Overexpression of low-density lipoprotein receptor prevents neurotoxic polarization of astrocytes via inhibiting NLRP3 inflammasome activation in experimental ischemic stroke

Neurotoxic astrocytes are a promising therapeutic target for the attenuation of cerebral ischemia/reperfusion injury.Low-density lipoprotein receptor,a classic cholesterol regulatory receptor,has been found to inhibit NLR family pyrin domain containing protein 3(NLRP3)inflammasome activation in neurons following ischemic stroke and to suppress the activation of microglia and astrocytes in individuals with Alzheimer’s disease.However,little is known about the effects of low-density lipoprotein receptor on astrocytic activation in ischemic stroke.To address this issue in the present study,we examined the mechanisms by which low-density lipoprotein receptor regulates astrocytic polarization in ischemic stroke models.First,we examined low-density lipoprotein receptor expression in astrocytes via immunofluorescence staining and western blotting analysis.We observed significant downregulation of low-density lipoprotein receptor following middle cerebral artery occlusion reperfusion and oxygen-glucose deprivation/reoxygenation.Second,we induced the astrocyte-specific overexpression of low-density lipoprotein receptor using astrocyte-specific adeno-associated virus.Low-density lipoprotein receptor overexpression in astrocytes improved neurological outcomes in middle cerebral artery occlusion mice and reversed neurotoxic astrocytes to create a neuroprotective phenotype.Finally,we found that the overexpression of low-density lipoprotein receptor inhibited NLRP3 inflammasome activation in oxygen-glucose deprivation/reoxygenation injured astrocytes and that the addition of nigericin,an NLRP3 agonist,restored the neurotoxic astrocyte phenotype.These findings suggest that low-density lipoprotein receptor could inhibit the NLRP3-meidiated neurotoxic polarization of astrocytes and that increasing low-density lipoprotein receptor in astrocytes might represent a novel strategy for treating cerebral ischemic stroke.

Cortico-striatal gamma oscillations are modulated by dopamine D3 receptors in dyskinetic rats

Long-term levodopa administration can lead to the development of levodopa-induced dyskinesia.Gamma oscillations are a widely recognized hallmark of abnormal neural electrical activity in levodopa-induced dyskinesia.Currently,studies have reported increased oscillation power in cases of levodopa-induced dyskinesia.However,little is known about how the other electrophysiological parameters of gamma oscillations are altered in levodopa-induced dyskinesia.Furthermore,the role of the dopamine D3 receptor,which is implicated in levodopa-induced dyskinesia,in movement disorder-related changes in neural oscillations is unclear.We found that the cortico-striatal functional connectivity of beta oscillations was enhanced in a model of Parkinson’s disease.Furthermore,levodopa application enhanced cortical gamma oscillations in cortico-striatal projections and cortical gamma aperiodic components,as well as bidirectional primary motor cortex(M1)↔dorsolateral striatum gamma flow.Administration of PD128907(a selective dopamine D3 receptor agonist)induced dyskinesia and excessive gamma oscillations with a bidirectional M1↔dorsolateral striatum flow.However,administration of PG01037(a selective dopamine D3 receptor antagonist)attenuated dyskinesia,suppressed gamma oscillations and cortical gamma aperiodic components,and decreased gamma causality in the M1→dorsolateral striatum direction.These findings suggest that the dopamine D3 receptor plays a role in dyskinesia-related oscillatory activity,and that it has potential as a therapeutic target for levodopa-induced dyskinesia.

