Axonal growth inhibitors and their receptors in spinal cord injury:from biology to clinical translation

Axonal growth inhibitors are released during traumatic injuries to the adult mammalian central nervous system, including after spinal cord injury. These molecules accumulate at the injury site and form a highly inhibitory environment for axonal regeneration. Among these inhibitory molecules, myelinassociated inhibitors, including neurite outgrowth inhibitor A, oligodendrocyte myelin glycoprotein, myelin-associated glycoprotein, chondroitin sulfate proteoglycans and repulsive guidance molecule A are of particular importance. Due to their inhibitory nature, they represent exciting molecular targets to study axonal inhibition and regeneration after central injuries. These molecules are mainly produced by neurons, oligodendrocytes, and astrocytes within the scar and in its immediate vicinity. They exert their effects by binding to specific receptors, localized in the membranes of neurons. Receptors for these inhibitory cues include Nogo receptor 1, leucine-rich repeat, and Ig domain containing 1 and p75 neurotrophin receptor/tumor necrosis factor receptor superfamily member 19(that form a receptor complex that binds all myelin-associated inhibitors), and also paired immunoglobulin-like receptor B. Chondroitin sulfate proteoglycans and repulsive guidance molecule A bind to Nogo receptor 1, Nogo receptor 3, receptor protein tyrosine phosphatase σ and leucocyte common antigen related phosphatase, and neogenin, respectively. Once activated, these receptors initiate downstream signaling pathways, the most common amongst them being the Rho A/ROCK signaling pathway. These signaling cascades result in actin depolymerization, neurite outgrowth inhibition, and failure to regenerate after spinal cord injury. Currently, there are no approved pharmacological treatments to overcome spinal cord injuries other than physical rehabilitation and management of the array of symptoms brought on by spinal cord injuries. However, several novel therapies aiming to modulate these inhibitory proteins and/or their receptors are under investigation in ongoing clinical trials. Investigation has also been demonstrating that combinatorial therapies of growth inhibitors with other therapies, such as growth factors or stem-cell therapies, produce stronger results and their potential application in the clinics opens new venues in spinal cord injury treatment.

Upregulation of circ0000381 atienuates microglial/macrophage pyroptosis after spinal cord injury

Neuroinflammation exacerbates secondary damage after spinal cord injury,while microglia/macrophage pyroptosis is important to neuroinflammation.Circular RNAs(circRNAs)play a role in the central nervous system.However,the functional role and mechanism of circRNAs in regulating microglia/macrophage pyroptosis after spinal cord injury are still poorly studied.In the present study,we detected microglia/macrophage pyroptosis in a female rat model of spinal cord injury,along with upregulated levels of circ0000381 in the spinal cord.Our further experimental results suggest that circ0000381 may function as a sponge to sequester endogenous microRNA423-3p(miR-423-3p),which can increase the expression of NOD-like receptor 3(NLRP3),a pyroptosis marker.Therefore,upregulation of circ0000381 may be a compensatory change after spinal cord injury to attenuate microglia/macrophage pyroptosis.Indeed,knockdown of circ0000381 expression exacerbated microglia/macrophage pyroptosis.Collectively,our findings provide novel evidence for the upregulation of circ0000381,which may serve as a neuroprotective mechanism to attenuate microglia/macrophage pyroptosis after spinal cord injury.Accordingly,circ0000381 may be a novel therapeutic target for the treatment of spinal cord injury.

Photobiomodulation inhibits the expression of chondroitin sulfate proteoglycans after spinal cord injury via the Sox9 pathway

