MMP-9/TIMP-1表达失衡与糖尿病大鼠皮肤"隐性损害"关系的初步探讨

目的 观察基质金属蛋白酶9(MMP-9)、组织金属蛋白酶抑制物1(TIMP-1)的表达和MMP-9/TIMP-1比值在糖尿病组和正常组大鼠皮肤表达的变化,并探讨其可能的作用.方法 采用链脲佐菌素制备糖尿病大鼠模型,6周后采用HE染色、免疫组织化学方法评估皮肤组织的组织学、细胞生物学行为的情况;通过逆转录-聚合酶链反应(RT-PCR)和Western印迹检测两组大鼠皮肤MMP-9、T1MP-1的表达情况.结果 与对照组比较,糖尿病组皮肤表皮厚度变薄,皮肤组织层次欠清晰、缺乏复层排列、明显萎缩,真皮纤维束排列紊乱、纤维间距增加、炎性细胞浸润.糖尿病组皮肤MMP-9的表达高于正常组,而TIMP-1的表达低于正常组,MMP-9/TIMP-1蛋白水平比值高于正常组.结论 糖尿病鼠皮肤在组织结构完整性未遭到破坏的情况下已经存在组织学、细胞生物学行为的改变,这种"隐性损害"可能与MMP-9/TIMP-1的平衡改变有关.     

线粒体功能障碍对糖尿病心肌病发病进展的影响

近年来糖尿病的发病率呈上升的趋势,心血管疾病是糖尿病患者常见的一个并发症,它也是导致糖尿病患者高死亡率的一个重要因素.糖尿病心肌病是指患者在排除了高血压、冠心病等疾病后由单纯高血糖所致的心脏功能及结构改变.糖尿病患者的心肌常存在着代谢功能的异常,线粒体是细胞能量代谢的中心,最近有研究表明,线粒体功能异常在糖尿病心肌病的进展过程中可能起到了至关重要的作用.本文将对近年来关于线粒体功能异常对糖尿病心肌病的影响及其可能的机制进行综述.     

糖基化基质对人皮肤成纤维细胞增殖凋亡平衡的研究

目的探讨糖尿病皮肤细胞外基质（ECM）糖基化改变与细胞增殖凋亡平衡的相关性。方法DM或非DM患者的皮肤组织和溃疡创面标本组织学观察,同时免疫组化法检测皮肤组织增殖细胞核抗原（PCNA）、凋亡细胞、糖基化终末产物（AGE）和糖基化终末产物受体（RAGE）表达;体外建立糖基化基质,RAGE抗体干预后,检测细胞存活率、细胞周期和凋亡。结果与对照组相比,DM皮肤胶原萎缩,凋亡细胞比例增高;创面肉芽形成不良,细胞增殖能力减弱,凋亡细胞增加,AGE和RAGE表达增多;体外糖基化基质上接种成纤维细胞存活率明显减少,细胞周期运行障碍,凋亡增加,阻断AGE-RAGE间相互作用可基本缓解糖基化基质对细胞增殖凋亡过程的影响。结论DM皮肤组织中糖基化ECM蓄积是细胞功能改变的重要环境介质,经RAGE介导影响细胞增殖凋亡平衡,致糖尿病皮肤创伤起点异常,创面难愈。     

外阴皮肤黏膜特征性改变预诊糖尿病40例临床观察体会

目的总结外阴皮肤黏膜特征性改变对于糖尿病发生的预示价值,以在了解患者特征的基础更好的实现糖尿病预防以及早期治疗,最大程度上改善患者的生活质量。方法该次实验研究的患者均为该院在2013年1月—2014年5月期间收治治疗的外阴痒痛异常、感觉异常且反复发生外阴阴道白色念珠菌感染的患者,总计40例,对这些患者的临床治疗进行回顾性分析。对40例患者进行详细的临床检查,最终发现患者存在明显性的外阴皮肤黏膜特征性改变,这些患者经过至少两种方法排除了糖尿病问题。结果该次实验研究的40例患者,在1~2年后最终均被确诊为糖尿病。结论外阴皮肤黏膜特征性改变也可以视为糖尿病性改变,有明显区别于外阴疾病的皮肤黏膜改变,属于典型的外阴皮肤黏膜改变。在对患者进行妇科检查时,要求医师必须注重观察患者的外阴皮肤黏膜情况,了解患者是否已经提前发生了糖尿病性改变问题,在明确是糖尿病发病前的预示标准后,对患者进行提前的预防治疗,控制糖尿病的发生几率,或者最大程度上缓解糖尿病的发生,保证患者生活质量。     

