YKL-40 和NF-κ B 在子宫内膜异位症中的表达及相关性研究

目的:通过检测人类软骨蛋白39(YKL-40)和核转录因子KappaB(NF-κB)在子宫内膜异位症组织中的表达,探讨二者与子宫内膜异位症的关系及二者的相关性。方法:用免疫组化二步法检测40例子宫内膜异位症(EMS)患者的异位内膜和在位内膜及40例子宫肌瘤患者子宫内膜(对照组)YKL-40和NF-κB的表达,并对检测结果进行统计学分析。结果:YKL-40、NF-κB在子宫内膜异位症患者的异位和在位内膜及对照组内膜中的表达率分别为67.5%、62.5%、35%及88.5%、57.5%、32.5%,差异有统计学意义(P<0.01);在不同月经周期YKL-40表达无统计学意义;在位内膜和正常内膜NF-κB的表达分泌期高于增殖期,差异有统计学意义(P<0.05);YKL-40和NF-κB在三种内膜中的表达具有正相关性,相关系数分别是0.305,0.267和0.457(P<0.01)。结论:YKL-40和NF-κB在EMS发生发展中起重要的作用。     

PTEN蛋白的异常表达与卵巢子宫内膜异位症恶变的关系

目的:探讨PTEN蛋白表达与与卵巢子宫内膜异位症(内异症)恶变的关系.方法:应用免疫组化方法检测21例子宫内膜异位症相关卵巢癌(内异症相关卵巢癌),10例不典型卵巢子宫内膜异位症(不典型内异症)及23例内异位症中PTEN蛋白表达.结果:PTEN在内异症和不典型内异症组阳性表达率分别为82.61%和60.00%,内异症相关卵巢癌组中PTEN蛋白阳性表达率为42.86%.内异症相关卵巢癌组PTEN蛋白表达与内异症组相比差异有显著性(p＜0.05). PTEN蛋白表达与内异症相关卵巢癌患者的年龄、病理类型无相关性(p＞0.05),与分化程度、临床分期相关(p＜0.05),在内异症相关卵巢癌中,临床分期早,分化程度高的PTEN表达显著高于分期晚,分化差的.结论:PTEN蛋白表达缺失可能参与了卵巢子宫内膜异位症恶变,有可能成为一个很有潜力的诊断指标及新的治疗靶点而应用于临床.     

VEGF和IGF-I在子宫内膜异位症患者血清中的表达及临床意义

目的：探究血管内皮生长因子（VEGF）和胰岛素样生长因子-I（IGF-I）在子宫内膜异位症患者血清中的表达及临床意义,为 子宫内膜异位症的治疗提供参考。方法：选取我院2015 年1 月至2016 年1 月收治的子宫内膜异位症患者50 例为实验组,另选 体检中心健康妇女50 例为对照组。实验组患者根据疾病不同分期分为I、II期（n=24）和III、IV 期（n=26）。通过酶联免疫吸附法 （ELISA）检测两组对象血清中VEGF和IGF-I的水平,采用Pearson相关分析法分析实验组患者血清中VEGF 和IGF-I 表达的相 关性。结果：实验组患者血清中VEGF和IGF-I的水平均明显高于对照组,差异有统计学意义（P<0.05）;实验组III、IV期患者血清 中VEGF和IGF-I的水平明显高于I、II期患者,差异有统计学意义（P<0.05）;Pearson 相关性分析显示实验组患者血清中VEGF 和IGF-I的水平变化呈正相关关系（r=0.507,P<0.05）。结论：子宫内膜异位症患者血清中VEGF和IGF-I的水平高于正常水平,并 随着病情的加重而不断升高,且二者呈正相关关系,可协同作用加快病情发展。     

MMP-2、MMP-9及VEGF在子宫内膜异位症中的表达及临床意义

目的:研究基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、血管内皮生长因子(VEGF)表达在子宫内膜异位症发生发展中的作用.方法:应用免疫组化二步法检测EMs患者异位内膜36例、在位内膜36例及正常内膜中的MMP-2、MMP-9、VEGF的表达情况.结果:MMP-2、MMP-9、VEGF在异位内膜组中阳性表达率分别为94.4%、91.7%和91.7%,显著高于在位内膜组、正常内膜组(P＜0.05);而在位内膜组和正常内膜组差异无统计学意义(P＞0.05).结论:检测MMP-2、MMP-9、VEGF的表达可用于判断子宫内膜异位症的侵袭转移和评估预后.     

