Introduction: This study aimed to elucidate the neuroprotective mechanism of Isorhynchophylline (IRN) in stroke. Methods: Using network pharmacology, the action targets of IRN were predicted using the SwissTargetPrediction, HERB, and TCMIP 2.0 databases. We identified sixty-five targets common to IRN and stroke. We conducted GO and KEGG enrichment analyses using R software and constructed a compound- target- pathway- disease network. Additionally, molecular docking experiments were performed to investigate the binding interactions between IRN and its main targets. Subsequently, an oxygen-glucose deprivation/reoxygenation (OGD/R) model was established using the human neuroblastoma cell line SH-SY5Y under hypoxic conditions (1% O2). The effective working concentration of IRN was determined by measuring cell viability using the CCK-8 assay. Then, an ELISA experiment was conducted to detect the expression of inflammatory factors IL-1β and IL-10. The activity of reactive oxygen species (ROS) was measured using the dual fluorescence detection method. TUNEL staining was employed to assess the apoptosis of cells. Finally, a Western blot was used to examine the effect of IRN on EGFR and AKT expression in cells. Results: The results of network pharmacology analysis indicated that the main targets of IRN in treating stroke are EGFR, mTOR, JAK2, AKT, and JUN. Enrichment analysis revealed that IRN is associated with neurotransmitter receptor function and participates in the EGFR signaling pathway. Molecular docking results showed that it has a particularly strong binding affinity for AKT1 (binding energy ≈ -8.7 kcal/mol). The in vitro experiments showed that IRN treatment significantly alleviated the damage induced by OGD/R. Specifically, the levels of IL-1β and ROS decreased, the level of IL-10 increased, and the apoptosis rate decreased. Western blot results showed that, compared with the OGD/R group, IRN significantly upregulated the expression of p-EGFR and p-AKT. We demonstrated that IRN employs a multi-target approach to counteract stroke pathology, as revealed by computational predictions and experimental validation. Discussion: The convergence of network pharmacology, molecular docking, and in vitro assays enabled the identification of key targets, including EGFR, AKT1, and JAK2. These methods also indicate that IRN can simultaneously regulate the inflammatory, oxidative stress, and apoptotic processes. Compared to singletarget agents, IRN exhibits superior multi-target therapeutic potential. These findings, derived from database analyses and cell-based assays, provide a comprehensive overview of IRN's pharmacological effects, which are consistent with its known neuroprotective benefits. Although these results are obtained from cell-based models and predictive databases, they provide a cohesive mechanistic rationale that is consistent with the neuroprotective properties ascribed to IRN. Further studies in animal models and in clinical settings must be conducted to apply these insights for therapeutic purposes. Conclusion: IRN can exert neuroprotective effects against stroke by regulating key targets, including EGFR. It exhibits anti-inflammatory, anti-oxidative, and anti-apoptotic activities, which provide a reasonable basis for developing IRN as a drug for treating stroke.

BACKGROUND:Age, biomarkers including pepsinogen and gastrin, and Helicobacter pylori (H pylori) might be risk factors in nodular gastritis (NG), however, the detailed relationship between the risk factors and inflammation in NG remains explored. Thus, we aimed to determine the association of age, pepsinogen and gastrin levels and NG patients infected by H pylori. METHODS:A total of 768 cases of NG patients were identified in the endoscopic procedure records spanning from 2019 to 2023 at the Baotou Medical College, who were divided into 7 age-based groups: 8 to 17, 17 to 30, 31 to 40, 41 to 50, 51 to 60, 61 to 70, and 71 to 80 years. Results pertaining to serum gastric function test, endoscopic findings, histological examinations, and C13 urea breath tests were systematically compiled from the patients' medical records. RESULTS:The prevalence of 768 NG patients exhibited a "parabolic pattern" with increasing age. Peaks were observed 41 to 50 group, especially in female patients. Among these patients, 73.6% were female patients. Besides, there was a significant difference in pepsinogen I (PGI)/pepsinogen II (PGII) ratio between the mild infection and the others group. Additionally, a similar significant difference was observed between the moderate infection group and the other groups. The logistic multiple regression analysis facilitated the combined assessment of various biomarkers such as PGI, PGII, PGI/PGII, and gastrin-17 (G-17), resulting in a sensitivity of 73.7% and a specificity of 97.9%. CONCLUSIONS:NG patients demonstrated a parabolic distribution pattern based on age. And, the elevated levels of PGI, PGII and G-17 observed in patients could signify their contributory role in the initiation and progression of NG.

