摘要
目的探究微小核糖核酸-34a(microRNA-34A,miR-34a)基于蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路对骨关节炎大鼠的影响及作用机制。方法研究时间为2020年2月至2021年5月,选取43只6~9周龄健康雄性SD大鼠作前瞻性研究,其中随机选取10只分为空白组,剩余33只建立骨关节炎模型。成功建模30只,随机分为模型组、上调miR-34a组、沉默miR-34a组,各10只。观察病理组织,实时荧光定量PCR(real-time PCR,RT-PCR)法检测miR-34a表达,酶联免疫吸附实验法检测白细胞介素(interleukin,IL)-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IL-4、IL-10,蛋白免疫印迹法检测磷酸化蛋白激酶B(phosphorylated protein kinase B,pAKT)、磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin,pmTOR)表达。其中多组间比较行F检验,两组间比较行独立样本t检验。结果空白组、模型组、上调miR-34a组、沉默miR-34a组的miR-34a表达分别为(0.94±0.08)、(2.39±0.26)、(2.12±0.23)、(1.54±0.17),Makin评分分别为(0.35±0.05)、(4.13±0.19)、(3.02±0.16)、(1.15±0.09)分,IL-1β分别为(3.68±0.37)、(7.51±1.23)、(5.72±0.95)、(4.63±0.72)ng/L,TNF-α分别为(37.45±4.85)、(62.05±7.49)、(56.26±6.25)、(43.85±5.10)ng/L,IL-4分别为(115.75±16.81)、(50.43±9.42)、(75.82±11.94)、(93.85±13.86)pg/ml,IL-10分别为(106.49±17.51)、(60.85±10.23)、(76.81±13.07)、(91.05±15.02)pg/ml,pAKT分别为(1.00±0.01)、(1.95±0.29)、(1.71±0.26)、(1.71±0.26),pmTOR分别为(1.00±0.01)、(1.89±0.24)、(1.56±0.25)、(1.25±0.15)。与空白组相比,模型组、上调miR-34a组、沉默miR-34a组的miR-34a表达、Makin评分、IL-1β、TNF-α、pAKT、pmTOR较高,IL-4、IL-10较低,差异均有统计学意义(F/P值分别为25.290/0.001、91.260/0.001、14.144/0.001、13.077/0.001、15.525/0.001、17.580/0.001、16.080/0.001、10.676/0.001);与模型组相比,上调miR-34a组、沉默miR-34a组miR-34a表达、Makin评分、IL-1β�
Objective To explore the effect of microrNA-34A(miR-34a)on osteoarthritis in rats based on the AKT/mTOR pathway and its mechanism.Methods The study was form February 2020 to May 2021.In this study,43 healthy male SD rats were selected for prospective study,and they were 6-9 weeks old.Among them,10 rats were randomly selected as a blank group,and The remaining 33 rats were established as the osteoarthritis models.The 30 rats successfully modeled were randomly divided into a model group,an up-regulated miR-34a group,and a silenced miR-34a group,with 10 rats in each group.The expression of miR-34a was detected by real-time polymerase chain reaction(RT-PCR),and The levels of interleukin-1β(Il-1β),tumor necrosis factor-α,TNF-α,interleukin-4(IL-4),and interleukin-10(IL-10)was detected by enzyme-linked immunosorbent assay(ELISA).The expressions of phosphorylated protein kinase B(pAKT)and phosphorylated mammalian target of rapamycin(pmTOR)were detected by Western blot.F test was used for the comparison between≥3 groups,and independent-sample t test was used for the comparison between two groups.Results The expressions of miR-34a in the blank group,the model group,the up-regulated miR-34a group,and the silenced miR-34a group were(0.94±0.08),(2.39±0.26),(2.12±0.23),and(1.54±0.17),rthe Makin scores(0.35±0.05),(4.13±0.19),(3.02±0.16),and(1.15±0.09),the IL-1βlevels(3.68±0.37),(7.51±1.23),(5.72±0.95),and(4.63±0.72)ng/L,the TNF-αlevels(37.45±4.85),(62.05±7.49),(56.26±6.25),and(43.85±5.10)ng/L,the IL-4 levels(115.75±16.81),(50.43±9.42),(75.82±11.94),and(93.85±13.86)pg/ml,the IL-10 levels(106.49±17.51),(60.85±10.23),(76.81±13.07),and(91.05±15.02)pg/ml,the pAKT(1.00±0.01),(1.95±0.29),(1.71±0.26),and(1.71±0.26),and the pmTOR(1.00±0.01),(1.89±0.24),(1.56±0.25),and(1.25±0.15).Compared with those in the blank group,the expression of miR-34a,Makin score,IL-1β,TNF-α,pAKT and pmTOR were higher in than those in the model group,the up-regulated miR-34a group,and the silenced miR-34a group,while IL-
作者
周伦
丁源
吕洲
Zhou Lun;Ding Yuan;Lyu Zhou(Department of Osteoarthrology,Qingdao Municipal Hospital,Qingdao 266000,China)
出处
《国际医药卫生导报》
2022年第21期2990-2994,共5页
International Medicine and Health Guidance News
