腰椎管狭窄及肥厚黄韧带中细胞因子的表达

背景:临床上大多数研究都是针对肥厚黄韧带的形态学与组织学方面的研究,而对于黄韧带肥厚在腰椎管狭窄症中的病因学意义,目前尚缺乏统一的认识,尤其在细胞因子水平上的研究甚少。目的:通过在细胞因子水平的研究探讨腰椎黄韧带变性、肥厚的病因及其与腰椎管狭窄症的关系。方法:选择华北煤炭医学院附属骨科医院(唐山市第二医院)2008/2009腰椎管狭窄症和椎间盘突出症住院患者各25例,比较2组患者黄韧带中白细胞介素6、转化生长因子β1、肿瘤坏死因子α阳性表达程度及黄韧带中3种炎性因子阳性表达率。结果与结论:两组患者黄韧带中白细胞介素6、转化生长因子β1、肿瘤坏死因子α阳性表达程度比较,腰椎管狭窄症组明显高于腰椎间盘突出症组(P0.01)。提示白细胞介素6、转化生长因子β1、肿瘤坏死因子α在椎管狭窄症肥厚黄韧带中的表达呈相对一致性,有80%左右均明显增高。黄韧带变性、增殖、肥厚,也是无菌性炎性因子作用的结果。     

PKC信号通路与醛糖还原酶对转化生长因子β1诱导人肾系膜细胞纤连蛋白表达的影响

目的 探讨PKC信号通路与醛糖还原酶在非高糖条件下对转化生长因子(TGF)-β1诱导人肾系膜细胞(HMC)细胞外基质成分纤连蛋白表达的影响.方法 应用醛糖还原酶抑制剂、PKC信号通路抑制剂G(O)6983、转染pcDNA3-醛糖还原酶及siRNA分别作用于人系膜细胞后,再用TGF-β1刺激,观察刺激前后人系膜细胞表达纤连蛋白的情况.Western blot检测系膜细胞内纤连蛋白和醛糖还原酶的变化,即时RT-PCR鉴定转染和干扰效果.结果 系膜细胞在TGF-β1作用后,醛糖还原酶和纤连蛋白表达升高;单独使用醛糖还原酶抑制剂并不能下调纤连蛋白的表达;但预先使用醛糖还原酶抑制剂孵育后,再用TGF-β1刺激,纤连蛋白表达减少为对照组的34%(P<0.05).单独使用PKC信号通路抑制剂并不能下调纤连蛋白的表达;用PKC信号通路抑制剂后,再用TGF-β1刺激,纤连蛋白表达下降为对照组的42%(P<0.05).预先使用醛糖还原酶抑制剂、PKC抑制剂孵育细胞后,再用TGF-β1刺激,纤连蛋白表达下降;转染pcDNA3-醛糖还原酶后,系膜细胞中醛糖还原酶mRNA表达增多超过10倍,纤连蛋白表达增加了2.5倍(P<0.05),再用TGF-β1刺激,纤连蛋白表达增加了3.6倍(P<0.05);转染siRNA后,系膜细胞中醛糖还原酶mRNA表达减少为对照组的20%,纤连蛋白表达减少为对照组的6%(P<0.01),即使再用TGF-β1刺激,纤连蛋白表达仍然下降,为对照组的12%(P<0.01).结论 醛糖还原酶基因参与了系膜细胞中TGF-β1诱导纤连蛋白表达过程的调控,这一过程可能与PKC信号通路的活化有关.     

肥厚腰椎黄韧带中成纤维细胞生长因子、转化生长因子及结缔组织生长因子的表达

背景:腰椎黄韧带肥厚是临床上引起腰椎管狭窄的主要因素之一,但其分子机制仍不是非常清楚。目的:分析纤维化相关细胞因子碱性成纤维细胞生长因子、转化生长因子β1和结缔组织生长因子在腰椎黄韧带肥厚过程中的作用。方法:取临床手术所取黄韧带,对照组6例(椎管内占位且无腰椎不稳患者黄韧带)、突出组(单纯腰椎间盘突出症患者黄韧带)6例、腰椎管狭窄症组6例。采用实时定量RT-PCR的方法检测各组黄韧带中碱性成纤维细胞生长因子、转化生长因子β1、结缔组织生长因子及Ⅰ、Ⅲ、Ⅴ型胶原蛋白的mR NA含量,分析3个细胞因子在黄韧带肥厚过程中的作用。结果与结论:腰椎管狭窄症组碱性成纤维细胞生长因子mR NA表达均明显高于突出组和对照组(均P0.05);腰椎管狭窄症组转化生长因子β1 m RNA在3组中的表达明显高于对照组和突出组(均P0.01);结缔组织生长因子mR NA 3组间差异无显著性意义(P0.05)。腰椎管狭窄症组Ⅰ型胶原蛋白mR NA表达明显高于突出组和对照组(均P0.05);Ⅲ型胶原蛋白、Ⅴ型胶原蛋白mR NA表达3组之间差异无显著性意义(P0.05)。结果说明碱性成纤维细胞生长因子、转化生长因子β1在腰椎黄韧带肥厚形成过程中有重要作用,引起黄韧带肥厚的主要胶原产物为Ⅰ型胶原蛋白。     