Prior research has firmly established that the N-methyl-d-aspartate (NMDA) receptor subunit 2 B (GluN2B) and its phosphorylation contribute to ischemic/hypoxic brain injury. Hypoxic preconditioning (HPC) is an endogenous mechanism that protects the brain from both ischaemic and hypoxic damage. In this study, we explored the effects of HPC on GluN2B and its phosphorylation at two sites (tyrosine residues 1252 and 1336), catalysed by Fyn, in the hippocampus both in vivo and in vitro. Animal and cellular models of HPC were developed by subjecting mice and the mouse hippocampal neuronal cell line HT22 to repeated hypoxia. Levels of GluN2B and its phosphorylation at the tyrosine residues 1336 (pY1336 GluN2B) and 1252 (pY1252 GluN2B) were detected in HPC-treated hippocampi and HT22 cells using western blotting and immunofluorescence. The distributions of GluN2B, pY1336 GluN2B, and pY1252 GluN2B in the synaptic (TxP) and extrasynaptic components (TxS) were analysed by western blotting. Caspase-3 and spectrin, both markers of cellular injury, were further measured using western blotting. HPC downregulated GluN2B and pY1336 GluN2B levels in the hippocampus and HT22 cells. The changes in GluN2B and pY1336 GluN2B levels in the extrasynaptic components were similar to those in the hippocampus and HT22 cells, while the changes in the synaptic components showed the opposite trend which increased after HPC. The downregulation of GluN2B and pY1336 GluN2B may be associated with neuroprotection induced by HPC. Additionally, their localization at synaptic and extrasynaptic sites may play distinct roles in neuroprotection.

Myocardial infarction (MI) is a leading cause of death and disability worldwide. The promotion of myocardial regeneration is a promising therapeutic strategy for acute MI. Using a zebrafish ventricular ablation system, we found that the Mongolian traditional medicine Eerdun-Wurile (EW) promotes myocardial regeneration in zebrafish. EW treatment significantly accelerated proliferation of myocardial cells and improved cardiac function. Transcriptome sequencing revealed a significant decrease in mevalonate diphosphate decarboxylase a (mvda) expression in the metronidazole-induced ventricular ablation group, whereas mvda expression was restored in the EW group. mvda knockdown using morpholino oligonucleotides reversed the EW-mediated myocardial regeneration, whereas mvda overexpression enhanced the regenerative ability. In conclusion, EW may promote zebrafish myocardial regeneration, accelerate myocardial cell proliferation, and improve cardiac function by upregulating mvda expression. Our data partially revealed the molecular mechanism by which EW promotes myocardial regeneration and repair, and provides experimental data and novel insights for advancing MI treatment.