Nuclear receptors and pathogenesis of pancreatic cancer

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with a median overall survival time of 5 mo and the five years survival less than 5%, a rate essentially unchanged over the course of the years. A well defined progression model of accumulation of genetic alterations ranging from single point mutations to gross chromosomal abnormalities has been introduced to describe the origin of this disease. However, due to the its subtle nature and concurring events PDAC cure remains elusive. Nuclear receptors (NR) are members of a large superfamily of evolutionarily conserved ligand-regulated DNA-binding transcription factors functionally involved in important cellular functions ranging from regulation of metabolism, to growth and development. Given the nature of their ligands, NR are very tempting drug targets and their pharmacological modulation has been widely exploited for the treatment of metabolic and inflammatory diseases. There are now clear evidences that both classical ligand-activated and orphan NR are involved in the pathogenesis of PDAC from its very early stages; nonetheless many aspects of their role are not fully understood. The purpose of this review is to highlight the striking connections that link peroxisome proliferator activated receptors, retinoic acid receptors, retinoid X receptor, androgen receptor, estrogen receptors and the orphan NR Nur, chicken ovalbumin upstream promoter transcription factor II and the liver receptor homologue-1 receptor to PDAC development, connections that could lead to the identification of novel therapies for this disease.

Melanocortin 3,5 receptors immunohistochemical expression in colonic mucosa of inflammatory bowel disease patients:A matter of disease activity?

BACKGROUND Melanocortin 3 and 5 receptors(i.e.,MC3R and MC5R)belong to the melanocortin family.However,data regarding their role in inflammatory bowel diseases(IBD)are currently unavailable.AIM This study aims to ascertain their expression profiles in the colonic mucosa of Crohn’s disease(CD)and ulcerative colitis(UC),aligning them with IBD disease endoscopic and histologic activity.METHODS Colonic mucosal biopsies from CD/UC patients were sampled,and immunohisto-chemical analyses were conducted to evaluate the expression of MC3R and MC5R.Colonic sampling was performed on both traits with endoscopic scores(Mayo endoscopic score and CD endoscopic index of severity)consistent with inflamed mucosa and not consistent with disease activity(i.e.,normal appearing mucosa).RESULTS In both CD and UC inflamed mucosa,MC3R(CD:+7.7 fold vs normal mucosa,P<0.01;UC:+12 fold vs normal mucosa,P<0.01)and MC5R(CD:+5.5 fold vs normal mucosa,P<0.01;UC:+8.1 fold vs normal mucosa,P<0.01)were significantly more expressed compared to normal mucosa.CONCLUSION MC3R and MC5R are expressed in the colon of IBD patients.Furthermore,expression may differ according to disease endoscopic activity,with a higher degree of expression in the traits affected by disease activity in both CD and UC,suggesting a potential use of these receptors in IBD pharmacology.

Notch3/DLL4信号通路对颈部淋巴水囊瘤后淋巴管发育的影响

目的:探究Notch3/DLL4信号通路对颈部淋巴水囊瘤(CH)后淋巴管成熟发育的影响。方法:选取C57BL/6小鼠和C57BL/6背景Notch3基因敲入小鼠,将两种小鼠分为正常组(对照组)和Notch3基因敲入组(突变组)。选取F2代小鼠处死,取小鼠颈部淋巴结分别进行HE染色、Masson染色、RT-qPCR、Western blot、免疫组化检测。结果:HE及Masson染色结果显示,与对照组比,突变组小鼠的淋巴结构被显著破坏,细胞纤维化严重。RT-qPCR结果显示,与对照组比,突变组小鼠淋巴结中DLL4、Hes1和VEGFR3 mRNA表达显著下降(P<0.05)。Western blot结果显示,与对照组比,突变组小鼠淋巴结中DLL4、Hes1和VEGFR3蛋白表达显著下降(P<0.05)。免疫组化结果显示,与对照组相比,突变组小鼠淋巴结中DLL4、Hes1、Ang2、VEGFA和VEGFR3染色较浅,蛋白表达水平下降(P<0.05)。结论:Notch3/DLL4信号通路能够调控小鼠颈部CH的形成,其机制可能与淋巴管发育有关。