Both glial cells and glia scar greatly affect the development of spinal cord injury and have become hot spots in research on spinal cord injury treatment.The cellular deposition of dense extracellular matrix proteins such as chondroitin sulfate proteoglycans inside and around the glial scar is known to affect axonal growth and be a major obstacle to autogenous repair.These proteins are thus candidate targets for spinal cord injury therapy.Our previous studies demonstrated that 810 nm photo biomodulation inhibited the formation of chondroitin sulfate proteoglycans after spinal cord injury and greatly improved motor function in model animals.However,the specific mechanism and potential targets involved remain to be clarified.In this study,to investigate the therapeutic effect of photo biomodulation,we established a mouse model of spinal cord injury by T9 clamping and irradiated the injury site at a power density of 50 mW/cm~2 for 50 minutes once a day for 7 consecutive days.We found that photobiomodulation greatly restored motor function in mice and down regulated chondroitin sulfate proteoglycan expression in the injured spinal cord.Bioinformatics analysis revealed that photobiomodulation inhibited the expression of proteoglycan-related genes induced by spinal cord injury,and versican,a type of proteoglycan,was one of the most markedly changed molecules.Immunofluorescence staining showed that after spinal cord injury,versican was present in astrocytes in spinal cord tissue.The expression of versican in primary astrocytes cultured in vitro increased after inflammation induction,whereas photobiomodulation inhibited the expression of ve rsican.Furthermore,we found that the increased levels of p-Smad3,p-P38 and p-Erk in inflammatory astrocytes were reduced after photobiomodulation treatment and after delivery of inhibitors including FR 180204,(E)-SIS3,and SB 202190.This suggests that Sma d 3/Sox9 and MAP K/Sox9 pathways may be involved in the effects of photobiomodulation.In summary,our findings show that photobiomodulation modulates the expression of chondroitin sulfate proteoglycans,and versican is one of the key target molecules of photo biomodulation.MAPK/Sox9 and Smad3/Sox9 pathways may play a role in the effects of photo biomodulation on chondroitin sulfate proteoglycan accumulation after spinal cord injury.

Deep Transfers of p-Class Tower Groups

Let p be a prime. For any finite p-group G, the deep transfers T H,G ' : H / H ' → G ' / G " from the maximal subgroups H of index (G:H) = p in G to the derived subgroup G ' are introduced as an innovative tool for identifying G uniquely by means of the family of kernels ùd(G) =(ker(T H,G ')) (G: H) = p. For all finite 3-groups G of coclass cc(G) = 1, the family ùd(G) is determined explicitly. The results are applied to the Galois groups G =Gal(F3 (∞)/ F) of the Hilbert 3-class towers of all real quadratic fields F = Q(√d) with fundamental discriminants d > 1, 3-class group Cl3(F) □ C3 × C3, and total 3-principalization in each of their four unramified cyclic cubic extensions E/F. A systematic statistical evaluation is given for the complete range 1 d 7, and a few exceptional cases are pointed out for 1 d 8.

Dual-targeting AAV9P1-mediated neuronal reprogramming in a mouse model of traumatic brain injury

Traumatic brain injury results in neuronal loss and glial scar formation.Replenishing neurons and eliminating the consequences of glial scar formation are essential for treating traumatic brain injury.Neuronal reprogramming is a promising strategy to convert glial scars to neural tissue.However,previous studies have reported inconsistent results.In this study,an AAV9P1 vector incorporating an astrocyte-targeting P1 peptide and glial fibrillary acidic protein promoter was used to achieve dual-targeting of astrocytes and the glial scar while minimizing off-target effects.The results demonstrate that AAV9P1 provides high selectivity of astrocytes and reactive astrocytes.Moreover,neuronal reprogramming was induced by downregulating the polypyrimidine tract-binding protein 1 gene via systemic administration of AAV9P1 in a mouse model of traumatic brain injury.In summary,this approach provides an improved gene delivery vehicle to study neuronal programming and evidence of its applications for traumatic brain injury.

Exosomes derived from microglia overexpressing miR-124-3p alleviate neuronal endoplasmic reticulum stress damage after repetitive mild traumatic brain injury

We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repetitive mild traumatic brain injury remains unclear.In this study,we first used an HT22 scratch injury model to mimic traumatic brain injury,then co-cultured the HT22 cells with BV2 microglia expressing high levels of miR-124-3p.We found that exosomes containing high levels of miR-124-3p attenuated apoptosis and endoplasmic reticulum stress.Furthermore,luciferase reporter assay analysis confirmed that miR-124-3p bound specifically to the endoplasmic reticulum stress-related protein IRE1α,while an IRE1αfunctional salvage experiment confirmed that miR-124-3p targeted IRE1αand reduced its expression,thereby inhibiting endoplasmic reticulum stress in injured neurons.Finally,we delivered microglia-derived exosomes containing miR-124-3p intranasally to a mouse model of repetitive mild traumatic brain injury and found that endoplasmic reticulum stress and apoptosis levels in hippocampal neurons were significantly reduced.These findings suggest that,after repetitive mild traumatic brain injury,miR-124-3 can be transferred from microglia-derived exosomes to injured neurons,where it exerts a neuroprotective effect by inhibiting endoplasmic reticulum stress.Therefore,microglia-derived exosomes containing miR-124-3p may represent a novel therapeutic strategy for repetitive mild traumatic brain injury.

