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Objective:To study the changes of plasma receptor interacting protein 3 (RIP3) levels in neonatal late-onset sepsis (LOS) and to determine its clinical value.Methods:From October 2019 to April 2021, plasma samples and clinical data of LOS infants admitted to our hospital were prospectively studied. Infants with similar gestational ages admitted for non-infectious diseases were assigned into the control group. Enzyme-linked immunoassay was used to determine plasma RIP3 levels. The clinical value of plasma RIP3 in the diagnosis and treatment of neonatal LOS were analyzed.Results:A total of 152 cases (76 in the LOS group and 76 in the control group) were included in the study. No significant differences existed in the baseline data between the two groups. A total of 226 plasma samples were collected (76 samples from the LOS group before treatment, 74 samples after treatment and 76 samples from the control group). The plasma RIP3 level of LOS group before treatment (19.9±6.3 ng/ml) was significantly higher than the control group (11.4±3.5 ng/ml) and the after treatment group (11.9±3.5 ng/ml) ( P<0.05). The plasma RIP3 level had good diagnostic value for neonatal LOS (AUC=0.884). With cut-off value of 15.5 ng/ml, the plasma RIP3 showed the best diagnostic efficacy (Youden index 0.658, sensitivity 72.4%, specificity 93.4%, positive likelihood ratio 11.0, negative likelihood ratio 0.3). Conclusions:Plasma RIP3 level is closely related with neonatal LOS and may be used for the early diagnosis and therapeutic evaluation of neonatal LOS.
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Myocardial ischemia/reperfusion (I/R) injury seriously endangers human health and is a potential hidden danger in the treatment of cardiovascular diseases, among which myocardial necrosis is one of the mechanisms of myocardial I/R injury. Numerous studies have shown that necrosis factor is widely involved in the regulation of myocardial cell necrosis, but its specific mechanism is not fully understood. Receptor interacting protein 3/receptor interacting protein kinase 3 (RIP3/RIPK3) is an essential protein in necroptosis pathways, and activated RIP3 can cause irreversible necrosis of myocardial cells. On the basis of introducing RIP3 molecule, this paper focused on the role and mechanism of RIP3 mediated necroptosis pathway in myocardial cell necroptosis, with a view to providenew ideas and insights for the treatment of myocardial I/R injury.
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AIM: To explore whether tumor necrosis factor-α (TNF-α) induces necroptosis in murine long bone osteocyte-like cell line MLO-Y4 and the possible mechanism.METHODS: The MLO-Y4 cells were divided into control group, TNF-α group, TNF-α+necrostatin-1 (Nec-1) group, TNF-α+Z-VAD group and TNF-α+receptor-interacting protein 3 (RIP3)-siRNA group.The death rate of MLO-Y4 cells was assessed by flow cytometry with Annexin V-FITC/PI staining.The morphological features of the cells were observed under transmission electron microscope (TEM).The protein levels of RIP1, RIP3 and cleaved caspase-3 were determined by Western blot.Finally, the numbers of total cells and RIP1-RIP3-positive cells were observed under laser scanning confocal microscope.The production of reactive oxygen species (ROS) in the cells was measured by DCFH-DA staining.RESULTS: Compared with control group, the apoptotic or necroptotic rate of the cells induced by TNF-α was increased significantly (P<0.01).The increased apoptotic or necroptotic rate was dramatically reduced by treating with Nec-1, Z-VAD or RIP3-siRNA transfection (P<0.01).In TNF-α group and TNF-α+Z-VAD group, a lot of MLO-Y4 cells with typical necroptotic morphological features were observed under TEM.However, obvious necroptotic cells were not found in Nec-1 or RIP3-siRNA treatment group.The protein level of RIP1 in the cells treated with Nec-1 was sharply lower than that in TNF-α group (P<0.01).However, Z-VAD did not reduce the elevated levels of RIP1 and RIP3.RIP3-siRNA effectively down-regulated the protein level of RIP3 compared with TNF-α group (P<0.01).Nec-1 effectively down-regulated the protein levels of RIP1 colocalized with RIP3 compared with TNF-α group (P<0.01).However, Z-VAD did not reduce the levels of RIP1 colocalized with RIP3.Nec-1, Z-VAD and RIP3 siRNA significantly decreased the ROS levels (P<0.01).CONCLUSION: TNF-α induces the necroptosis of MLO-Y4 cells.RIP3 play vital roles in the cell necroptotic signal pathway.ROS may be the executor of necroptosis of MLO-Y4 cells.
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Objective To investigate the location of receptor interacting protein 3( RIP3) in Necroptosis and its function in this signal passage, and explore the relationship between receptor interacting protein 1 ( RIP1 ) and RIP3 in nuclear translocation. Methods Primary cerebrocortical neurons were cultured for 12 days,then pre-treated with zVAD-fmk(20μ,mol/L) for half an hour to block apoptosis. ①Extracting nuclear and cytoplasmic protein after neurons were exposed to TNF for different time ,then protein levels of RIP3 were analyzed by western blot and immunofluorescence for qualitative observation;②In the following research,the neurons were treated with Nec-1 and shRlPl ,then the protein level of RIP1 and RIP3 with western blot were analyzed, cell viability were determined by measuring LDH levels. Results ①In signaling pathways of necroptosis, the protein level of RIP3 in cytoplasmic decreased gradually with prolonged TNF exposure, to the corresponding it rolled up in nucleus and a-chieved the peak in 12 hours of TNF treatment ( Cytoplasmic 0. 45 ± 0. 03 ,0. 41 ± 0. 02,0. 73 ± 0. 03 ,0. 90 ± 0.01,1.15 ±0.04,1.30 ±0.02,0.99 ±0.03,0.63 ±0. 03;Nucleus 0. 07 ±0.02,0. 26 ±0.02,0. 57 ±0. 02,0. 68 ± 0.02,0. 80 ± 0.01,0.92 ± 0.02,1.28 ± 0.03,0. 87 ± 0.02) (P < 0.01). ②Blocking the relationship between RIP1 and RIP3 with necrostatin-1 and shRIPl , nuclear translocation of RIP3 decreased and caused a great increase in cell viability( 1.00 ±0.05,0.39 ±0.03,0.50 ±0. 03) (P<0. 01). Conclusion RIP3 mainly locates in cy-tolymph of normal cells,it translocates into nucelus as necroptosis takes place. RIP1 function with RIP3 in nuclear translocation. Block nuclear translocation of RIP3 is a potential way to protect cells.
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Objective To study the molecular mechanism of the remission in children with acute lymphocyte leukemia(ALL),as well as the expression of thyroid hormone receptor interacting protein 3(TRIP3) gene in children with ALL and explore the relationship between TRIP3 and ALL.Methods Fasting venous blood 2-4 mL was collected,anticoagulanted with ethylenediamine tetraacetic acid(EDTA),then perpheral blood mononuclear cells was collected by Ficoll density gradient centrifugation,total RNA was extracted by Trizol one step method.Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect TRIP3 expression in peripheral blood lymphocytes in 73 ALL children of different stages and 20 normal children.The relationship between TRIR3 expression in children and ALL release was analyzed.Results 1.Expression of TRIP3 was significantly lower both after initial treatment and during recrudescence than that in normal children(Pa0.05).3.In children with ALL after initial treatment,the remission rate was significantly higher in TRIP3 positive patients than in TRIP3 negative ones(remission rate discern 25.0% vs 84.2%,P