气虚血瘀型腰椎管狭窄症患者增生肥厚与正常黄韧带之间的蛋白质表达差异

背景：从西医的病理机制来看,黄韧带增生肥厚是腰椎管狭窄症的关键致病因素,目前缺乏气虚血瘀型腰椎管狭窄症的生物学信息。目的：分析气虚血瘀型腰椎管狭窄症患者增生肥厚黄韧带与同证患者正常黄韧带之间的蛋白质表达差异。方法：选择2022年2-8月广东省第二中医院(黄埔医院)收治的气虚血瘀型腰椎管狭窄症患者6例,其中黄韧带增生肥厚者3例(实验组),黄韧带厚度正常者3例(对照组),采集所有患者黄韧带组织标本,进行4D Label free定量蛋白组学检测,筛选差异蛋白,运用GO、KEGG进行富集分析。结果与结论：(1)实验组与对照组表达差异的蛋白总数为183个,其中上调蛋白87个,下调蛋白96个;(2)表达差异蛋白的GO富集分析显示,生物进程主要集中在细胞进程、生物调节、对刺激的反应等;细胞组成则集中在细胞、细胞内、蛋白质复合物种;分子功能主要为连接、催化活性、分子功能调节剂等;(3)上调蛋白主要富集到溶酶体信号通路、类风湿性关节炎信号通路、金黄色葡萄球菌感染信号通路;下调蛋白共富集到8条信号通路,分别为p53信号通路、肾细胞癌信号通路、转化生长因子β信号通路、泛素介导的蛋白水解作用信号通路、Ca信...     

微小RNA-7基因敲减对LPS诱导的脑部炎症的影响

为研究微小RNA-7(microRNA-7, miR-7)基因敲减(knock down, KD)对LPS诱导的脑部炎症的影响并探讨其意义,课题组用LPS腹腔注射(2.5mg/kg体质量)WT小鼠建立脑部炎性损伤模型;HE染色观察小鼠脑组织病理学变化,real-time PCR探针法检测脑组织中miR-7的表达变化;进一步,用LPS腹腔注射(2.5mg/kg体质量)WT小鼠和miR-7KD小鼠;12h后,HE染色观察小鼠脑组织病理学变化;real-time PCR检测脑组织中炎性细胞因子IL-6、TNF-α和TGF-β的mRNA变化水平;ELISA检测脑组织中炎性细胞因子IL-6、TNF-α和TGF-β的变化水平;Western blotting检测信号通路Akt和p-Akt以及炎性信号通路NF-κB和p-NF-κB的表达变化。结果显示小鼠炎性损伤模型脑组织中,HE示WT小鼠在注射LPS后,脑组织中炎性细胞浸润显著增多且miR-7的表达水平显著升高(P0.01);HE结果示miR-7 KD小鼠脑组织中炎性细胞浸润较WT小鼠显著增多;real-time PCR结果示促炎细胞因子IL-6(P0.01)和TNF-α(P0.01)的表达水平均显著升高,而抑炎细胞因子TGF-β(P0.01)的表达水平显著降低;ELISA结果显示,促炎细胞因子IL-6(P0.01)和TNF-α(P0.01)的表达水平均显著升高,而抑炎细胞因子TGF-β的表达水平显著降低(P0.01); Western blotting结果示miR-7 KD模型小鼠脑组织Akt和p-Akt表达没有明显变化,NF-κB(P0.05)和p-NF-κB(P0.01)表达均明显增加。由此miR-7基因敲减显著促进了脑组织炎性损伤的发生,与NF-κB信号通路传递改变相关,本研究为后续深入探讨miR-7在脑部炎症相关疾病的发生机制中的作用提供了前期实验基础。     

血液透析中促炎性细胞因子和抗炎性细胞因子的变化及意义

促炎性细胞因子如TNF-α、IL-1β、IL-6与抗炎性细胞因子的IL-10的异常释放均与血液透析(HD)相关性炎症反应休戚相关。为了探讨促炎性细胞因子和抗炎性细胞因子在血液透析中动态变化及其意义，本研究采用随机分组的方法检测上述细胞因子在HD前后的血清浓度，以及单个淋巴细胞内上述细胞因子的表达水平。     