TCP1表达对HL60和HL60/A 细胞增殖及细胞内药物蓄积的调控作用及其机制

目的:研究TCP1表达对HL60及HL60/A细胞增殖及细胞内药物蓄积的影响及其机制。方法:利用慢病毒转染技术构建敲低、过表达TCP1的HL60/A细胞和HL60细胞及其对照组细胞,Western blot评估敲低、过表达效率。采用CCK-8法检测细胞增殖能力;激光共聚焦显微镜及流式细胞术检测细胞内的药物蓄积;流式细胞术及Western blot检测膜转运蛋白(MRP1、P-gP)及p-AKT的表达水平。结果:在HL60/A细胞中敲低TCP1的表达能够抑制细胞增殖,增加细胞内药物蓄积,降低转运蛋白MRP1及P-gP的表达,在HL60细胞中过表达TCP1能够促进细胞的增殖,减少细胞内药物蓄积,提高转运蛋白MRPI及P-gP的表达。同时利用PI3K抑制剂LY294002抑制PI3K/AKT信号能够拮抗TCP1过表达所致的细胞增殖活性增强、细胞内药物蓄积减少及MRP1、P-gP表达的升高。结论:TCP1能够促进细胞增殖,并通过激活PI3K/AKT信号促进转运蛋白MRP1、P-gP的表达,降低细胞内的药物蓄积。

Updates on management of gliomas in the molecular age

Gliomas are primary brain tumors derived from glial cells of the central nervous system,afflicting both adults and children with distinct characteristics and therapeutic challenges.Recent developments have ushered in novel clinical and molecular prognostic factors,reshaping treatment paradigms based on classi-fication and grading,determined by histological attributes and cellular lineage.This review article delves into the diverse treatment modalities tailored to the specific grades and molecular classifications of gliomas that are currently being discussed and used clinically in the year 2023.For adults,the therapeutic triad typically consists of surgical resection,chemotherapy,and radiotherapy.In contrast,pediatric gliomas,due to their diversity,require a more tailored approach.Although complete tumor excision can be curative based on the location and grade of the glioma,certain non-resectable cases demand a chemotherapy approach usually involving,vincristine and carboplatin.Addi-tionally,if surgery or chemotherapy strategies are unsuccessful,Vinblastine can be used.Despite recent advancements in treatment methodologies,there remains a need of exploration in the literature,particularly concerning the efficacy of treatment regimens for isocitrate dehydrogenase type mutant astrocytomas and fine-tuned therapeutic approaches tailored for pediatric cohorts.This review article explores into the therapeutic modalities employed for both adult and pediatric gliomas in the context of their molecular classification.

Soluble p75 neurotrophic receptor as a reliable biomarker in neurodegenerative diseases: what is the evidence?

Neurodegenerative diseases are often misdiagnosed,especially when the diagnosis is based solely on clinical symptoms.The p75 neurotrophic receptor(p75^(NTR))has been studied as an index of sensory and motor nerve development and maturation.Its cleavable extracellular domain(ECD)is readily detectable in various biological fluids including plasma,serum and urine.There is evidence for increased p75NTR ECD levels in neurodegenerative diseases such as Alzheimer’s disease,amyotrophic lateral sclerosis,age-related dementia,schizophrenia,and diabetic neuropathy.Whether p75^(NTR) ECD could be used as a biomarker for diagnosis and/or prognosis in these disorders,and whether it could potentially lead to the development of targeted therapies,remains an open question.In this review,we present and discuss published studies that have evaluated the relevance of this emerging biomarker in the context of various neurodegenerative diseases.We also highlight areas that require further investigation to better understand the role of p75^(NTR) ECD in the clinical diagnosis and management of neurodegenerative disorders.