Depression, a neurological disorder triggered by stressful stimuli such as hypoxia, is associated with high morbidity and mortality. Hypoxic preconditioning (HPC) is an endogenous mechanism that has been used in recent research to upregulate BDNF, a marker of depression, to elicit neuroprotective effects. However, the mechanisms by which HPC protects against depression remain poorly understood. Therefore, this study aimed to investigate the effects of HPC on depressive behaviors via BDNF signaling. Initially, ICR mice were subjected to HPC, followed by the establishment of a 24-hour restraint stress model to mimic depressive behaviors. Subsequent analysis focused on changes in depressive behaviors, biochemical markers, and the levels of BDNF and its ability to modulate synaptic structure and neurogenesis. Furthermore, whole transcriptome sequencing was conducted. The results indicated that HPC relieved characteristic depressive behaviors in restraint stress model mice, regulated neurotransmitter levels, elevated antioxidant capacity, and promoted BDNF signaling in the hippocampus. PSD-95 expression, the number and complexity of neuronal dendritic spines, and hippocampal neurogenesis in model mice were increased via HPC. Restraint stress regulated 373 DElncRNAs, 166 DEcircRNAs, 29 DEmiRNAs and 1235 DEmRNAs, which were also modulated by HPC. The ceRNA networks were constructed on the basis of these DERNAs. Functional enrichment analysis revealed that these genes are related to synapses, neurogenesis and neurotrophin signaling. These results suggested that HPC upregulated BDNF and activated BDNF/PLCγ/CREB signaling to alleviate synaptic deficits and promote hippocampal neurogenesis, ultimately ameliorating depressive behaviors in mice. The identification of various mRNAs and ncRNAs and their constituent ceRNAs provides theoretical guidance for the clinical treatment of depression with HPC.

The N-methyl-D-aspartate (NMDA) receptors are related to the various functioning of the nervous system. It has been shown that the NR2B subunit plays an important role in neurological hypoxic/ischemic diseases by regulating NMDA receptor function. NR2B tyrosine phosphorylation is also an important regulatory mechanism for NMDA receptor function. However, the mechanism of NR2B tyrosine phosphorylation in hypoxic/ischemic injury is still unclear. Therefore, in the present study, we aimed to further clarify the changes in NR2B tyrosine phosphorylation in hypoxic/ischemic damage in the brain and its relationship with neuronal survival under hypoxic/ischemic conditions. Four types of NR2B tyrosine site mutants (Tyr → Phe at 1252, 1336, and 1472, and all three mutations together, named Y1252F, Y1336F, Y1472F, and Triple) and wild-type plasmids were transfected into HT22 cells. The cells were then exposed to oxygen–glucose deprivation and reoxygenation (OGD/R). NR2B, cell apoptosis-related molecules, and neuronal survival factor CREB-related signaling proteins (CaMKII, ERK, Akt) were measured. Cell viability was assessed using the CCK-8 assay. Cell apoptosis and cell cycle were evaluated using flow cytometry. The death ratio of HT22 cells under OGD conditions was further tested using a live cell analysis platform. The viability of HT22 cells in the Y1252F, Y1336F, Y1472F, Triple mutants, and wild-type groups was elevated. Compared to the wild-type, western blotting and real-time PCR showed that Y1252F, Y1336F, Y1472F, and Triple mutants downregulated the expression of apoptosis factors and upregulated anti-apoptosis factors in the OGD/R model. Flow cytometry and cell cycle analysis demonstrated that Y1252F, Y1336F, Y1472F, and Triple mutants reduced the apoptosis rate. The percentage of cells in the S phase decreased significantly. Live cell analysis illustrated that the Y1252F, Y1336F, Y1472F, and Triple mutants contributed to HT22 cell survival under OGD conditions. Additionally, the Y1252F, Y1336F, Y1472F, and Triple mutants activated the survival signaling pathway. Furthermore, compared to the control group (without plasmid), only the Y1336F, Y1472F, and Triple mutants groups showed significant differences in the above tests. The tyrosine phosphorylation of NR2B at Y1336 and Y1472 plays key roles in hypoxic/ischemic injury. These phosphorylation sites may be potential targets for hypoxic/ischemic neural protection.