正五聚环蛋白3在缺血性中风发病中的作用研究进展

<正>研究表明脑缺血损伤是一种损伤级联反应,即兴奋性氨基酸毒性释放,梗死区周围去极化,程序性细胞死亡和炎性反应~([1]),其中炎性级联反应在急性缺血性卒中(acute ischemic stroke,AIS)的进展中起至关重要的作用~([2])。脑缺血可诱导多种促炎介质的表达,如细胞因子,包括肿瘤坏死因子TNF-α、白介素IL-1β、IL-10和黏附分子(ICAM)等,在中枢神经系统炎性反应的扩散和持续方面起着重要的作用~([3])。急     

没食子酸减轻香烟烟雾诱导的小鼠气道炎性反应

目的 研究没食子酸(GA)对香烟烟雾(CS)诱导的小鼠慢性阻塞性肺病(COPD)相关的肺部炎性反应的作用。方法 构建CS诱导的小鼠模型。分析支气管肺泡灌洗液中的炎性细胞的数量和促炎细胞因子的水平。相应试剂盒检测肺组织匀浆中活性氧(ROS)、谷胱甘肽(GSH)、丙二醛(MDA)和蛋白羰基的活性。Western blot分析IL-13/STAT6通路活性。结果 GA抑制SC诱导的炎性细胞浸润(中性粒细胞、淋巴细胞和巨噬细胞)和促炎细胞因子(IL-6、TNF-α和IL-1β)的水平。GA可以减轻暴露于CS的小鼠肺组织氧化还原失衡。结论 GA有效地减轻COPD模型小鼠肺部炎性反应和氧化应激。     

p53、TGF-β1及炎性细胞因子在新型酒精性肾损伤小鼠模型中的表达

鉴于以往酒精性肾损伤模型的局限性,模拟人类常见饮酒模式,将20只小鼠随机分为对照组和酒精组,采用慢性酒精喂养加单次急性酒精灌胃的方法,建造新型酒精性肾损伤小鼠模型。通过苏木精-伊红染色法(hematoxylin-eosin staining, HE)、过碘酸-雪夫染色法(periodic acid-Schiff staining,PAS)染色观察肾脏组织病理学改变,实时荧光定量PCR检测转化生长因子β1(transforming growth factor beta 1,TGF-β1)、炎性细胞因子和p53的mRNA表达水平,Western blotting检测p53蛋白的表达。结果显示,与对照组比较,酒精组小鼠肾质量、肾脏指数明显升高(P0.01);HE染色显示肾小球系膜细胞轻度增生,肾小管上皮细胞肿胀、间距增宽,间质内小血管增生、扩张、水肿,周围伴有炎性细胞浸润;PAS染色显示肾小球系膜细胞和系膜基质轻度增生;TGF-β1、炎性细胞因子(IL-1β、IL-6、TNF-α)及p53的mRNA表达增加(P0.05);p53蛋白表达下调(P0.01)。提示新型酒精性肾损伤小鼠模型构建成功,p53、TGF-β1及炎性细胞因子参与了新型酒精性肾损伤,这为今后的研究提供了可靠的模型和线索。     

IL-37及其与疾病相关性的研究进展

白细胞介素-37(IL-37)是一种新型炎性抑制因子,目前发现IL-37通过抑制促炎性细胞因子、趋化因子等的产生及对基因转录的调节,参与免疫和炎性反应,在多种炎性反应及免疫相关性疾病中起重要作用,为这些疾病提供新的治疗思路和新靶点。     

抗氧化剂对系膜细胞醛糖还原酶活性的影响

目的：观察抗氧化剂对高糖培养下肾小球系膜细胞(MC)内醛糖还原酶(AR)活性的影响，探讨抗氧化剂治疗糖尿病并发症的分子机制。方法:将牛肾小球系膜细胞培养于含高糖的培养基中3周，同时加入抗氧化剂过氧化物酶和DMSO，采用间接法观察抗氧化剂对细胞内醛糖还原酶活性的影响结果:高糖作用下系膜细胞内山梨醇含量增加，表明醛糖还原酶活性增高，而抗氧化剂能抑制醛糖还原酶活性的增高。结论:氧化应激和多元醇通路关系密切，抗氧化剂通过降低AR的活性和清除多元醇通路中产生的氧自由基而对糖尿病并发症有治疗作用，为临床运用尤其是早期运用抗氧化剂治疗糖尿病提供了理论依据。     

