目的 探讨微小RNA-381-3p(MicroRNA-381-3p,miR-381-3p)对脊髓损伤(Spinal cord injury,SCI)大鼠继发性损伤的修复机制以及其对肿瘤抑制蛋白基因(Tumor protein p53,TP53)表达水平的调控机制.方法 双荧光素酶以及蛋白质免疫共沉淀检测miR-381-3p和TP53的作用;用改良Allen's重物垂直撞击法复制大鼠SCI模型;实验动物分为假手术(Sham)组、SCI组、miR-381-3p-mimic(mimic)组、mimic+pc DNA3.1 TP53(mimic+pc)组,每组各 10 只;mimic 组大鼠尾静脉注射 miR-318-3p mimic;mimic+pc 组大鼠尾静脉注射 miR-318-3p mimic 和 pc DNA3.1 TP53;Sham 组和 SCI 组大鼠尾静脉注射 miR-318-3p mimic-NC和pc DNA3.1 TP53-NC;检测各组大鼠的运动功能评分(Basso beattie bresnahan,BBB)评分;脱氧核糖核苷酸末端转移酶介导的缺口末端标记试剂盒(Terminal-deoxynucleoitidyl transferase-mediated nick end labeling,TUNEL)检测各组大鼠脊髓组织的细胞凋亡;超氧化物阴离子二氢乙啶(Dihydroethidium,DHE)荧光探针检测脊髓组织中活性氧(Reactive oxygen species,ROS)自由基的水平;免疫荧光检测各组大鼠脊髓组织中小胶质细胞的表型转换;酶联免疫吸附(Enzyme-linked immunosorbent assay,ELISA)检测各组大鼠血清中超氧化物歧化酶(Superoxide dismutase,SOD)、丙二醛(Malondialdehyde,MDA)、白细胞介素-1β(Interleukin-1 β,IL-1β),IL-6,IL-4和IL-10的水平;彗星实验检测各组脊髓组织中的DNA损伤;Western blot实验检测各组大鼠脊髓组织中肿瘤抑制蛋白基因(Tumor protein p53,TP53)、核转录因子(Nuclear factor κB,NF-κB)、磷酸化组蛋白(γ-Histone family 2A variant,γ-H2AX)、B 淋巴细胞瘤-2(B cell lymphoma-2,Bcl-2)和 Bcl-2 相关 X 蛋白(Bcl2-associated × protein,Bax)水平.结果 双荧光素酶以及蛋白质免疫共沉淀显示miR-381-3p调控TP53的表达水平.与Sham组比较,SCI组、mimic组、mimic+pc组大鼠的BBB评分、血清中SOD的活性、脊髓组织中M2型细胞的比例、Bcl-2的表达水平明显降低,脊髓组织中的细胞凋亡率、ROS的水平,血清中MDA,IL-1β,IL-6,IL-4和IL-1 0的水平,脊髓组织中M1型细胞的比例、DNA的Olive尾矩、TP53,NF-κB,γH2AX和Bax的表达水平明显升高(P均＜0.05);与SCI组比较,mimic组、mimic+pc组大鼠的BBB评分、血清中SOD的活性以及IL-4和IL-10的水平、脊髓组织中M2型细胞的比例、Bcl-2的表达水平明显升高,脊髓组织中的细胞凋亡率、ROS的水平,血清中MDA,IL-1β,IL-6的水平,脊髓组织中M,型细胞的比例、DNA的Olive尾矩、TP53,NF-κB,yH2AX和Bax的表达水平明显降低(P均＜0.05);与mimic组比较,mimic+pc组大鼠的BBB评分、血清中SOD的活性以及IL-4和IL-10的水平、脊髓组织中M2型细胞的比例、Bcl-2的表达水平明显降低,脊髓组织中的细胞凋亡率、ROS的水平、血清中MDA,IL-1β,IL-6的水平,脊髓组织中M1型细胞的比例、DNA的Olive尾矩、TP53,NF-κB,γH2AX和Bax的表达水平明显升高(P均＜0.05).结论 SCI大鼠模型miR-318-3p的表达水平明显降低,TP53的表达水平明显升高,miR-318-3p靶向TP53的表达水平,过表达miR-318-3p后能明显抑制实验动物脊髓组织中的氧化和炎症应激,减轻细胞的DNA损伤,降低脊髓组织中的细胞凋亡率,缓解动物脊髓组织的病理损伤.