Damage to the ribosome or an imbalance in protein biosynthesis can lead to some human diseases, such as diabetic retinopathy (DR) and other eye diseases. Here, we reported that the kri1l gene was responsible for retinal development. The kri1l gene encodes an essential component of the rRNA small subunit processome. The retinal structure was disrupted in kri1l mutants, which resulted in small eyes. The boundaries of each layer of cells in the retina were blurred, and each layer of cells was narrowed and decreased. The photoreceptor cells and Müller glia cells almost disappeared in kri1l mutants. The lack of photoreceptor cells caused a fear of light response. The development of the retina started without abnormalities, and the abnormalities began two days after fertilization. In the kri1l mutant, retinal cell differentiation was defective, resulting in the disappearance of cone cells and Müller cells. The proliferation of retinal cells was increased, while apoptosis was also enhanced in kri1l mutants. γ-H2AX upregulation indicated the accumulation of DNA damage, which resulted in cell cycle arrest and apoptosis. The kri1l mutation reduced the expression of some opsin genes and key retinal genes, which are also essential for retinal development.

目的:观察蒙药额尔敦-乌日勒对斑马鱼心脏损伤后再生作用及其可能作用机制,验证其治疗的有效性,为进一步研究蒙药额尔敦-乌日勒提供实验依据.方法:利用转基因鱼系Tg(vmhc:mCherry-NTR)经甲硝唑(MTZ)处理后建立心室损伤模型,把 90 只受精 3d(3dpf,days post fertilization)的斑马鱼胚胎随机分为对照组(DMSO,n =30)和心肌消融组(MTZ,n = 30)以及心室损伤后额尔敦-乌日勒治疗组(MTZ+EW,n =30),治疗组在MTZ处理后24 h(24 hpt,24 hour post treatment)用蒙药额尔敦-乌日勒混悬液以25 μg/mL治疗.3 组在72 hpt检测细胞增殖,用共聚焦显微镜观察细胞增殖情况并定量分析,同时应用RT-qPCR分析一氧化氮合酶(NOS)、诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1 β)、白细胞介素-6(IL-6)的表达量.结果:治疗组斑马鱼胚胎经过蒙药额尔敦-乌日勒治疗后,心肌损伤后的再生修复比例明显上升,且再生心肌细胞的增殖速度较对照组加快、炎症因子表达量明显低于对照组.结论:蒙药额尔敦-乌日勒能够促进斑马鱼胚胎心肌细胞的修复再生,并促进心肌细胞的增殖,抑制心室损伤后的炎症反应.

Hypoxic preconditioning (HPC) as an endogenous mechanism can resist hypoxia/ischemia injury and exhibit protective effects on neurological function including learning and memory. Although underlying molecular mechanisms remain unclear, HPC probably regulates the expression of protective molecules by modulating DNA methylation. Brain-derived neurotrophic factor (BDNF) activates its signaling upon binding to the tropomyosin-related kinase B (TrkB) receptor, which is involved in neuronal growth, differentiation, and synaptic plasticity. Therefore, this study focused on the mechanism by which HPC regulates BDNF and BDNF/TrkB signaling through DNA methylation to influence learning and memory. Initially, the HPC model was established by hypoxia stimulations on ICR mice. We found that HPC downregulated the expression of DNA methyltransferase (DNMT) 3A and DNMT3B. Then, the upregulation of BDNF expression in HPC mice was generated from a decrease in DNA methylation of the BDNF gene promoter detected by pyrophosphate sequencing. Subsequently, upregulation of BDNF activated BDNF/TrkB signaling and ultimately improved learning and spatial memory in HPC mice. Moreover, after mice were intracerebroventricularly injected with the DNMT inhibitor, the restraint of DNA methylation accompanied by an increase of BDNF and BDNF/TrkB signaling was also discovered. Finally, we observed that the inhibitor of BDNF/TrkB signaling prevented HPC from ameliorating learning and memory in mice. However, the DNMT inhibitor promoted spatial cognition in mice. Thus, we suggest that HPC may upregulate BDNF by inhibiting DNMTs and decreasing DNA methylation of the BDNF gene and then activate BDNF/TrkB signaling to improve learning and memory in mice. This may provide theoretical guidance for the clinical treatment of cognitive dysfunction caused by ischemia/hypoxia disease.