P物质在子宫内膜异位症中的表达及其意义

目的:探讨P物质(substance P,SP)在子宫内膜异位症(endometriosis,EM)中的表达及其意义。方法:采用免疫组织化学法检测2012年10月至2013年4月在哈尔滨医科大学附属第一医院经腹腔镜及病理证实的EM患者异位子宫内膜组织20例,与其配对的在位子宫内膜组织10例以及因非EM(子宫肌瘤)行腹腔镜下子宫全切术或肌瘤核除患者的正常子宫内膜组织20例中SP的表达情况,并分析和比较术中盆腔粘连发生情况。结果:SP在EM患者的异位子宫内膜、在位子宫内膜及非EM患者的正常子宫内膜的阳性表达率分别为75%、80%、20%,异位子宫内膜和在位子宫内膜比较无显著性差别(P=1.0),但均高于非EM患者的正常子宫内膜,差异均有统计学意义(P=0.002,P=0.004);EM组盆腔粘连的阳性率高于非EM组,EM患者异位子宫内膜中SP阳性组盆腔粘连阳性率高于SP阴性组,差异均有统计学意义(P=0.001,P=0.032),非EM患者正常子宫内膜中SP阳性组盆腔粘连阳性率与SP阴性组相比,差异不具有统计学意义(P=0.061)。结论:SP在EM的异位子宫内膜和在位子宫内膜中的表达上调,并与EM合并盆腔粘连有关,其具体机制尚有待进一步的研究。     

改良大鼠子宫内膜异位症模型的建立及微血管密度观察

目的对皮下筋膜层与腹壁肌层之间移植自体子宫内膜制作的子宫内膜异位症（endometriosis,EMs）模型进行评估。方法取10只雌性未交配性成熟大鼠,术前雌激素诱导,手术开腹取右侧子宫,将自体子宫内膜种植于双侧皮下筋膜层与腹壁肌层之间,术后第29天取出异位组织,进行组织形态学观察,免疫组织化学染色观察微血管密度（Microvessel density,MVD）。结果异位内膜在腹壁内生长,呈隆起囊状小包块,内有黏液,具有正常子宫内膜基本组织结构。异位内膜中微血管密度较在位内膜和正常子宫内膜高。结论此手术方法建立的大鼠子宫内膜异位症模型异位内膜病理改变与EMs患者类似,可以作为子宫内膜异位研究的动物模型。     

上皮间质转化标记物及Snail在子宫内膜异位症中的表达

目的:研究上皮间质转化标志物(E-cadherin、β-catenin、vimentin)和Snail在子宫内膜异位症(endometriosis,EMs)中的表达。方法:选取40例EMs患者(实验组)异位内膜及在位内膜,同时获取20例非EMs患者(对照组)的正常子宫内膜,采用免疫组化法研究Snail、EMT上皮标志物(E-cadherin、β-catenin)、间质标志物(vimentin)在各内膜组织中的表达,并比较其表达水平。结果:EMs患者异位内膜和在位内膜的EMT上皮标志物E-cadherin、β-catenin表达均显著低于正常内膜的表达(P0.05);EMs患者异位内膜和在位内膜的EMT间质标志物vimentin表达均显著高于正常内膜的表达(P0.05);EMs患者异位内膜和在位内膜中Snail表达显著高于正常内膜的表达(P0.05)。结论:在子宫内膜异位症(EMs)中,Snail、vimentin表达上调,E-cadherin、β-catenin表达下调可能与子宫内膜异位症(EMs)的发生、发展及浸润转移有关。     