糖尿病肺损伤的研究进展

研究表明,肺是糖尿病攻击的靶器官之一,其损害主要涉及肺功能及病理学改变,糖尿病患者肺功能损害主要表现为通气功能障碍、小气道功能减退和弥散功能异常,而糖尿病肺组织病理学改变主要表现为基底膜增厚和细胞外基质增生以及超微结构的改变.积极控制血糖水平及胰岛素等药物治疗可能改善糖尿病患者的肺功能.深入研究糖尿病肺部病变,有可能在防治糖尿病血管并发症方面提出新的思路.     

糖尿病大鼠磷酸化表皮生长因子受体表达的改变及其与隐形皮肤损害的关系

目的探讨糖尿病大鼠皮肤的隐形损害现象,以及磷酸化的表皮生长因子受体（P-EGFR）在糖尿病皮肤中的表达与这种隐形损害的关系。方法对比观察不同病期的糖尿病大鼠模型背部和尾巴皮肤组织学改变以及超微结构变化,用免疫组化及蛋白印迹的方法检测P-EGFR在糖尿病大鼠皮肤中的表达。结果糖尿病大鼠皮肤存在组织学和超微结构的改变,p-EGFR在皮肤组织中的表达下降。结论P-EGFR在糖尿病大鼠皮肤组织中表达下调可能与糖尿病皮肤损害的发生有关。     

糖尿病患者80例心电图分析

目的 探讨糖尿病患者的临床心电图表现特征.方法 对80例糖尿病患者的心电图资料进行分析,80例性别、年龄匹配的健康体检者的心电图资料作为对照.结果 糖尿病患者心电图异常改变33例(41％),正常对照组心电图异常改变8例(10％),二者差异有统计学意义(P＜0.005).糖尿病患者心电图改变以ST-T改变、无痛性心肌梗死、心律失常为主.结论 糖尿病患者心电图异常发生率较高,女性高于男性,多表现为ST-T改变、无痛性心肌梗死、心律失常.     

调节性B细胞与1型糖尿病

1型糖尿病主要是由T细胞介导的针对胰岛β细胞的自身免疫性疾病,然而B细胞在1型糖尿病发生、发展中也发挥重要作用,B细胞去除可用于治疗1型糖尿病.调节性B细胞是近来研究发现的一群具有免疫负向调控作用的B细胞亚群.在1型糖尿病中存在调节性B细胞数量和功能的异常.探索调节性B细胞在1型糖尿病发病机制中所起的作用将提供更多的理论依据,为1型糖尿病免疫治疗提供新靶点和新思路.     

糖尿病对大鼠皮肤角质形成细胞功能损害的影响

目的 探讨糖尿病对大鼠皮肤角质形成细胞生物学行为的影响.方法 将SD大鼠随机分为糖尿病组和对照组,诱导糖尿病大鼠模型,并制作深Ⅱ度烫伤大鼠模型,测定伤后3、7、14 d和21 d的创面愈合率;观察两组表皮组织的组织学特征及厚度,测定两组角质形成细胞贴壁率、细胞周期、早期凋亡率、迁移能力,观察晚期糖基化终末产物(AGEs)蓄积情况.结果 糖尿病组与对照组比较,在伤后7、14 d和21 d创面愈合面积百分比显著减少(P<0.05).糖尿病组皮肤表皮细胞层次欠清晰,部分表皮细胞缺乏复层排列,细胞数量明显减少;糖尿病组角质形成细胞12、24 h贴壁率显著下降(P<0.05),进入G2/M期的细胞显著减少(P<0.05),早期凋亡细胞的比例显著增高(P<0.05),细胞迁移能力明显低于对照组.糖尿病大鼠表皮层中有大量的AGES蓄积.结论 糖尿病大鼠创面愈合过程中存在再上皮化受阻,这一现象与表皮角质形成细胞的生物学功能受抑有关.表皮层大量AGEs蓄积可能与糖尿病环境下角质形成细胞牛物学功能受抑有关.     