目的 通过稳定表达 70 kD热休克蛋白(HSP)4 重组蛋白(HSPA4)-增强型绿色荧光蛋白(EGFP)的SH-SY5Y细胞系,直观实时反映细胞内未折叠蛋白反应(UPR)的变化.方法 以SH-SY5Y细胞的cDNA为模板,克隆HSPA4,并连接EGFP.将HSPA4-EGFP连接入慢病毒载体中,慢病毒转染SH-SY5Y细胞.使用 3 μg/ml嘌呤霉素处理细胞 1 个月,筛选得到HSPA4-EGFP稳转系.通过实时荧光定量聚合酶链反应(QPCR)、Western印迹、共聚焦显微镜检测等方法,证实HS-PA4-EGFP稳转系能有效且实时反映UPR的变化.结果 扩增到HSPA4-EGFP,并成功构建HSPA4-EGFP稳定表达的UPR报告系统.QPCR检测表明,构建的报告系统可以指示UPR.在激光共聚焦显微镜下,可以观察到细胞内绿色EGFP的点状分布,其能够反映UPR错误折叠蛋白的多少及分布,分别加入UPR激活剂、抑制剂后,细胞内绿色EGFP的点状分布明显增多和减少,差异有统计学意义(P<0.05).Western印迹检测到细胞中HSPA4-EGFP融合蛋白表达条带.结论 成功构建HS-PA4-EGFP稳转系,能够直观实时反映UPR变化,为深入研究UPR及其相关疾病(包括老年性疾病)提供工具.

Background: 5-Aza-2 '-deoxycytidine (5-Aza-CdR) is a demethylating agent that has various biological effects related to DNA methylation. DNA methylation plays important roles in learning and memory. We have reported that 5-Aza-CdR improved the performance of mice in the water maze and step-down tests. Some behaviours have been well recognized to be mediated by neurogenesis in the hippocampus. The Notch signalling pathway plays a key role in adult hippocampal neurogenesis. In this study, we examined whether 5-Aza-CdR (DNA methyltransferase inhibitor) affects neurogenesis and Notch1 expression. Methods: The learning and memory behaviour of mice was evaluated by a conditioned avoidance learning 24 h after 5-Aza-CdR treatment. The mRNA and protein expression levels of Notch1 and HES1 were measured by real-time PCR and Western blotting. The 5-bromo-2 '-deoxyuridine (BrdU)-positive cells and the expression of Notch1 in the hippocampal DG were observed through laser confocal microscopy. To further clarify whether 5-Aza-CdR affects behaviour through neurogenesis, the expression level of Notch1, cell viability and cell cycle were analysed using the HT22 cell line. Results: The behaviour in conditioned avoidance learning was improved, while neurogenesis and the Notch1 pathway were increased in the hippocampus of mice that were injected with 5-Aza-CdR. In vitro experiments showed that 5-Aza-CdR increased the expression of the Notch1 pathway and upregulated S-phase in the cell cycle and cell viability. Conclusions: Our results suggest that the effect of 5-Aza-CdR on behaviour may be related to an increase in neurogenesis with upregulation of the Notch1 pathway in the hippocampus.