NOTCH3基因C260S位点突变导致CADASIL的临床和影像学特征分析

目的探讨NOTCH3基因第5外显子C260S位点突变导致的伴有皮层下梗死和白质脑病的常染色体显性遗传性脑动脉病(cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy,CADASIL)家系的临床和影像学特征。方法选取2021年12月首都医科大学附属北京同仁医院来自同一家庭的CADASIL患者,对所有患者进行NOTCH3基因测序,回顾性分析患者的临床表现和头颅影像学特征。复习既往文献报道的导致同一位置氨基酸改变的其他突变类型的临床及影像学特征。结果4名家庭成员中,包括先证者(46岁,女)及其两个姐姐(分别为48岁和50岁)和女儿(18岁)。先证者及其父亲、两个姐姐都有偏头痛病史,其中大姐有记忆力减退;先证者患有脑梗死及伴有视觉先兆的偏头痛;先证者女儿体健;先证者父亲因脑梗死去世。4名家庭成员均存在C260S位点的NOTCH3基因突变。既往文献无此位点突变的报道,先证者头颅MRI示右侧脑桥亚急性梗死,颞叶、脑室周围及脑干异常高信号改变,其大姐脑桥可见腔隙性梗死灶。结论NOTCH3基因第5外显子c.778T>A(p.C260S)的罕见突变导致的CADASIL发病时间早,早期会出现认知障碍。合并偏头痛的脑干梗死患者,需警惕CADASIL的可能。

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.

腔隙性梗死连续住院患者NOTCH3基因突变分析

目的研究腔隙性梗死住院患者NOTCH3基因突变情况。方法纳入2020-10—2021-10广东省中医院脑病中心连续住院的腔隙性梗死患者330例,对所有患者进行NOTCH3基因突变分析,并比较基因突变阳性(CADASIL组)和阴性(非CADASIL组)腔隙性梗死患者的年龄、性别、家族史、血管危险因素、临床和影像表现等。结果330例患者中57例携带NOTCH3基因突变,其中15例为致病性突变(均为c.1630C>T;p.Arg544Cys),总体致病突变率为4.5%(95%CI:2.6%~7.4%)。检测到33种NOTCH3基因突变,包括31种错义突变和2种剪接突变。18例患者存在导致半胱氨酸数量改变的错义突变,总发生率为5.5%(95%CI:3.3%~8.5%)。CADASIL组和非CADASIL组患者年龄、性别、痴呆家族史、血管危险因素、颅内外大动脉狭窄、梗死灶数量、前颞极白质病变、精神障碍、偏头痛比较差异均无统计学意义(P>0.05),脑微出血、脑白质变性、认知障碍差异均有统计学意义(P<0.05)。结论腔隙性梗死患者NOTCH3基因致病性突变率与既往研究结果大致一致,但c.1630C>T突变发生率高于既往估计,在发现有更多的新突变位点前,选择性对突变“热点区域”检测更经济、高效。合并脑微出血、脑白质变性、认知障碍的腔隙性梗死患者的NOTCH3基因筛查更有意义。

STAT3-Dependent Effects of Polymeric Immunoglobulin Receptor in Regulating Interleukin-17 Signaling and Preventing Autoimmune Hepatitis

One-third of patients with autoimmune hepatitis(AIH)have cirrhosis at the time of diagnosis.The relevance of these variables,although unknown,is believed to be critical in AIH because of suspected interactions between the gut microbiome and genetic factors.Dysbiosis of the gut flora and elevated polymeric immunoglobulin receptor(pIgR)levels have been observed in both patients and mouse models.Moreover,there is a direct relationship between pIgR expression and transaminase levels in patients with AIH.In this study,we aimed to explore how pIgR influences the secretion of regenerating islet-derived 3 beta(Reg3b)and the flora composition in AIH using in vivo experiments involving patients with AIH and a concanavalin A-induced mouse model of AIH.Reg3b expression was reduced in pIgR gene(Pigr)-knockout mice compared to that in wild-type mice,leading to increased microbiota disruption.Conversely,exogenous pIgR supplementation increased Reg3b expression and maintained microbiota homeostasis.RNA sequencing revealed the participation of the interleukin(IL)-17 signaling pathway in the regulation of Reg3b through pIgR.Furthermore,the introduction of external pIgR could not restore the imbalance in gut microbiota in AIH,and the decrease in Reg3b expression was not apparent following the inhibition of signal transducer and activator of transcription 3(STAT3).In this study,pIgR facilitated the upregulation of Reg3b via the STAT3 pathway,which plays a crucial role in preserving the balance of the intestinal microbiota in AIH.Through this research,we discovered new molecular targets that can be used for the diagnosis and treatment of AIH.