糖尿病视网膜病变细胞外间质和粘附分子的研究进展

细胞外间质 (ECM)、粘附分子 (AMs)及生长因子 (GFs)与糖尿病微血管病变有密切的关系。研究不同阶段糖尿病视网膜病变在基膜、细胞表面等部位ECM、AMs的异常改变有助于了解其发病机制。ECM、AMs成分的改变和交联异常与非酶糖基化导致的糖基化终末产物有关 ,而许多GFs能诱导和促进此过程的发生和发展。血液ECM成分和AMs的含量测定对于糖尿病微血管病变的早期诊断、病情监测、疗效观察和预后判断都不失为良好的指标。     

Leprdb diabetic mouse bone marrow cells inhibit skin wound vascularization but promote wound healing

Bone marrow stem cells participate in tissue repair processes and may have roles in skin wound repair. Diabetes is characterized by delayed and poor wound healing, and type 1 diabetes seems to lead to stem cell dysfunction. Hence, stem cell dysfunction could contribute to poor healing, and stem cell-based therapies may be efficacious in diabetic wounds. We investigated the potential of exogenous stem cells to promote skin healing and possible effects of type 2 diabetes on stem cell function. Mouse bone marrow cells from nondiabetic and diabetic mice were enriched for putative stem cells and injected under skin wounds of nondiabetic or type 2 diabetic Leprdb mice. Using histology and morphometry, vascularization and healing in treated and untreated mice were analyzed. We anticipated a correlation between improved wound healing and vascularization, because therapies that increase tissue vascularization tend to enhance wound healing. Our data indicate that exogenous nondiabetic bone marrow-derived cells increase vascularization and improve wound healing in Leprdb mice but have little effect on nondiabetic controls. In contrast, Leprdb-derived marrow cells inhibit vascularization but promote wound healing in Leprdb mice. Thus, adult stem cell function may be impaired by type 2 diabetes; the ability to promote vascularization and wound healing are distinct functions of bone marrow cells; and neovascularization and wound healing may not be tightly coupled. Additionally, we observed little incorporation of injected cells into wound structures, suggesting that improved healing is mediated through mechanisms other than direct differentiation and incorporation of the cells.     

Dicer expression is impaired in diabetic cutaneous wound healing

Diabetes, as a fast growing non-communicable disease, is one of the major health problems of the twenty-first century. Several complications such as cardiovascular disease and renal failure are accompanying diabetes. The chronic cutaneous wound is another diabetes complication, results from the reduced body healing potential. At genome level, diabetic wound environment displays disorganized gene functions emphasizing the critical role of gene regulatory networks in the control of chronic wound repair. MicroRNAs, major regulators of gene activity, have been shown to be impaired in several pathological conditions such as chronic wounds. A reason behind that can be sought in the impaired activity of enzymes involved in the development and production of miRNAs. In current study, streptozocin-induced diabetic rats and non-diabetic controls were used to study the effect of diabetes on Dicer presence in wound environment. Unwounded skin of diabetic animals showed significantly lower level of Dicer expression compared with non-diabetic animal-derived skin. However, at day 7 post-wounding, diabetic animal-derived wounds contained a higher level of Dicer expression compared with non-diabetic ones. Parallel to these findings, the granulation tissue formation and wound closure are impaired in diabetic wounds. This study highlights the dysregulated presence of Dicer at different stages of the diabetic cutaneous wound.     