目的:研究5-氮-2'脱氧胞苷(5-Aza-cdR)通过影响DNA甲基转移酶功能,调节BDNF表达并参与学习记忆的相关机制.方法:以70只6～8周18～22克雄性ICR小鼠为研究对象,随机分为三组:注射PBS为C组,25只;注射5-Aza-cdR为注射组,30只;注射K252a为K252a注射组,15只.通过Western blot、real-time PCR、免疫荧光技术,检测小鼠海马脑区DNA甲基转移酶3B(DNMT3B)和脑源性神经营养因子(BDNF)在mRNA和蛋白水平的表达变化,通过水迷宫实验检测小鼠的学习记忆能力.结果:与C组比较,小鼠经过5-Aza-cdR处理24 h后,DNMT3B的蛋白水平和mRNA水平均有所下降(P<0.05),BDNF的蛋白和mRNA水平显著增加(P<0.05).与C组比较,5-Aza-cdR注射组的BDNF荧光强度显著增强(P<0.05),小鼠找到目标平台的潜伏期显著降低(P<0.05),目标象限百分比以及目标象限穿越次数均显著增加(P<0.05).结论:5-Aza-cdR下调DNMT3B表达后,通过上调小鼠海马中BDNF的表达,可提高小鼠的学习记忆能力.

Background:It has been reported that ischemia and ischemic preconditioning (IPC) have different effects on the expression of tuberous sclerosis complex 1 (TSC1), which may contribute to the tolerance to ischemia/hypoxia with the increase of autophagy. The mechanisms of TSC1 differential expression are still unclear under ischemia/IPC conditions in hippocampal Cornu Ammon 1 (CA1) and Cornu Ammon 3 (CA3) area neuronal cells. While we have shown that 5-Aza-CdR, a DNA methyltransferase inhibitor, can upregulate TSC1 and increase hypoxic tolerance by autophagy in vivo and in vitro, in this study, we examined whether DNA methylation was involved in the differential expression of TSC1 in the CA1 and CA3 regions induced by hypoxic preconditioning (HPC).Methods:Level of rapamycin (mTOR) autophagy, a downstream molecular pathway of TSC1/TSC2 complex, was detected in HPC mouse hippocampal CA1 and CA3 areas as well as in the HPC model of mouse hippocampal HT22 cells. DNA methylation level of TSC1 promoter (-720 bp~ -360 bp) was determined in CA1 and CA3 areas by bisulfite-modified DNA sequencing (BMDS). At the same time, autophagy was detected in HT22 cells transfected with GFP-LC3 plasmid. The role of TSC1 in neuroprotection was measured by cell viability and apoptosis, and the role of TSC1 in metabolism was checked by ATP assay and ROS assay in HT22 cells that overexpressed/knocked down TSC1.Results:HPC upregulated the expression of TSC1, downregulated the level of P-mTOR (Ser2448) and P-p70S6K (Thr389), and enhanced the activity of autophagy in both in vivo and in vitro. The increased expression of TSC1 in HPC may depend on its DNA hypomethylation in the promoter region in vivo. HPC also could reduce energy consumption in HT22 cells. Overexpression and knockdown of TSC1 can affect cell viability, cell apoptosis, and metabolism in HT22 cells exposed to hypoxia.Conclusion:TSC1 expression induced by HPC may relate to the downregulation of its DNA methylation level with the increase of autophagy and the decrease of energy demand.

目的 研究远隔缺血预适应(RIPC)血清外泌体对SH-SY5Y细胞基因表达的影响,探讨差异表达基因对神经细胞在糖氧剥夺(OGD)耐受中的作用.方法 RIPC前后分离人血清外泌体,取RIPC前外泌体(HuE-C)和RIPC后外泌体(HuE-RIPC).将SH-SY5Y细胞分为空白组(正常培养细胞),对照组(培养液加入HuE-C)和RIPC组(培养液加入HuE-RIPC),采用高通量测序各组细胞基因表达并进行生物信息学分析,将3组细胞置于OGD条件下,检测细胞活力,采用实时定量PCR及Western blot检测与神经低氧/缺血相关的基因表达.结果 生物信息学显示,RIPC组热休克蛋白(HSP)基因差异表达增加.MTS检测发现,OGD条件下,RIPC组细胞活力明显高于空白组(0.673±0.056 vs 0.481±0.027,P＜0.01).实时定量PCR和Western blot检测显示,OGD条件下,RIPC组HSP70 mRNA表达和蛋白表达明显高于空白组和对照组(P＜0.05).结论 人RIPC血清外泌体可引起众多基因表达变化,其中HSP70表达增加可能是其低氧耐受增加的一个原因.