BIRC3 induces the phosphoinositide 3-kinase-Akt pathway activation to promote trastuzumab resistance in human epidermal growth factor receptor 2-positive gastric cancer

BACKGROUND Trastuzumab-targeted therapy is currently the standard of care for advanced human epidermal growth factor receptor 2(HER2)-positive gastric cancer.However,the emergence of resistance to trastuzumab poses significant challenges.AIM To identify the key genes associated with trastuzumab resistance.These results provide a basis for the development of interventions to address drug resistance and improve patient outcomes.METHODS High-throughput sequencing and bioinformatics were used to identify the differentially expressed pivotal gene BIRC3 and delineate its potential function and pathway regulation.Tumor samples were collected from patients with HER2-positive gastric cancer to evaluate the correlation between BIRC3 expression and trastuzumab resistance.We established gastric cancer cell lines with both highly expressed and suppressed levels of BIRC3,followed by comprehensive in vitro and in vivo experiments to confirm the involvement of BIRC3 in trastuzumab resistance and to elucidate its underlying mechanisms.RESULTS In patients with HER2-positive gastric cancer,there is a significant correlation between elevated BIRC3 expression in tumor tissues and higher T stage,tumor node metastasis stage,as well as poor overall survival and progressionfree survival.BIRC3 is highly expressed in trastuzumab-resistant gastric cancer cell lines,where it inhibits tumor cell apoptosis and enhances trastuzumab resistance by promoting the phosphorylation and activation of the phosphoinositide 3-kinase-Akt(PI3K-AKT)pathway in HER2-positive gastric cancer cells,both in vivo and in vitro.CONCLUSION This study revealed a robust association between high BIRC3 expression and an unfavorable prognosis in patients with HER2-positive gastric cancer.Thus,the high expression of BIRC3 stimulated PI3K-AKT phosphorylation and activation,stimulating the proliferation of HER2-positive tumor cells and suppressing apoptosis,ultimately leading to trastuzumab resistance.

Diabetic cardiomyopathy:Importance of direct evidence to support the roles of NOD-like receptor protein 3 inflammasome and pyroptosis

Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,most studies have focused mainly on whether diabetes increases the NLRP3 inflammasome and associated pyroptosis in the heart of type 1 or type 2 diabetic rodent models,and whether various medications and natural products prevent the development of DCM,associated with decreased levels of cardiac NLRP3 inflammasome and pyroptosis.The direct link of NLRP3 inflammasome and associated pyroptosis to the pathogenesis of DCM remains unclear based on the limited evidence derived from the available studies,with the approaches of NLRP3 gene silencing or pharmaceutical application of NLRP3 specific inhibitors.We thus emphasize the requirement for more systematic studies that are designed to provide direct evidence to support the link,given that several studies have provided both direct and indirect evidence under specific conditions.This editorial emphasizes that the current investigation should be circumspect in its conclusion,i.e.,not overemphasizing its role in the pathogenesis of DCM with the fact of only significantly increased expression or activation of NLRP3 inflammasome and pyroptosis in the heart of diabetic rodent models.Only clear-cut evidence-based causative roles of NLRP3 inflammasome and pyroptosis in the pathogenesis of DCM can help to develop effective and safe medications for the clinical management of DCM,targeting these biomarkers.