Sorbitol metabolism in inner medullary collecting duct cells of diabetic rats

Intracellular accumulation of sorbitol, generated fromd-glucose via the aldose reductase pathway, is thought to play an important role in diabetic complications such as lens cataracts and neuropathy. In order to elucidate the effect of diabetes on the renal inner medulla, another sorbitol-rich tissue, male Wistar rats were treated with a single dose of streptozotocin (60 mg/kg body weight, i.p.). Six wecks later total inner medullary tissue (IM) or isolated inner medullary collecting duct (IMCD) cells were prepared. In diabetic IM tissue, sorbitol content was 1.8-fold higher than in control IM tissue (134±17 vs. 74±22 mol/g tissue protein). Sorbitol production in both normal and diabetic IMCD cells was strongly dependent on extracellulard-glucose concentration. In normal cells, for example, sorbitol production was 90±9 mol sorbitol/g protein x h at 45 mMd-glucose compared to 13±1 mol/g protein x h at 5 mM. At identicald-glucose concentrations sorbitol synthesis in diabetic IMCD cells was, however, always significantly higher than in control cells (122% of control at 15 mM and 126% of control at 45 mM). In addition, aldose reductase activity in diabetic IM was found to be augmented. The maximal velocity was 4.2 times higher (97±22 U/g protein vs. 23±7 U/g protein) while theK _m of the enzyme remained unchanged. Membrane permeability for sorbitol or the response to changes in extracellular osmolarity was not significantly different in diabetic IMCD cells and normal cells with correspondingly high intracellular sorbitol concentrations. Similarly the kinetic parameters ofd-glucose uptake were not altered by streptozotocin treatment. These results suggest that increased medullary sorbitol content in diabetic rats is a result of increased sorbitol synthesis due to a higher extracellulard-glucose concentration and augmented aldose reductase activity in face of an unaltered sorbitol permeability of the plasma membrane.     

Regeneration and repair of myelinated fibers in sural-nerve biopsy specimens from patients with diabetic neuropathy treated with sorbinil

There is reason to believe that diabetic neuropathy may be related to the accumulation of sorbitol in nerve tissue through an aldose reductase pathway from glucose. Short-term treatment with aldose reductase inhibitors improves nerve conduction in subjects with diabetes, but the effects of long-term treatment on the neuropathologic changes of diabetic neuropathy are unknown. To determine whether more prolonged aldose reductase inhibition reverses the underlying lesions that accompany symptomatic diabetic peripheral polyneuropathy, we performed a randomized, placebo-controlled, double-blind trial of the investigational aldose reductase inhibitor sorbinil (250 mg per day). Sural-nerve biopsy specimens obtained at base line and after one year from 16 diabetic patients with neuropathy were analyzed morphometrically in detail and compared with selected electrophysiologic and clinical indexes. In contrast to patients who received placebo, the 10 sorbinil-treated patients had a decrease of 41.8 +/- 8.0 percent in nerve sorbitol content (P less than 0.01) and a 3.8-fold increase in the percentage of regenerating myelinated nerve fibers (P less than 0.001), reflected by a 33 percent increase in the number of myelinated fibers per unit of cross-sectional area of nerve (P = 0.04). They also had quantitative improvement in terms of the degree of paranodal demyelination, segmental demyelination, and myelin wrinkling. The increase in the number of fibers was accompanied by electrophysiologic and clinical evidence of improved nerve function. We conclude that sorbinil, as a metabolic intervention targeted against a specific biochemical consequence of hyperglycemia, can improve the neuropathologic lesions of diabetic neuropathy.     

The role of sorbitol pathway and treatment effect of aldose reductase inhibitor ONO2235 in the up-regulation of cardiac M2-muscarinic receptors in streptozotocin-induced diabetic rats

This study investigated the role of the sorbitol pathway on the genetic up-regulation of the cardiac M(2) muscarinic receptor (M(2)-mAChR) in streptozotocin (STZ)-induced diabetic rats. Three-month-old male Wistar rats were divided into five groups: (1) normal controls; (2) rats rendered diabetic by streptozotocin; (3) rats fed with glucose; (4) rats injected with sorbitol; and (5) diabetic rats treated with ONO-2235, an aldose reductase inhibitor. The M(2) muscarinic receptor (M(2)-mAChR) protein and mRNA densities of the heart tissue were measured by Western immunoblotting and Northern blotting, respectively. The densities of M(2)-mAChR protein and mRNA in the heart were significantly increased in diabetic rats, and rats given either glucose or sorbitol. When diabetic rats were treated with ONO-2235, the increases in heart M(2)-mAChR protein and mRNA were significantly reduced. The findings suggest that hyperglycemia and the sorbitol pathway are involved in the pathogenesis of diabetic heart disease in STZ-induced diabetic rats. Aldose reductase inhibitors may be useful in the treatment and prevention of cardiac complication in diabetes.     