Objective To study the effects of 5-Aza-2′-deoxycytidine (5-Aza-CdR) on learning and memory capacity of mice by step-down test and tyrosine phosphorylation of NR2B 1472 site (p-Y1472 NR2B) in hippocampus. Methods Mice were randomly divided into control group (control) and 5-Aza-CdR treatment group (5-Aza-CdR). In the test, 5-Aza-CdR (10 μmol/L) was administered to treatment group mice via intracerebroventricular injection. The ability of learning and memory in mice was evaluated by step-down test. The mRNA level of NR2B in hippocampal tissues of mice was measured by real-time PCR. NR2B and p-Y1472 NR2B in hippocampal tissues of mice were measured by Western blot and immunofluorescence. Results In the 5-Aza-CdR treatment group, the latency period was increased, the error records were decreased, the behavioral memory was significantly improved in step-down test(P＜0.05). While the expression of NR2B mRNA and protein in the hippocampus of mice was not significantly changed, the phosphorylation of p-Y1472 NR2B was increased(P＜0.05), and CDK5 activity was decreased(P＜0.05). There was no difference in NR2B fluorescence intensity between CA1 region and CA3 region, while p-Y1472 NR2B in CA1 region was significantly increased (P＜0.05). Conclusions The effects of 5-Aza-CdR on behavioral memory ability may be related to the p-Y1472 NR2B in the hippocampus of mice.

目的:研究经低氧预适应处理的血管内皮细胞分泌的物质对神经细胞低氧耐受的影响.方法:分别将人微血管内皮细胞(HMEC-1)经过低氧预适应(HPC)处理及正常培养,分离其培养基并通过滤膜过滤.人神经母细胞瘤细胞系(SH-SY5Y)细胞分为低氧预适应组(HPC组)、培养基对照组(Medium-C组)和空白对照组(SY5Y-C组),三组细胞分别加入经HPC的HMEC-1细胞的培养基、正常培养HMEC-1细胞的培养基和正常培养SH-SY5Y细胞的培养基.随后三个组细胞均给予低氧处理.使用MTS法检测神经细胞活力,Western Blot检测caspase-3蛋白表达水平.结果:SY5Y-C组、Medium-C组和HPC组的细胞活力分别是(0.9967±0.0033)、(1.0751±0.0447)和(1.4737±0.0325),HPC组细胞活力显著增加(P<0.05);三个组caspase-3蛋白表达的相对丰度分别是(0.9950±0.0050)、(0.9893±0.0787)、(0.8180±0.0361),HPC组细胞的caspase-3蛋白表达显著受到抑制(P<0.05).结论:内皮细胞在经过低氧预适应处理后分泌的物质可以增加神经细胞的低氧耐受.

目的 探讨神经生长因子(NGF)对小鼠海马神经元细胞系HT22分化及DNMT表达的影响.方法 小鼠海马神经元细胞系HT22分为未分化组(不经特殊处理)和分化组(50 ng/ml NGF-β处理细胞).通过活细胞动态成像与分析系统IncuCyte分析NGF是否参与小鼠海马神经元细胞系HT22的分化.实时定量聚合酶链反应和Western印迹法检测DNA甲基转移酶(DNMT)基因和蛋白质表达.应用甲基转移酶活性试剂盒检测DNMTs活性.应用流式细胞术检测细胞周期.结果 NGF处理后,分化组与未分化组HT22细胞相比DNMT1、DNMT3A、DNMT3B的mRNA表达水平显著升高了13倍、8倍、5倍(P<0.05),其中DNMT1、DNMT3A蛋白质表达量显著增加1.5倍(P<0.05),DNMTs活性增强(P<0.05),S期细胞百分比显著增加.结论 NGF 诱导的HT22 细胞的分化与DNMTs 的激活有关.