Exercise-induced modulation of miR-149-5p and MMP9 in LPS-triggered diabetic myoblast ER stress: licorice glycoside E as a potential therapeutic target

Background:This study explores the relationship between endoplasmic reticulum(ER)stress and diabetes,particularly focusing on the impact of physical exercise on ER stress mechanisms and identifying potential therapeutic drugs and targets for diabetes-related sepsis.The research also incorporates traditional physical therapy perspectives,emphasizing the genomic insights gained from exercise therapy in disease management and prevention.Methods:Gene analysis was conducted on the GSE168796 and GSE94717 datasets to identify ER stress-related genes.Gene interactions and immune cell correlations were mapped using GeneCard and STRING databases.A screening of 2,456 compounds from the TCMSP database was performed to identify potential therapeutic agents,with a focus on their docking potential.Techniques such as luciferase reporter gene assay and RNA interference were used to examine the interactions between microRNA-149-5p and MMP9.Results:The study identified 2,006 differentially expressed genes and 616 miRNAs.Key genes like MMP9,TNF-α,and IL1B were linked to an immunosuppressive state.Licorice glycoside E demonstrated high affinity for MMP9,suggesting its potential effectiveness in treating diabetes.The constructed miRNA network highlighted the regulatory roles of MMP9,IL1B,IFNG,and TNF-α.Experimental evidence confirmed the binding of microRNA-149-5p to MMP9,impacting apoptosis in diabetic cells.Conclusion:The findings highlight the regulatory role of microRNA-149-5p in managing MMP9,a crucial gene in diabetes pathophysiology.Licorice glycoside E emerges as a promising treatment option for diabetes,especially targeting MMP9 affected by ER stress.The study also underscores the significance of physical exercise in modulating ER stress pathways in diabetes management,bridging traditional physical therapy and modern scientific understanding.Our study has limitations.It focuses on the microRNA-149-5p-MMP9 network in sepsis,using cell-based methods without animal or clinical trials.Despite strong in vitro findings,in vivo studies are needed to confirm licorice glycoside E’s therapeutic potential and understand the microRNA-149-5p-MMP9 dynamics in real conditions.

Homer1a reduces inflammatory response after retinal ischemia/reperfusion injury

Elevated intraocular pressure(IOP)is one of the causes of retinal ischemia/reperfusion injury,which results in NRP3 inflammasome activation and leads to visual damage.Homerla is repo rted to play a protective role in neuroinflammation in the cerebrum.However,the effects of Homerla on NLRP3inflammasomes in retinal ischemia/reperfusion injury caused by elevated IOP remain unknown.In our study,animal models we re constructed using C57BL/6J and Homer1^(flox/-)/Homerla^(+/-)/Nestin-Cre^(+/-)mice with elevated IOP-induced retinal ischemia/repe rfusion injury.For in vitro expe riments,the oxygen-glucose deprivation/repe rfusion injury model was constructed with M uller cells.We found that Homerla ove rexpression amelio rated the decreases in retinal thickness and Muller cell viability after ischemia/reperfusion injury.Furthermore,Homerla knockdown promoted NF-κB P65^(Ser536)activation via caspase-8,NF-κB P65 nuclear translocation,NLRP3 inflammasome formation,and the production and processing of interleukin-1βand inte rleukin-18.The opposite results we re observed with Homerla ove rexpression.Finally,the combined administration of Homerla protein and JSH-23 significantly inhibited the reduction in retinal thickness in Homer1^(flox/-)Homer1a^(+/-)/Nestin-Cre^(+/-)mice and apoptosis in M uller cells after ischemia/reperfusion injury.Taken together,these studies demonstrate that Homer1a exerts protective effects on retinal tissue and M uller cells via the caspase-8/NF-KB P65/NLRP3 pathway after I/R injury.