一例新发NOTCH3基因突变的伴有皮质下梗死和白质脑病的常染色体显性脑动脉病患者的临床表型和基因分析

伴有皮质下梗死和白质脑病的常染色体显性脑动脉病(cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy,CADASIL)是一种由NOTCH3突变引起的单基因遗传性脑小血管疾病,目前被认为是成人缺血性脑卒中和痴呆的最常见遗传原因。NOTCH3位于染色体19p13.1-13.2上,95%突变为杂合错义突变[1]。CADASIL主要临床特征包括先兆的偏头痛、反复缺血性脑卒中、短暂性脑缺血发作、进行性白质变性、记忆力减退、痴呆以及多种精神症状[2]。

皮质下梗死伴白质脑病的常染色体显性遗传性脑动脉病两家系NOTCH3基因突变分析

皮质下梗死伴白质脑病的常染色体显性遗传性脑动脉病(cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy,CADASIL)是一种较少见的、以反复脑卒中发作与血管性痴呆为特征的遗传性脑小血管疾病[1]。该病多于中年起病,早期可有先兆性偏头痛、短暂性脑缺血发作等症状,随着年龄增长逐渐出现反复脑卒中发作,最终导致精神异常和血管性痴呆;头颅MRI特征包括皮层下腔隙性梗死灶、颞极、侧脑室周围和皮层下白质病变[2]。CADASIL致病基因NOTCH3位于染色体19p13.2-13.1,大部分变异为错义突变,也有少部分为缺失、插入和剪切[3]。随着分子检测技术的进步和普及,越来越多的CADASIL家系经基因检测确诊[4-5]。本文分析了两个CADASIL家系NOTCH3基因突变情况,并对其临床特点进行了总结,报道如下。

肺癌中TSG101蛋白与Notch3受体表达的相关性

背景与目的TSG101蛋白是内吞分选通路的重要因子,该通路的异常会减少果蝇中Notch受体的降解,导致此受体信号对细胞分化、发育调节的紊乱。在哺乳动物中,Notch受体家族中的3亚型与肺组织发育及肺癌发生密切相关。本实验的目的是初步探讨在人类肺癌中,TSG101蛋白与Notch3受体之间的表达是否也存在关联。方法分别采用Western blot和免疫组化SP法检测TSG101蛋白与Notch3受体在肺癌组织、细胞系中的表达,并应用抗体封闭TSG101蛋白功能后观察Notch3受体的表达变化。结果TSG101蛋白在肺癌中比正常肺组织表达减少,而Notch3受体在肺癌中比正常肺组织表达则增多,此变化均与肺癌的分化及淋巴结转移相关。当用抗体封闭肺癌细胞系中TSG101蛋白功能时,Notch3受体检测水平呈上升变化。结论肺癌中TSG101蛋白表达下调,与Notch3受体检测水平上升有相关性。

Inhibition of 5-HT_3 Receptors-activated Currents by Cannabinoids in Rat Trigeminal Ganglion Neurons