目的 探讨纳米氧化铈对斑马鱼胚胎早期发育的影响及机制.方法 将生长发育良好的斑马鱼胚胎随机分为对照组和纳米氧化铈染毒50、100、200、400、800 mg/L剂量组,每组40个.各剂量组斑马鱼胚胎分别予相应质量浓度的纳米氧化铈染毒5d,对照组斑马鱼胚胎不予染毒.观察斑马鱼胚胎死亡以及畸形情况,采用激光共聚焦显微镜实时记录斑马鱼胚胎心率,采用蛋白免疫印迹实验检测斑马鱼胚胎活化的含半胱氨酸天冬氨酸蛋白水解酶3(cleaved Caspase-3)、微管相关蛋白1轻链3(LC3)蛋白的表达情况.结果 各组斑马鱼胚胎死亡率、胚胎畸形率随染毒剂量增大而增加(P＜0.01);800 mg/L剂量组斑马鱼胚胎心率低对照组[(77±8)vs(93 ±4)次/分,P＜0.01].各组斑马鱼胚胎LC3-Ⅱ蛋白相对表达水平比较,差异无统计学意义(P＞0.05).各剂量组斑马鱼胚胎cleaved Caspase-3蛋白相对表达水平均高于对照组(P＜0.05);200 mg/L剂量组斑马鱼胚胎cleaved Caspase-3蛋白相对表达水平高于50 mg/L剂量组(P＜0.05).结论 纳米氧化铈颗粒可能通过激活细胞凋亡,引起斑马鱼胚胎早期发育的毒性效应.

Although of nanoparticles extensively studied, the effects on the early development are not well understood. In this study, we attempted to explain the toxic effects of neodymium oxide (Nd 2 O 3 ) nanoparticles on early development. Methods: We added the Nd 2 O 3 nanoparticles at different concentrations and recorded the mortality and malformation rate per 24 hrs under a microscope. The live embryos treated with Nd 2 O 3 nanoparticles were imaged as movies and Z step lapses with a confocal microscope, and heart rates were counted for 30 s to measure the cardiac function. The live Tg ( Flk1 :EGFP) transgenic embryos exposed to Nd 2 O 3 nanoparticles were observed under confocal microscope to measure the cerebrovascular development. Subsequently, we extracted the total protein for Western blot at 5 days post-fertilisation (dpf). Embryos were collected to undergo TUNEL staining for apoptosis detection. Results: Nd 2 O 3 nanoparticles disturbed embryo development at high concentrations (>200 μ g/mL). The mortality and malformation rate gradually increased in a dose-dependent manner by morphological observation, while the Nd 2 O 3 median lethal concentration (LD50) was 203.4 μ g/mL at 120 hrs post-fertilisation (hpf). Furthermore, the Nd 2 O 3 treated embryos showed severe arrhythmia and reduced heart rate. We also observed the markedly cerebrovascular disappearance at middle concentration (100 and 200 μ g/mL). The downregulated autophagy fl ux in brain blood vessels and increased apoptosis level in neurons might affect vessels sprouting and contribute to the vanished cerebrovascular. Conclusion: The results suggested that the embryos exposed to Nd 2 O 3 activated the apoptosis pathway and induced toxicity and abnormal cardiac/cerebrovascular development.

目的:研究低氧及低氧预适应对HT22细胞中学习记忆相关基因PP1γ和Reelin表达的影响.方法:神经细胞HT22分为对照组(C)、低氧组(H)和低氧预适应组(HPC)并给于相应处理.采用Real-time PCR及western blot方法检测PP1γ和Reelin的mRNA和蛋白表达水平.结果:低氧预适应降低PP1γ的表达,增加Reelin的表达.结论:低氧预适应可能通过调节HT22细胞PP1γ和Reelin的表达增加细胞低氧耐受.