子宫内膜异位症患者凋亡相关基因TGF-β mRNA表达的研究

目的:探计TGF-β在子宫内膜异位症发病及发展中的分子生物学机制及与凋亡的相关性.方法:选取子宫内膜异位症患者12例为研究对象(EMS组),正常妇女正常子宫内膜12例(对照组),应用半定量逆转录-聚合酶链反应(RT-PCR)方法,测定TGF-β mRNA表达水平;运用western-blotting印迹法检测检测TGF-β蛋白的表达.同时用TUNEL染色对标本进行凋亡检测.结果:异位子宫内膜的凋亡率明显低于正常子宫内膜,且EMS组患者异位内膜的TGF-β mKNA和蛋白的表达水平上调.结论:患者子宫内膜中凋亡率的改变,在子宫内膜异位症的发生发展中起重要作用.异位的子宫内膜可能是由于TGF-β的表达上调增加了抗凋亡能力.     

EFI评分、血清MMIF、YKL-40与子宫内膜异位症患者卵巢功能、临床分期的关系

摘要 目的：探讨生育指数（EFI）评分、血清巨噬细胞移动抑制因子（MMIF）、甲壳质酶蛋白 40（YKL-40）与子宫内膜异位症患者卵巢功能、临床分期的关系。方法：选取我院2020年6月到2023年6月收治的80例子宫内膜异位症患者,分为观察组,另选取同期来我院体检的80名健康女性志愿者作为对照组。对比两组受检者EFI评分、血清MMIF、YKL-40及卵巢功能指标表达水平。应用美国生殖医学协会子宫内膜异位症分期标准(r-AFS)对80例子宫内膜异位症患者进行分期,其中Ⅰ期15例,Ⅱ期23例,Ⅲ期25例,Ⅳ期17例,对比不同r-AFS分期患者EFI评分、血清MMIF、YKL-40表达水平。并采用Spearman相关法分析法分析EFI评分、血清MMIF、YKL-40与卵巢功能指标的相关性。结果：观察组患者EFI评分低于对照组,血清MMIF、YKL-40表达水平高于对照组（P＜0.05）;观察组抗苗勒氏管激素（AMH）、黄体生成素（LH）、卵泡刺激素（FSH）表达水平低于对照组,雌二醇（Estradiol,E₂）水平高于对照组（P＜0.05）;不同分期子宫内膜异位症患者EFI评分由高到低分别为Ⅰ期、Ⅱ期、Ⅲ期、Ⅳ期,MMIF和YKL-40水平由高到低分别为Ⅳ期、Ⅲ期、Ⅱ期、Ⅰ期,不同分期EFI评分、血清MMIF、YKL-40表达水平对比差异显著（P＜0.05）;Spearman相关法分析结果表明：EFI评分与AMH、LH、FSH呈正相关,与E2呈负相关（P＜0.05）,MMIF和YKL-40与AMH、LH、FSH呈负相关,与E₂呈正相关（P＜0.05）。结论：EFI评分、血清MMIF、YKL-40水平与子宫内膜异位症患者的卵巢具有明显相关性,且不同子宫内膜异位症患者EFI评分、血清MMIF、YKL-40水平具有显著差异,临床可考虑应用EFI评分、血清MMIF、YKL-40水平来辅助判断子宫内膜异位症患者的卵巢功能与疾病严重程度。     