This study investigated the modulatory effect of synthetic cannabinoids WIN55,212-2 on 5-HT3 receptor-activated currents (I5-HT3) in cultured rat trigeminal ganglion (TG) neurons using whole-cell patch clamp technique. The results showed that: (1) The majority of examined neurons (78.70%) were sensitive to 5-HT (3–300 μmol/L). 5-HT induced inward currents in a concentration-dependent manner and the currents were blocked by ICS 205-930 (1 μmol/L), a selective antagonist of the 5-HT3 receptor; (2) Pre-application of WIN55,212-2 (0.01–1 μmol/L) significantly inhibited I5-HT3 reversibly in concentration-dependent and voltage-independent manners. The concentra-tion-response curve of 5-HT3 receptor was shifted downward by WIN55,212-2 without any change of the threshold value. The EC50 values of two curves were very close (17.5±4.5) mmol/L vs. (15.2±4.5) mmol/L and WIN55,212-2 decreased the maximal amplitude of I5-HT3 by (48.65±4.15)%; (3) Neither AM281, a selective CB1 receptor antagonist, nor AM630, a selective CB2 receptor antagonist reversed the inhibition of I5-HT3 by WIN55,212-2; (4) When WIN55,212-2 was given from 15 to 120 s before 5-HT application, inhibitory effect was gradually increased and the maximal inhibition took place at 90 s, and the inhibition remained at the same level after 90 s. We are led to concluded that-WIN55,212-2 inhibited I5-HT3 significantly and neither CB1 receptor antagonist nor CB2 receptor antagonist could reverse the inhibition of I5-HT3 by WIN55,212-2. Moreover, WIN55,212-2 is not an open channel blocker (OCB) of 5-HT3 receptor. WIN55,212-2 significantly inhibited 5-HT-activated currents in a non-competitive manner. The inhibition of I5-HT3 by WIN55,212-2 is probably new one of peripheral analgesic mechanisms of WIN55,212-2, but the mechanism by which WIN55,212-2 inhibits I5-HT3 warrants further investigation.

Notch2、Notch3、Jagged2和Hes3在当归补血汤协同肌源性干细胞移植鼠骨髓中的表达和作用

目的探讨Notch2、Notch3、Jagged2和Hes3在当归补血汤(Danggui Buxue Decoction,DBD)协同肌源性干细胞(Muscle-derived stem cells,MDSCs)移植小鼠骨髓中的表达和作用。方法将雌性昆明小鼠随机分为6组:照射模型组、骨髓移植组、MDSCs移植组和当归补血汤不同剂量(1、3和5倍)预处理+MDSCs移植组。各组分别给予生理盐水和不同剂量DBD灌胃1周后,经8Gy^(137)Cs-γ射线照射,尾静脉分别移植骨髓细胞和MDSCs。用Real-time PCR分别检测3周和8周末时小鼠骨髓中Notch2、Notch3、Jagged2和Hes3 mRNA表达量。结果 3周末,骨髓移植组和MDSCs移植组Notch2mRNA上调,给药组下调,组间比较无统计学差异(P>0.05)。Notch3 mRNA均下调(P<0.05),且给药2组下调幅度最大;8周末,除MDSCs移植组外,其余各组Notch2 mRNA均下调(P<0.05),且以给药2组下调幅度最大。各干预组Notch3 mRNA均下调(P<0.05),其中骨髓移植组下调幅度最小;Jagged2和Hes3 mRNA均未见表达。结论 Jagged2配体和Hes3靶基因既不参与造血干细胞向造血前体细胞分化过程,也不参与MDSCs向造血干细胞分化、增殖过程。而Notch2受体和Notch3受体在这些过程中扮演不同的角色。在造血重建后期,5倍当归补血汤可通过上调Notch2和Notch3表达来促进淋巴细胞的发育。

siRNA下调Notch3表达对非小细胞肺癌骨转移能力的影响

目的:研究Notch3在肺癌骨转移中的作用。方法:通过RT-PCR方法检测Notch3在NSCLC及其伴骨转移患者的肺癌组织中的表达情况;利用慢病毒包装Notch3 siRNA载体抑制Notch3的表达后,通过体外迁移和侵袭试验观察Notch3 siRNA对肺癌细胞的迁移和侵袭能力的影响;通过体外ELISA法检测骨转移相关分子pTHrP和IL-6的表达,研究阻断Notch3的表达后是否能够逆转TGF-β诱导的肺癌骨转移。结果:Notch3在NSCLC发生骨转移的患者肺癌组织中过表达。体外试验发现,通过慢病毒包装Notch3 siRNA载体抑制Notch3的表达后,NSCLC细胞的转移能力下降,且Notch3-si可明显抑制NSCLC细胞中由TGF-β介导的pTHrP和IL-6的表达。结论:Notch3高表达在促进NSCLC骨转移中发挥重要作用。