黄芩苷对干酵母致热大鼠的解热作用及血清TNF-α、IL-1β、IL-6、PGE_(2)、cAMP和脑组织NF-κB表达的影响

目的:观察黄芩苷对干酵母致热大鼠的解热作用并探讨其作用机制。方法:采用背部皮下注射干酵母构建大鼠发热模型,SD雄性大鼠随机分为正常对照,模型组,阳性组(阿司匹林,0.1 g/kg),黄芩苷高、中、低剂量组(160、80、40 mg/kg),连续给药3 d,测定各组大鼠肛温的变化;酶联免疫法(ELISA)检测血清肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白细胞介素-6(IL-6)、前列腺素E_(2)(PGE_(2))与环磷酸腺苷(cAMP)水平;Western Blot检测各组大鼠脑组织NF-κB p65(核转录因子-κB p65)蛋白表达。结果:黄芩苷高剂量组有显著解热效果(P<0.01),黄芩苷各剂量组均可不同程度降低大鼠血清TNF-α、IL-1β、IL-6、PGE 2和cAMP含量;与正常组比较,模型组脑组织NF-κB p65蛋白表达增多,黄芩苷高剂量组可明显降低大鼠脑组织NF-κB p65表达(P<0.05)。结论:黄芩苷可显著性降低干酵母引起的体温升高,解热机制可能与抑制TNF-α、IL-1β、IL-6、PGE_(2)与cAMP的分泌和减少脑组织NF-κB p65蛋白表达有关。

针刺对卵巢储备功能减退大鼠血清激素及PI3K/Akt/mTOR信号通路蛋白表达的影响

目的:观察针刺对卵巢储备功能减退(DOR)大鼠血清激素、卵巢形态学变化及卵巢组织中p-PI3K、p-Akt及p-mTOR蛋白表达的影响,探索针刺治疗DOR的可能机制。方法:18只SPF级SD雌性大鼠随机分为空白组、模型组和针刺组,每组6只;模型组、针刺组采用腹腔注射环磷酰胺制备DOR大鼠模型,空白组仅腹腔注射同等量生理盐水。造模后针刺组取肾俞、关元、中极、气海、太溪、双侧三阴交和双侧子宫穴进行针刺,每次留针20 min,1次/d,连续21 d。空白组、模型组:仅抓取不进行干预,1次/d,连续21 d。观察比较各组间大鼠的血清激素改变情况,HE染色法观察大鼠卵巢形态学改变、卵巢中各级卵泡及黄体的数目变化,颗粒细胞层厚度改变;免疫组织化学法、Western blot法检测卵巢组织中p-PI3K、p-Akt及p-mTOR的蛋白表达。结果:与空白组比较,模型组血清FSH、LH水平升高(P<0.05),血清E2、AMH水平降低,差异具有统计学意义(P<0.05);卵巢组织中各级卵泡数明显减少,差异具有统计学意义(P<0.05);卵巢组织中PI3K、Akt及mTOR蛋白表达水平升高,差异具有统计学意义(P<0.05)。与模型组比较,针刺组血清FSH、LH水平显著降低,差异具有统计学意义(P<0.05);血清E2、AMH水平显著升高,差异具有统计学意义(P<0.05);卵巢组织中各级卵泡数明显增多,差异具有统计学意义(P<0.05);卵巢组织中p-PI3K、p-Akt及p-mTOR蛋白表达水平降低,差异具有统计学意义(P<0.05)。结论:针刺可以有效改善DOR大鼠卵巢储备功能,促进卵泡数量增加。作用机制可能与针刺调节卵巢组织中p-PI3K、p-Akt及p-mTOR蛋白表达有关。

Effects of Inoculation with Phosphate Solubilizing Bacteria on the Physiology,Biochemistry,and Expression of Genes Related to the Protective Enzyme System of Fritillaria taipaiensis P.Y.Li