Circ-Klhl8 overexpression increased the therapeutic effect of EPCs in diabetic wound healing via the miR-212-3p/SIRT5 axis

Previous studies found that hypoxic pretreatment of endothelial progenitor cells (EPCs) prior to transplantation had a greater therapeutic effect than untreated EPCs in promoting diabetic wound healing. However, the exact mechanism is uncertain. Here, circRNA expression in EPCs after hypoxic treatment was investigated. High-throughput sequencing was used to assess abnormal expression by EPCs of circular RNAs (circRNAs) following hypoxic pretreatment. Additionally, an in vivo full-thickness skin defect mouse model was used to assess the effects of transplanted EPCs on diabetic wound closure. Subsequently, the regulatory mechanism and targets were studied. The results showed that circ-Klhl8 overexpression suppressed hyper glucose-induced endothelial cell damage by activating autophagy. MiR-212-3p and SIRT5 were identified as the downstream targets of circ-Klhl8. Circ-Klhl8 overexpression promoted skin wound healing by regulating SIRT5-mediated autophagy. In conclusion, the study found that circ-Klhl8 overexpression increased the EPC therapeutic effect in promoting diabetic wound healing by targeting the miR-212-3p/SIRT5 axis.     

CD34+ blood cells accelerate vascularization and healing of diabetic mouse skin wounds

Diabetes is characterized by poor circulation and impaired angiogenesis, which appear to contribute to the frequent skin lesions and poor wound healing common in diabetic patients. Therapies to improve circulation commonly improve wound healing in diabetic patients. Administration of circulating CD34+ cells, cells that can function as endothelial cell progenitors, accelerates blood flow restoration to ischemic limbs of diabetic mice. We have investigated the potential of these cells to accelerate revascularization and healing in full-thickness skin wounds of hypoinsulinemic (streptozotocin-treated) diabetic mice. Wounds were injected with human CD34+ or CD34- peripheral blood mononuclear cells or no cells, and analyzed for vascularity and healing at various times thereafter. Treatment with CD34+ enriched cells decreased wound size by 4 days after treatment, accelerated epidermal healing, and rapidly and dramatically accelerated revascularization of the wounds compared to controls. Initially increased vascularization was mediated principally by an increase in vessel diameter, but later, both an increase in vascular size and number were observed. These findings indicate that blood-derived progenitors may have therapeutic potential in the treatment of skin lesions in the setting of diabetes, and give insights into how bone marrow cells exert their effects on neovascularization.     

Wnt/β-catenin信号通路在糖尿病创面难愈中的作用

目的 探讨Wnt/β-catenin信号途径在糖尿病创面难愈中的作用.方法 在1型糖尿病大鼠制作背部皮肤缺损,分为对照组、糖尿病组、氯化锂组和表皮生长因子(EGF)组,观察背部创面愈合情况及β-catenin表达变化.结果 伤后各组大鼠创面未见感染征象.对照组、氯化锂组及EGF组大鼠与糖尿病组大鼠相比,创面愈合时间短,创面愈合率高,伤腔容积缩小显著,肉芽组织成熟,且β-catenin阳性细胞率高,差异有统计学意义(P＜0.05或P＜0.01).结论 Wnt/β-catenin信号通路参与了糖尿病大鼠创面愈合过程.

Abstract：

Objective To investigate the role of Wnt/β-catenin signaling pathway in impaired wound healing of diabetes mellitus.Methods The back skin defect was produced in rats with type1diabetes.All of these rats were divided into normal group, diabetes group, lithium chloride group, and epidermal growth factor (EGF) group.The back wound healing and β-catenin expression were observed.Results There were no signs of infection in the wound of rats after injury.Compared with diabetic group, the wound healing time was shorter,wound healing rate was higher, wound cavity volume was smaller, granulation tissue was more mature, and β-catenin positive cell rate was higher in normal group, lithium chloride group, and EGF group(P＜0.05 or P＜0.01).Conclusions Wnt/beta-catenin signaling pathway is involved in the process of wound healing in diabetic rats.     