Notch3在卵巢上皮性癌中的表达及临床意义

目的探讨卵巢上皮性癌组织中Notch3的表达与卵巢癌血管生成以及临床病理特征的关系。方法应用免疫组织化学SP法检测91例卵巢肿瘤组织中Notch3的表达和CD34标记的微血管密度（MVD）,结合临床病理特征进行综合分析。结果在恶性和交界性卵巢肿瘤组织中Notch3阳性率及MVD计数均显著高于正常卵巢和良性卵巢肿瘤（均P〈0.01）。Notch3在FIGOⅢ-Ⅳ期,低分化和有腹水的卵巢癌患者中的阳性率高于FIGOⅠ-Ⅱ期,高、中分化和无腹水者（均P〈0.05）,MVD计数与患者组织学分级和有无腹水相关（均P〈0.05）,Notch3在卵巢癌组织中的表达与MVD计数之间存在相关性（rs=0.531,P〈0.01）。结论 Notch3可能参与卵巢癌的血管生成,促进卵巢癌的发生发展过程。

Notch2和Notch3对滋养细胞株迁移与侵袭功能的影响

目的探讨Notch2和Notch3对BeWo及JAR人胎盘绒毛癌滋养细胞株迁移和侵袭能力的影响。方法通过对BeWo及JAR细胞Notch2和Notch3基因的过表达及干扰,观察两种滋养细胞株迁移、侵袭生物学功能的变化。结果在BeWo细胞中对Notch2干扰后细胞迁移能力在48 h明显减弱,而72 h的迁移能力及Notch2干扰后的细胞侵袭能力无显著变化;Notch3过表达后细胞迁移能力在48 h显著增强,而72 h则显著减弱;Notch3过表达后细胞侵袭能力在48 h、72 h均显著加强。在JAR细胞中对Notch3干扰后细胞迁移能力在48 h、72 h均明显减弱;而Notch3干扰后细胞的侵袭能力及Notch2过表达后细胞迁移能力、侵袭能力均无明显变化。结论 Notch2、Notch3影响BeWo及JAR细胞迁移、侵袭功能。

野百合碱诱导大鼠肺动脉高压时对肺血管壁Jagged2/Notch3信号分子表达的影响

目的:观察野百合碱诱导大鼠肺动脉高压时肺血管壁Jagged2/Notch3信号分子表达的变化,探讨Jagged2/Notch3信号在肺动脉高压发生中的作用和意义。方法:45只SD大鼠随机分为正常对照组(C组)、溶剂对照组(S组)和野百合碱模型组(M组),每组15只,通过一次性腹腔注射野百合碱50 mg/kg建立肺动脉高压模型,溶剂对照组注射相同剂量的溶媒。4周时通过HE染色观察肺血管重构,通过右心导管测定平均肺动脉压(m PAP)和右心室收缩压(RVSP)。采用免疫组化和实时荧光定量PCR等方法检测肺血管壁Jagged2/Notch3/Hes5蛋白和mRNA表达的变化。结果:与正常对照和溶剂对照组相比,4周时野百合碱模型组的血管壁显著增厚,中膜厚度百分比增加(P<0.01);此外野百合碱模型组的m PAP和RVSP显著高于正常对照和溶剂对照组(P<0.01);免疫组化和实时荧光定量PCR结果提示Jagged2主要表达于肺小动脉内膜,Notch3、Hes5主要表达于肺小动脉中层平滑肌,与溶剂对照组以及正常对照组相比,野百合碱模型组肺小动脉的Jagged2、Notch3和Hes5表达显著增高。结论:Jagged2/Notch3信号分子的激活可能在野百合碱诱导肺动脉高压发生中起重要作用。