Fritillaria taipaiensis P.Y.Li is a widely used medicinal herb in treating pulmonary diseases.In recent years,its wild resources have become scarce,and the demand for efficient artificial cultivation has significantly increased.This article is the first to apply phosphate solubilizing bacteria isolated from the rhizosphere soil of F.taipaiensis P.Y.Li to the cultivation process of F.taipaiensis P.Y.Li.The aim is to identify suitable reference strains for the artificial cultivation and industrial development of F.taipaiensis P.Y.Li by examining the effects of various phosphate solubilizing bacteria and their combinations on photosynthesis,physiological and biochemical properties,and gene expression related to the protective enzyme system in F.taipaiensis P.Y.Li.The experiment,conducted in pots at room temperature,included a control group(CK)and groups inoculated with inorganic phosphorussolubilizing bacteria:W1(Bacillus cereus),W2(Serratia plymuthica),W12(Bacillus cereus and Serratia plymuthica),and groups inoculated with organophosphorus-solubilizing bacteria:Y1(Bacillus cereus),Y2(Bacillus cereus),Y12(Bacillus cereus and Bacillus cereus),totaling seven groups.Compared to CK,most growth indices in the bacterial addition groups showed significant differences,with W12 achieving the highest values in all indices except the leaf area index.The content of photosynthetic pigments,photosynthetic parameters,and osmoregulatory substances increased variably in each bacterial treatment group.W12 exhibited the highest content of chlorophyll a and soluble protein,while W1 had the highest free proline content.The activities of peroxidase(POD),superoxide dismutase(SOD),and catalase(CAT)in all inoculated groups were higher than in CK,with significant changes in SOD and CAT activities.The malondialdehyde(MDA)content in all inoculated groups was lower than in CK,with Y12 being the lowest,at approximately 30%of CK.Gene expression corresponding to these three enzymes also increased variably,with POD expression in Y2 being the highest at 2.73 times that of CK.SOD and CAT expression in Y12 were the highest,at 1.84 and 4.39 times that of CK,respectively.These results indicate that inoculating phosphate solubilizing bacteria can enhance the growth of F.taipaiensis P.Y.Li,with the mixed inoculation groups W12 and Y12 demonstrating superior effects.This lays a theoretical foundation for selecting bacterial fertilizers in the cultivation process of F.taipaiensis P.Y.Li.

miR-27-3p、miR-128-3p及miR-140-3p表达在急性脑梗死患者诊疗中的价值研究

目的 探讨miR-27-3p、miR-128-3p及miR-140-3p表达在急性脑梗死(ACI)患者诊疗中的应用价值。方法 选取2020年1月至2022年12月南通市第三人民医院全科医学科收治的ACI患者100例作为研究对象,定义为ACI组,根据ACI患者3个月改良Rankin评分量表(mRS评分)将其分为预后良好组(mRS评分≤2分)68例与预后不良组(mRS>2分)32例,另选同期于南通市第三人民医院全科医学科体检的健康者100例作为对照组。记录各组miR-27-3p、miR-128-3p及miR-140-3p表达水平并进行比较,采用Pearson相关分析探讨两变量的相关性,通过绘制受试者工作特征(ROC)曲线评价miR-27-3p、miR-128-3p及miR-140-3p联合检测在ACI诊断中的效能,采用多因素Logistic回归分析探讨ACI预后的危险因素。结果 ACI组平均年龄、吸烟史比例、合并高血压比例、miR-140-3p与miR-128-3p及miR-27-3p表达水平均明显高于对照组(均P<0.05)。miR-27-3p、miR-128-3p及miR-140-3p联合检测用于诊断ACI的曲线下面积(AUC值)为0.965,敏感度为99.00%,特异度为98.00%,表明miR-27-3p、miR-128-3p及miR-140-3p联合检测用于诊断ACI的效能更高。预后不良组平均年龄、入院时NIHSS评分、梗死体积、吸烟史比例、合并高血压比例、miR-27-3p、miR-128-3p、miR-140-3p表达水平及3个月mRS评分均明显高于预后良好组(均P<0.05)。经Pearson相关分析表明ACI患者miR-27-3p、miR-128-3p、miR-140-3p表达水平均分别与入院时NIHSS评分、梗死体积、3个月mRS评分呈正相关(均P<0.05)。经多因素Logistic回归分析表明年龄(OR:1.546;95%CI:1.124~1.969)、入院时NIHSS评分(OR:1.721;95%CI:1.229~2.308)、梗死体积(OR:1.853;95%CI:1.436~2.786)、吸烟史(OR:1.517;95%CI:1.168~1.825)、miR-27-3p (OR:2.481;95%CI:1.452~3.201)、miR-128-3p (OR:1.692;95%CI:1.288-2.431)及miR-140-3p (OR:2.032;95%CI:1.141~1.976)均为ACI患者预后的独立危险因素(均P<0.05)。结论 ACI患者miR-27-3p、miR-128-3p及miR-140-3p表达水平均明显升高,通过三指标联合检测可提高对ACI的诊断效能,而预后不良ACI患者miR-27-3p、miR-128-3p及miR-140-3p表达水平则升高更为明显,并分别与入院时NIHSS评分、梗死体积、3个月mRS评分呈正相关,且miR-27-3p、miR-128-3p及miR-140-3p均为ACI患者预后的独立危险因素,应予以重点关注。

MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons

Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.