Spinal cord injury (SCI) is a severe neurological disorder and the molecular mechanisms leading to its poor prognosis remain to be elucidated. S100A1, a mediator of Ca2+ handling of sarcoplasmic reticulum and mitochondrial function, operates as an endogenous danger signal (alarmin) associated with inflammatory response and tissue injury. The aim of the present study was to investigate the expression and biological effects of S100A1 in SCI. A rat model of SCI and a PC12 cell model of lipopolysaccharide (LPS)-induced inflammation were established to examine S100A1 expression at the mRNA and protein levels. The inflammation level, which was mediated by S100A1, was determined based on inflammatory factor (IL-1 beta, IL-6 and TNF-alpha) and anti-inflammatory factor (IL-10) expression. The effects of S100A1 on cellular oxidation and anti-oxidation levels were observed by detecting the levels of reactive oxygen species, superoxide dismutase, catalase activities and nuclear factor erythroid 2-related factor 2 expression. The protein levels of Bax, Bcl2 and cleaved caspase-3 were used for the evaluation of the effects of S100A1 on apoptosis. Phosphorylated (p-)ERK1/2 expression was used to evaluate the effects of S100A1 on ERK signaling. The results revealed that S100A1 expression was significantly upregulated in vivo and in vitro in the PC12 cell model of LPS-inflammation. The silencing and overexpression of S100A1 helped alleviate and aggravate LPS-induced inflammation, oxidative stress and apoptosis levels, respectively. S100A1 was found to regulate the ERK signaling pathway positively. An inhibitor of ERK signaling (MK-8353) partially abolished the promoting effects of the overexpression of S100A1 on inflammation, oxidative stress damage and apoptosis. In conclusion, S100A1 expression was elevated in model of SCI and in the PC12 cell model of LPS-induced inflammation. Furthermore, the overexpression/silencing S100A1 aggravated/mitigated the inflammation, oxidative stress damage and the apoptosis of LPS-stimulated PC12 cells via the ERK signaling pathway. The present study revealed the mechanism of S100A1 in SCI, which provided a new theoretic reference for future research on SCI.

Nowadays, there is a serious lack of information about the value-added apoptosis of sarcoma cells in China. Especially in clinical medicine, exploring the effect of ibuprofen on the growth and apoptosis of fibrosarcoma cells under the PI3K/Akt/mTOR signaling pathway can not only effectively prevent us in advance, but also be a great way to break through this field. The main purpose of this study was to investigate the effects of ibuprofen on the proliferation, cell cycle and apoptosis of fibrosarcoma cells through the PI3K/Akt/mTOR signaling pathway. We divided the HTl080 cell line into zero control group, control group and experimental group. The withering group was not inoculated with any cells, while the control group was only added with the same amount of culture medium, while the experimental group was added with 5,10,15,20 concentrations respectively. We found that the apoptosis rate of sarcoma cells in the control group increased from 5.66% to 7.12%, while the apoptosis rate of sarcoma cells in the experimental group increased significantly faster than that in the control group, with an overall increase of 7.16%, from 4.56% to 11.72%. Therefore, we can be surer that ibuprofen has a very good inhibitory effect on the proliferation, cell cycle and apoptosis of fibrosarcoma cells under the PI3K/Akt/mTOR signaling pathway. Therefore, when ibuprofen was injected into the body, it could not only observe the sarcoma cells well but also reflect the good inhibitory effect of ibuprofen on other substances in vivo under the PI3K/Akt/mTOR signaling pathway.

Osteosarcoma is an aggressive cancer of the skeletal system, which is associated with a poor prognosis due to the high recurrence rate. Although previous studies have revealed that competitive endogenous RNAs (ceRNAs) are involved in various biological processes in the physiology and development of osteosarcoma, the roles of ceRNAs in osteosarcoma recurrence remain largely unexplored. The present study constructed a ceRNA-ceRNA network for osteosarcoma by systematically integrating matched expression profiles for microRNAs (miRNAs/miRs) and mRNAs, and identified two ceRNA-mediated modules that were associated with recurrence in patients with osteosarcoma. A multivariate Cox regression analysis demonstrated that the recurrence-free prognosis associated with the expression of the two modules was independent of other clinical factors. In addition, hsa-miR-335-3p was identified as an upstream regulating factor for both modules. In conclusion, the results of the present study suggested that ceRNAs may act as potential therapeutic biomarkers for predicting the recurrence of osteosarcoma, and may help to identify patients with osteosarcoma at a high risk of recurrence, who may benefit from adjuvant therapy.