神经肽P物质在糖尿病足溃疡皮肤中的表达变化

目的 探讨神经肽P物质（SP）在糖尿病足溃疡（DFU）皮肤中的表达变化. 方法 免疫组织化学法观察对照（Con）、非糖尿病DFU（NDFU）组和DFU组皮肤组织中SP及增殖细胞核抗原（PC-NA）的表达特点. 结果 SP表达量在Con、NDFU和DFU组分别为79.64、65.12和45.30 （P＜0.01）.PCNA阳性细胞率在Con、NDFU和DFU组分别为（35.120±7.314）％,（33.880±4.055）％和（28.320±9.332）％,NDFU组与Con、DFU组比较差异有统计学意义（P＜0.01）,但NDFU组和Con组比较差异无统计学意义（P＞0.05）. 结论 DFU患者边缘皮肤中SP表达量减少,可能是导致糖尿病患者创面炎症反应异常及创面修复细胞增殖功能受损,从而引起创面修复受损的原因之一.     

L-Arginine supplementation enhances diabetic wound healing: involvement of the nitric oxide synthase and arginase pathways

Diabetes impairs wound healing and there are few therapeutic options to reverse it. Previous work has demonstrated the importance of nitric oxide for successful wound healing. In diabetes, NO synthesis is reduced in the wound milieu. The amino acid L-arginine is the only substrate for NO synthesis. We hypothesized that L-arginine supplementation would enhance wound healing by restoring NO synthesis. Thirty-six male Sprague-Dawley rats (body weight, 225 to 250 g) were separated in 4 groups: 20 rats were rendered diabetic 7 days prior to wounding by intraperitoneal streptozotocin (STZ) injection (70 mg/kg). Sixteen rats served as controls. Half of the animals of each group received 1 g/kg supplemental L-arginine administered by gavage twice daily. Control rats were gavaged with water. Treatment was started 3 days before wounding. All rats underwent a dorsal skin incision and subcutaneous implantation of polyvinyl alcohol (PVA) sponges. The rats were killed 10 days post wounding and wound breaking strength, hydroxyproline content of the sponges, nitrite/nitrate (NO(x)) concentration, arginase activity, and amino acid composition of the wound fluid and plasma were analyzed. Wound fluid NO(x) concentrations and wound breaking strength were significantly reduced in the diabetic group compared to the controls. L-Arginine treatment restored diabetic NO(x) levels toward normal values and significantly enhanced wound breaking strength. Wound fluid arginase activity and ornithine concentrations were significantly lower in the diabetic animals but unaffected by treatment. The data demonstrate that the impaired NO synthesis in the diabetic wound milieu can at least partially be reversed by arginine supplementation. In view of previous results on the importance of NO for wound healing, the data suggest that arginine supplementation restores impaired healing in this acute wound model by normalizing the NO pathway but without affecting arginase activity.     

糖尿病周围神经病700例临床与神经电生理分析

目的探讨糖尿病周围神经病的临床和电生理特点,明确电生理检查的诊断价值。方法对700患者进行感觉和运动神经传导测定,240例患者进行针极肌电图测定。结果507例(724%)患者电生理检查异常,其中307例(606%)为多发性周围神经病,74例(146%)为腕管综合征;感觉神经传导异常程度重于运动神经,波幅的下降程度较传导速度减慢明显,下肢重于上肢(P<005)。仅有46%的患者针极肌电图异常而神经传导正常。结论糖尿病周围神经病的临床和电生理表现均以感觉神经受损为主;电生理检查有助于发现临床病变,但并非所有患者均能发现电生理异常;建议不将针极肌电图进行糖尿病周围神经病的筛查作为常规使用。     

Improved refractory wound healing with administration of acidic fibroblast growth factor in diabetic rats

The aim of the present study is to investigate the effect and mechanism of acidic fibroblast growth factor (aFGF) on treating refractory wound of diabetic rats. SD rats were randomly divided into control group, diabetes group, and aFGF group. Ulcer skin tissues of three groups of rats were respectively collected on days 7 and 14 after establishment of ulcer model for biochemical test, pathological section and immunohistochemistry to comprehensively evaluate the treatment effect of aFGF on diabetic ulcer. The results showed that aFGF could significantly increase capillaries and fibroblast amounts of ulcer tissues, enhance the expression of TGF-β and PCNA proliferation proteins, and thus improved diabetic ulcer tissues. The preliminary mechanism that aFGF helps to promote healing of diabetic ulcer is possibly associated with that aFGF stimulated ulcer skins to secrete TGF-β and PCNA proteins and promoted proliferation of capillaries and fibroblasts.