P7C3-A20 treats traumatic brain injury in rats by inhibiting excessive autophagy and apoptosis

Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various diseases,including ischemic stroke and neurodegenerative diseases.However,whether P7C3-A20 has a therapeutic effect on traumatic brain injury and its possible molecular mechanisms are unclear.Therefore,in the present study,we investigated the therapeutic effects of P7C3-A20 on traumatic brain injury and explored the putative underlying molecular mechanisms.We established a traumatic brain injury rat model using a modified weight drop method.P7C3-A20 or vehicle was injected intraperitoneally after traumatic brain injury.Severe neurological deficits were found in rats after traumatic brain injury,with deterioration in balance,walking function,and learning memory.Furthermore,hematoxylin and eosin staining showed significant neuronal cell damage,while terminal deoxynucleotidyl transferase mediated dUTP nick end labeling staining indicated a high rate of apoptosis.The presence of autolysosomes was observed using transmission electron microscope.P7C3-A20 treatment reversed these pathological features.Western blotting showed that P7C3-A20 treatment reduced microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)autophagy protein,apoptosis-related proteins(namely,Bcl-2/adenovirus E1B 19-kDa-interacting protein 3[BNIP3],and Bcl-2 associated x protein[Bax]),and elevated ubiquitin-binding protein p62(p62)autophagy protein expression.Thus,P7C3-A20 can treat traumatic brain injury in rats by inhibiting excessive autophagy and apoptosis.

黄芪甲苷经由miR-125a-5p/NLRP1轴减轻椎间盘突出髓核细胞损伤

目的在白介素-1β(IL-1β)诱导退变的人髓核细胞中,探究黄芪甲苷对miR-125a-5p及其靶基因介导的信号通路的作用,揭示黄芪甲苷(astragaloside IV,AS-IV)对髓核细胞增殖、凋亡以及炎症反应的影响。方法实时荧光定量PCR(RT-qPCR)检测miR-125a-5p和核苷酸寡聚化结构域(NOD)样受体蛋白1(nucleotide oligomerization domain(NOD)-like receptor protein 1,NLRP1)在椎间盘突出患者髓核组织中的表达,用IL-1β诱导髓核细胞退变,在20、50和80μg·mL^(-1)黄芪甲苷干预浓度下检测miR-125a-5p和NLRP1的表达,在IL-1β和80μg·mL^(-1)黄芪甲苷处理的髓核细胞中单独或共同转染miR-125a-5p模拟物和NLRP1过表达质粒,然后分别检测细胞增殖、凋亡和炎症因子分泌情况,蛋白质免疫印迹(Western blot)检测细胞中信号通路相关蛋白p56和p38的磷酸化水平。结果椎间盘突出(lumbar disc herniation,LDH)患者的髓核组织中miR-125a-5p表达下调,NLRP1表达上调。黄芪甲苷促进IL-1β处理的髓核细胞中miR-125a-5p表达,减少NLRP1表达以及p56和p38蛋白的磷酸化水平。黄芪甲苷促进髓核细胞增殖,减少凋亡和炎症反应。结论黄芪甲苷通过上调miR-125a-5p表达,抑制NLRP1的表达和NF-κB/MAPK信号通路,减少IL-1β诱导的髓核细胞损伤。

BMSC-derived Exosomes Ameliorate Peritoneal Dialysis-associated Peritoneal Fibrosis via the Mir-27a-3p/TP53 Pathway

