(LDH),HE、番红O、Masson染色检查大鼠软骨损伤情况,TUNEL检测软骨细胞凋亡,RT-PCR与免疫印迹检测SIRT1、Foxo1基因及蛋白水平。结果 软骨细胞凋亡率模型组高于miR-125a-3p组与阿利吉仑组(P<0.05),miR-125a-3p组与阿利吉仑组比较差异无统计学意义(P>0.05)。HE、番红O、Masson染色显示,模型组大鼠关节软骨面骨裂隙形成;miR-125a-3p组与阿利吉仑组大鼠关节软骨表面变得光滑,无明显裂隙。RT-PCR与免疫印迹检测显示,与模型组相比,miR-125a-3p组与阿利吉仑组SOD、GSH、SIRT1水平均升高(P<0.05), MDA、LDH、Foxo1水平均降低(P<0.05)。结论 miR-125a-3p通过调控SIRT1/Foxo1信号通路,可降低氧化应激反应,抑制软骨细胞凋亡,改善骨关节炎病情。
Objective To investigate the infulence of miR-125a-3p on the activity,oxidative stress reaction and silent information regulator 1/forkhead transcription factor o1(SIRT1/Foxo1)protein of chondrocytes in osteoarthritic rats.Methods The 40 rats were randomly divided into control group(healthy rats),model group(osteoarthritis rats were not given other treatment and intervention after successful modeling),miR-125a-3p group(osteoarthritis rats,at 7 d after the success of the modeling,the left knee joint cavity of the rats were injected with miR-125a-3p inhibitor),Aliskiren group(osteoarthritis rats,the rats were given 50 mg/kg Alegiren by gavage at 7 d after successful modeling),with 10 rats in each group.Superoxide dismutase(SOD),malondialdehyde(MDA),glutathione(GSH)and lactate dehydrogenase(LDH)were detected by oxidative stress reaction.HE,saffron O and Masson staining were used to examine the cartilage injury of rats.Chondrocyte apoptosis was detected by TUNEL.The gene and protein levels of SIRT1 and Foxo1 were detected by RT-PCR and Western Blotting method.Results The apoptosis rate of chondrocytes:the model group was higher than that in the miR-125a-3p group and the Aliskiren group(P<0.05),but there was no significant difference between the miR-125a-3p group and the Aliskiren group(P>0.05).HE,saffron O and Masson staining showed:for the surface of articular cartilage of the rats,there were bone fracture formation in model group;and which became smooth in miR-125a-3p group and Aliskiren group,without obvious fracture.RT-PCR and Western Blotting test showed:compared with the model group,the levels of SOD,GSH,SIRT1 in miR-125a-3p group and Aliskiren group were increased(P<0.05),while the levels of MDA,LDH,Foxo1 were decreased(P<0.05).Conclusions miR-125a-3p can reduce oxidative stress reaction,inhibit chondrocyte apoptosis and improve osteoarthritis situation by regulating SIRT1/Foxo1 signaling pathway.
作者
王乐
WANG Le(Dept of Orthopaedics,the People′s Hospital of Haikou City,Haikou Hospital Affiliated to Xiangya Medical College of the Central South University,Haikou,Hainan 570000,China)
出处
《临床骨科杂志》
2023年第2期293-298,共6页
Journal of Clinical Orthopaedics