基质金属蛋白酶-2 在子宫内膜异位症中的表达及应用价值

目的:探讨粘附分子CD44拼构变异体6(CD44v6)和基质金属蛋白酶-2(MMP-2)在子宫内膜异位症(EMs)组织中的表达及相关性。方法:选取20例异位内膜组织标本、20例在位内膜组织标本及20例正常子宫内膜标本,用病理常规免疫组织化学方法检测MMP-2和CD44v6的表达,并分析其相关性。结果:CD44v6在异位内膜组的表达明显高于在位内膜组和对照组,且对照组明显高于在位内膜组,差异具有统计学意义(P0.05);CD44v6在在位内膜组和对照组中分泌期的表达明显高于同组增生期,差异具有统计学意义(P0.05)。MMP-2在异位内膜组和在位内膜组的表达明显高于对照组,差异具有统计学意义(P0.01);MMP-2在各组增生期和分泌期表达不规律。异位内膜组中,CD44v6和MMP-2在Ⅲ-Ⅳ期的表达明显高于Ⅰ-Ⅱ期,差异具有统计学意义(P0.01)。Spearman相关性分析结果显示:EMs组织中MMP-2和CD44v6之间呈现正相关性(r=0.724,P0.05);EMs不同分期组织中MMP-2和CD44v6之间亦呈现正相关性(r=0.623,P0.05)。结论:MMP-2和CD44v6在EMs异位内膜中高表达,且有正协同作用,二者可能与EMs的发生发展有关。     

YKL-40在脑胶质母细胞瘤微环境中的作用

胶质母细胞瘤是大脑及其他中枢神经系统最常见的恶性肿瘤,其复杂的肿瘤微环境是胶质母细胞瘤临床治疗的主要挑战,也是胶质母细胞瘤患者复发率高、生存率低的主要原因。YKL-40,这一分泌性蛋白质与多种类型的癌症预后不良相关,且在高级别胶质瘤尤其是胶质母细胞瘤患者中血清水平与肿瘤组织表达水平显著升高,而在低级别胶质瘤中并未发现这一特征。这提示,YKL-40与胶质瘤分级及胶质母细胞瘤恶性发展过程密切相关。针对YKL-40的抗体治疗也被证明能够与电离辐射协同抑制胶质母细胞瘤血管生成及恶性发展。基于YKL-40的临床价值,本文将从肿瘤微环境的角度,归纳总结YKL-40在恶性肿瘤中的相关研究成果,并讨论其在胶质母细胞瘤发生发展中的相关作用及临床应用前景。     

老年2型糖尿病患者血清BGP、ghrelin、YKL-40水平与胰岛素抵抗及认知功能损害的关系研究

摘要 目的：研究老年2型糖尿病（T2DM）患者血清骨钙素（BGP）、生长激素释放多肽（ghrelin）及人软骨糖蛋白-39（YKL-40）水平与胰岛素抵抗及认知功能损害的关系。方法：选择2018年1月～2019年12月我院收治的老年T2DM患者100例记作病变组,选择同期于我院进行体检的老年健康志愿者100例作为对照组。比较两组血清BGP、ghrelin及YKL-40水平,血糖、血脂、胰岛素抵抗相关指标及简易智力状态检查量表（MMSE）评分情况,并分析各指标的相关性。结果：病变组血清BGP、ghrelin水平低于对照组,而YKL-40水平高于对照组（P＜0.05）。病变组空腹血糖（FPG）、糖化血红蛋白（HbA1c）水平及胰岛素抵抗指数（HOMA-IR）均高于对照组,而空腹胰岛素（FINS）水平低于对照组（P＜0.05）。病变组除语言、延迟记忆评分与对照组对比差异无统计学意义（P＞0.05）,其他各项MMSE评分均低于对照组（P＜0.05）。经Pearson相关性分析可得：老年T2DM患者血清BGP、ghrelin水平和FPG、HbA1c、HOMA-IR均呈负相关,与FINS及MMSE评分均呈正相关（P＜0.05）;而血清YKL-40水平和FPG、HbA1c、HOMA-IR均呈正相关,与FINS及MMSE评分均呈负相关（P＜0.05）。结论：老年T2DM患者的血清BGP、ghrelin存在明显低表达,而血清YKL-40水平呈明显高表达,且上述血清学指标水平均与胰岛素抵抗和认知功能损害密切相关。     