Objective:Peritoneal fibrosis(PF)is the main cause of declining efficiency and ultrafiltration failure of the peritoneum,which restricts the long-term application of peritoneal dialysis(PD).This study aimed to investigate the therapeutic effects and mechanisms of bone marrow mesenchymal stem cells-derived exosomes(BMSC-Exos)on PF in response to PD.Methods:Small RNA sequencing analysis of BMSC-Exos was performed by second-generation sequencing.C57BL/6J mice were infused with 4.25%glucose-based peritoneal dialysis fluid(PDF)for 6 consecutive weeks to establish a PF model.A total of 36 mice were randomly divided into 6 groups:control group,1.5%PDF group,2.5%PDF group,4.25%PDF group,BMSC-Exos treatment group,and BMSC-Exos+TP53 treatment group.Reverse transcription quantitative polymerase chain reaction(RT-qPCR)was performed to measure the expression level of miR-27a-3p in BMSC-Exos and peritoneum of mice treated with different concentrations of PDF.HE and Masson staining were performed to evaluate the extent of PF.The therapeutic potential of BMSC-Exos for PF was examined through pathological examination,RT-qPCR,Western blotting,and peritoneal function analyses.Epithelial-mesenchymal transition(EMT)of HMrSV5 was induced with 4.25%PDF.Cells were divided into control group,4.25%PDF group,BMSC-Exos treatment group,and BMSC-Exos+TP53 treatment group.Cell Counting Kit-8 assay was used to measure cell viability,and transwell migration assay was used to verify the capacity of BMSC-Exos to inhibit EMT in HMrSV5 cells.Results:Small RNA sequencing analysis showed that miR-27a-3p was highly expressed in BMSC-derived exosomes compared to BMSCs.The RT-qPCR results showed that the expression of miR-27a-3p was upregulated in BMSC-Exos,but decreased in PD mice.We found that PF was glucose concentration-dependently enhanced in the peritoneum of the PD mice.Compared with the control mice,the PD mice showed high solute transport and decreased ultrafiltration volume as well as an obvious fibroproliferative response,with markedly increased peritoneal thickness and higher expression ofα-SMA,collagen-I,fibronectin,and ECM1.The mice with PD showed decreased miR-27a-3p.Peritoneal structural and functional damage was significantly attenuated after BMSC-Exos treatment,while PF and mesothelial damage were significantly ameliorated.Additionally,markers of fibrosis(α-SMA,collagen-I,fibronectin,ECM1)and profibrotic cytokines(TGF-β1,PDGF)were downregulated at the mRNA and protein levels after BMSC-Exos treatment.In HMrSV5 cells,BMSC-Exos reversed the decrease in cell viability and the increase in cell migratory capacity caused by high-glucose PDF.Western blotting and RT-qPCR analysis revealed that BMSC-Exos treatment resulted in increased expression of E-cadherin(epithelial marker)and decreased expression ofα-SMA,Snail,and vimentin(mesenchymal markers)compared to those of the 4.25%PDF-treated cells.Importantly,a dual-luciferase reporter assay showed that TP53 was a target gene of miR-27a-3p.TP53 overexpression significantly reversed the decreases in PF and EMT progression induced by BMSC-Exos.Conclusion:The present results demonstrate that BMSC-Exos showed an obvious protective effect on PD-related PF and suggest that BMSC-derived exosomal miR-27a-3p may exert its inhibitory effect on PF and EMT progression by targeting TP53.

Developing selective PI3K degraders to modulate both kinase and non-kinase functions

For the first time,proteolysis-targeting chimeras(PROTAC)technology was utilized to achieve the isoform-selective degradation of class I phosphoinositide 3-kinases(PI3Ks)in this study.Through screening and optimization,the PROTAC molecule ZM-PI05 was identified as a selective degrader of p110αin multiple breast cancer cells.More importantly,the degrader can down-regulate p85 regulatory subunit simultaneously,thereby inhibiting the non-enzymatic functions of PI3K that are independent on p110catalytic subunits.Therefore,compared with PI3K inhibitor copanlisib,ZM-PI05 displayed the stronger anti-proliferative activity on breast cancer cells.In brief,a selective and efficient PROTAC molecule was developed to induce the degradation of p110αand concurrent reduction of p85 proteins,providing a tool compound for the biological study of PI3K-αby blocking its enzymatic and non-enzymatic functions.