Polyol Pathway Mediates Enhanced Degradation of Extracellular Matrix Via p38 MAPK Activation in Intervertebral Disc of Diabetic Rats

The aim of this study was to determine the significance of diabetes on degradation of intervertebral disc (IVD) extracellular matrix. Diabetic rats showed a significant increase in glucose and sorbitol contents in the IVD. The levels of aldose reductase, p38 and metalloproteinases, and degradation of metalloproteinase-derived aggrecan and type II collagen were increased, while tissue inhibitors of metalloproteinases levels were decreased in the IVD of diabetic rats. These changes were markedly affected by inhibition of aldose reductase or p38. Diabetes might contribute to enhanced matrix degradation in the IVD and the polyol pathway might mediate this process via p38 activation.     

DM2肾病患者血浆ET-1、血清TGF-β_1和尿Alb检测的临床意义

目的：探讨了2型糖尿病肾病（DN）患者血浆ET-1、血清TGF-β1和尿Alb水平的变化及临床意义。方法：应用放射免疫分析和酶联法对40例糖尿病无肾病和32例DN患者进行了血浆ET-1、血清TGF-β1和尿Alb进行了测定,并与35名正常健康人作比较。结果：DN组血浆ET-1、血清TGF-β1和尿Alb水平均显著地高于正常人组（P〈0.01）,NDN组与正常人组比较无显著性差异（P〉0.05）。结论：血浆ET-1、血清TGF-β1和尿Alb含量随DN的发生以及严重程度逐渐增高,可作为早期诊断DN发生以及严重程度的指标,对监测早期DN的发生和病情发展程度有重要的临床价值。     

2型糖尿病肾病患者血浆ET、血清TGF-β1和VEGF检测的临床意义

目的：探讨2型糖尿病肾病（DN）患者血浆ET、血清TGF-β1和VEGF水平的变化及临床意义。方法：应用放射免疫分析和酶联免疫吸附法对44例糖尿病无肾病患者和32例2型糖尿病肾病患者进行了血浆ET、血清TGF-β1和VEGF测定,并与35名正常健康人作比较。结果：DN组血浆ET、血清TGF-β1和VEGF水平显著高于正常人组（P〈0.01）。NDN组与正常人组比较无显著性差异（P〉0.05）。结论：血浆ET、血清TGF-β1和VEGF含量随DN的发生以及严重程度逐渐增高,可作为早期诊断DN发生以及严重程度的指标,对监测早期DN的发生和病情发展程度有重要的临床价值。     

Polymorphisms of sorbitol dehydrogenase (SDH) gene and susceptibility to diabetic retinopathy

The polyol pathway consists of two enzymes aldose reductase (AR) and sorbitol dehydrogenase (SDH); the former is the first enzyme in the polyol pathway, that catalyzes the reduction of glucose to sorbitol, the latter is the second one, that converts sorbitol to fructose using by NAD(+) as a cofactor. We along with others have recently found that SDH activity, the second step in the polyol pathway, might make a greater contribution to the etiology of diabetic retinopathy than does the first step involving AR. In this paper, we propose a novel hypothesis that polymorphisms of SDH gene may be correlated with SDH gene expression levels in diabetic retinas, thus being a valuable genetic marker for diabetic retinopathy.     

Nerve glucose, fructose, sorbitol, myo-inositol, and fiber degeneration and regeneration in diabetic neuropathy

We measured the alcohol sugars in sural nerves from 11 controls, 21 conventionally treated patients with diabetes and neuropathy, and 4 diabetics without neuropathy. The results were related to metabolic control and to clinical, neuropathological, and morphometric abnormalities in the nerves. The mean endoneurial glucose, fructose, and sorbitol values were higher in diabetic patients than in controls. Linear regression analysis revealed that nerve sorbitol content in the diabetics was inversely related to the number of myelinated fibers (P = 0.003). Mean nerve levels of myo-inositol were not decreased in the diabetic patients, with or without neuropathy, and were not associated with any of the neuropathological end points of diabetes. Our results indicate that myo-inositol deficiency is not part of the pathogenesis of human diabetic neuropathy, as had been hypothesized. Other accumulated alcohol sugars, however, were increased in diabetes and were associated with the severity of neuropathy. On repeat biopsy, six diabetics, treated for a year with the aldose reductase inhibitor sorbinil, had decreased endoneurial levels of sorbitol (P less than 0.01) and fructose (0.05 less than P less than 0.1), but unchanged levels of myo-inositol.