Association between serum YKL-40 level and dysglycemia in cystic fibrosis

BackgroundYKL-40, a chitinase-like protein, is a biomarker for type 1 and type 2 diabetes prognosis. We hypothesized that YKL-40 protein levels are elevated in CF patients with dysglycemia.MethodsSeventeen healthy control subjects and 66 CF patients were prospectively recruited and subjected to an oral glucose tolerance test. In all participants, fasting serum YKL-40 was compared between control and CF patients and between normal glucose-tolerant patients (NG-CF) and CF patients with dysglycemia (DG-CF). A Botnia clamp procedure was performed on a subset of patients for each group to determine the impact of acute increases of either glucose or insulin on YKL-40 concentration.ResultsCF patients had higher serum YKL-40 values than the controls (113 [49;288] vs. 38 [30;50] ng/ml, p < 0.001). YKL-40 concentrations in CF patients were mainly increased in the DG-CF group, who had significantly higher values: 213 [93;383] vs. 67 [27;97] ng/ml in the NG-CF group, p < 0.001). No significant modulation of YKL-40 concentration was observed in serum of CF (NG or DG-CF) or non-CF patients, after acute exposure to glucose or insulin.ConclusionsHigher serum YKL-40 levels in CF patients are significantly associated with dysglycemia. The increase in YKL-40 is potentially associated with an inflammatory response resulting from chronic glucose intolerance or CF disease evolution.     

The chitinase-like protein YKL-40 increases mucin5AC production in human bronchial epithelial cells

Mucus overproduction is an important feature in patients with chronic inflammatory airway diseases. However, the regulatory mechanisms that mediate excessive mucin production remain elusive. Recently, the level of YKL-40, a chitinase-like protein, has been found to be significantly increased in chronic inflammatory airway diseases and has been shown to be associated with the severity of these diseases. In this study, we sought to explore the effect of YKL-40 on mucin5AC (MUC5AC) production in chronic inflammatory airway diseases and the potential signaling pathways involved in this process. We found that elevated YKL-40 levels increased the mRNA and protein expression of MUC5AC in a dose- and time-dependent manner, in association with the phosphorylation of extracellular signal-regulated kinase (ERK) and nuclear factor κB (NF-κB), reflecting their activation. These responses were significantly suppressed by the knockdown of protease-activating receptor 2 (PAR2) with specific small interfering RNA or the inhibitors of ERK and NF-κB. YKL-40-induced MUC5AC overproduction was also effectively attenuated by the inhibitor of focal adhesion kinase (FAK). Taken together, these results imply that YKL-40 can stimulate excessive MUC5AC production through PAR2- and FAK-mediated mechanisms.     

The role of YKL-40 in the pathogenesis of autoimmune diseases: a comprehensive review

YKL-40, a chitinase-3-like protein 1 (CHI3L1) or human cartilage glycoprotein 39 (HC gp-39), is expressed and secreted by various cell-types including macrophages, chondrocytes, fibroblast-like synovial cells and vascular smooth muscle cells. Its biological function is not well elucidated, but it is speculated to have some connection with inflammatory reactions and autoimmune diseases. Although having important biological roles in autoimmunity, there were only attempts to elucidate relationships of YKL-40 with a single or couple of diseases in the literature. Therefore, in order to analyze the relationship between YKL-40 and the overall diseases, we reviewed 51 articles that discussed the association of YKL-40 with rheumatoid arthritis, psoriasis, systemic lupus erythematosus, Behçet disease and inflammatory bowel disease. Several studies showed that YKL-40 could be assumed as a marker for disease diagnosis, prognosis, disease activity and severity. It is also shown to be involved in response to disease treatment. However, other studies showed controversial results particularly in the case of Behçet disease activity. Therefore, further studies are needed to elucidate the exact role of YKL-40 in autoimmunity and to investigate its potential in therapeutics.     

YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers

The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3β, PDX1, CD34, p63, nestin, PAX6) markers. Double-labeling showed YKL-40 expression in OCT4-positive hESCs, PAX6-positive neuroectodermal cells, and HNF-3β-positive endodermal cells. The differentiating progeny showed strong YKL-40 expression. Abrupt transition between YKL-40 and OCT4-positive hESCs and YKL-40-positive ecto- and neuroectodermal lineages was observed within the same epithelial-like layer. YKL-40-positive cells within deeper layers lacked contact with OCT4-positive cells. YKL-40 may be important in initial cell differentiation from hESCs toward ectoderm and neuroectoderm, with retained epithelial morphology, whereas later differentiation into endoderm and mesoderm involves a transition into the deeper layers of the colony.     

Knockdown of YKL-40 inhibits angiogenesis through regulation of VEGF/VEGFR2 and ERK1/2 signaling in endometrial cancer

Studies have demonstrated that small interfering RNA (siRNA) targeting YKL-40 (siYKL-40) inhibits the proliferation, migration, invasion, and induces antiapoptotic abilities of endometrial cancer (EC) HEC-1A cells. However, its effect on angiogenesis is unclear. The present study aimed to investigate the role of YKL-40 in endometrial cancer and the related molecular mechanisms. YKL-40 was knocked down by transfection with siYKL-40 and the effects on angiogenesis, cell viability, and signaling pathways were investigated. The results showed that siYKL-40 inhibited VEGFA levels and tube formation in endothelial cells. Additionally, inhibition of YKL-40 decreased the expression levels of vascular endothelial growth factor (VEGF), phosphorylated vascular endothelial growth factor receptor 2 (pVEGFR2), and phosphorylated extracellular signal-regulated kinases 1 and 2 (pERK1/2). Furthermore, a nude mice xenograft model of EC showed that siYKL-40 inhibited tumor growth. Inhibition of YKL-40 led to suppression of angiogenesis and reduction of microvessel density through VEGF/VEGFR2 and ERK1/2 signaling in endometrial cancer cells. Taken together, this study demonstrated novel molecular mechanisms for role of YKL-40 in EC.     

分泌蛋白YKL-40增强肿瘤细胞的上皮间质样转化

目的:分泌糖蛋白YKL-40在多种晚期肿瘤病人的血液中显著升高,提示YKL-40蛋白的血浓度是肿瘤恶变的生物标志物。本课题研究YKL-40重组蛋白和过表达YKL-40肿瘤细胞对肿瘤细胞的上皮间质样转化的作用。方法:构建YKL-40过表达的纤维状乳腺癌细胞系MDA-MB-231和非纤维状结肠癌细胞系HCT-116,观察细胞形态学变化,收集细胞和细胞培养液用于Western Blot(WB)检测YKL-40和上皮间质转化标记蛋白Vimentin和N-cadherin。观察重组蛋白YKL-40对原代MDA-MB-231细胞在无血清条件下的细胞存活影响;另外,用细胞存活试剂盒检测YKL-40过表达HCT-116细胞在无血清的培养液中细胞存活情况。最后,用细胞侵袭试验检测YKL-40过表达MDA-MB-231细胞的侵袭力,并用WB和Zymography来测定细胞分泌MMP9蛋白的表达和酶活性。结果:YKL-40过表达增强MDA-MB-231细胞的形态向上皮间质样转化,并显著提高Vimentin、N-cadherin蛋白的表达,但对HCT-116细胞无法诱导上皮间质样转化。在无血清培养基培养条件下,YKL-40可以增强两种细胞的存活能力,并且YKL-40过表达的MDA-MB-231细胞增强了细胞的侵袭能力,促进了MMP9蛋白表达和蛋白活性。结论:YKL-40可以增加肿瘤细胞的存活力,增强纤维状细胞向上皮间质样转化;并且,YKL-40增加MMP9蛋白表达和活性,增强细胞侵袭力。YKL-40是间质样肿瘤细胞EMT的增强子,此发现为抑制肿瘤恶变提供新靶点。