交感神经失能导致微血管形成障碍及功能受损引发糖尿病溃疡分子机制的研究

Previous studies have shown that the sympathetic nerve maintain skin homeostasis. when diabetic patients may also present sympathetic nerve failure, which reduces skin blood flow and impairs wound healing. However, diabetes mellitus induces poor wound healing of cutaneous lesions by a mechanism which is still unclear. Our previous studies showed that the sympathetic innervation of a variety of subunits within the skin, sympathetic domination in the wounds of diabetic mice full-thickness skin defects is significant differences with normal rats. We speculate that sympathetic dysfunctioPrevious studies have shown that the sympathetic nerve maintain skin homeostasis. when diabetic patients may also present sympathetic nerve failure, which reduces skin blood flow and impairs angioenesis and wound healing. However, Diabetes mellitus induces poor wound healing of cutaneous lesions by a mechanism which is still unclear. Our previous studies showed that microvascular formation is association with the process of wound healing of diabetic mice full-thickness skin defects; the sympathetic innervation of microvascular in the skin; The sympathetic nerve domination in the wounds of diabetic mice full-thickness skin defects is significant differences with normal rats. We speculate that sympathetic dysfunction may be an important a contributing factor on impairing wound healing of with diabetes mellitus, it impair angiogenesis and function of microvascular. In this study, we will firstly observe the molecular activity in the sympathetic on angioenesis via regulating vascular endothelial cells and pericytes of diabetic wound sites, establish a model of sympathetic nerve dysfunction of diabetic mice full-thickness skin defects healing. Thus access to the evidence that diabetes induced sympathetic damage, and then increase the microvascular damage lead to chronic ulcers. In future, for finding means which regulate sympathetic nerve function, improving the ability of healing in chronic wounds with diabetic ulcer. At the same time, It will help clarify the important role of sympathetic nerves in the skin wound healing, enrich tissue repair theory.n may be an important a contributing factor on impairing wound healing of with diabetes mellitus, it impair angiogenesis and dysfunction of microvascular. In this study, we firstly observe the molecular activity in the sympathetic regulation of diabetic skin. Thus access to evidence that diabetes induced sympathetic damage, and then increase the microvascular damage lead to chronic ulcers. In future, for finding means which regulate sympathetic nerve function, improving the ability of healing in chronic wounds with diabetic ulcer. At the same time, It will help clarify the important role of sympathetic nerves in the skin wound healing , enrich tissue repair theory.

既往研究表明，交感神经维持着皮肤的完整性，糖尿病患者出现交感神经损害会影响微小血管形成，减少皮肤血流造成愈合障碍。但是交感神经产生愈合困难的分子机制仍然不甚清楚。我们先前的研究显示，糖尿病鼠全层皮肤缺损的创面愈合过程中，微血管的形成受影响；皮肤中的微小血管受交感神经支配；而正常小鼠创面愈合过程中交感神经支配存在差异。因此，我们推测交感神经功能减退可能是影响糖尿病患者慢性溃疡难愈最重要的始动因素，它影响微血管的形成和功能是通过对血管内皮细胞与周细胞实现的。本课题首先观察交感神经递质对内皮细胞和周细胞生物学活动的影响作用；建立失交感神经的糖尿病鼠全层皮肤缺损的动物模型，观察愈合的情况。获取交感神经受损，进而引起糖尿病皮肤内微血管形成受损，功能发生障碍导致慢性溃疡的证据。为今后寻找控制交感神经功能，改善或促进糖尿病慢性创面愈合的手段。阐明交感神经在皮肤创面修复中的重要角色，丰富组织修复理论。

背景：糖尿病足普遍存在着上皮化缓慢、炎症延长、血管神经受损等问题，是糖尿病最严重的并发症之一。α-AR 阻断及α-AR、β-AR 同时阻断后内皮细胞变化尚不明确。普萘洛尔、酚妥拉明、6-OHDA、手术切除交感神经干作为多种的交感神经阻断或者切除的方式，在创面愈合中的作用和与血管形成的关系尚存在争议。.方法：将普萘洛尔、酚妥拉明分别或者同时处理人皮肤微血管内皮细胞，观察细胞增殖、毒性、划痕、成管。将普通大鼠切除腰交感神经干，观察对大鼠足部皮肤血管通透性和血管再生的变化，在大鼠骶尾部制作创面，观察腰交感神经干切除对大鼠骶尾部创面的影响。观察糖尿病小鼠和野生型小鼠皮肤结构差异和创面愈合差异。研究普萘洛尔乳膏外用、6-OHDA腹腔注射对糖尿病小鼠创面的影响。.结果：（1）普萘洛尔和酚妥拉明皆可单独或者协同抑制内皮细胞增殖、迁移、成管。（2）糖尿病小鼠和野生型正常小鼠皮肤组织结构和交感功能有较大差异。（3）正常小鼠创面普萘洛尔口服组愈合速度快于对照组。（4）糖尿病小鼠创面普萘洛尔外用组的愈合速度快于对照组。普萘洛尔组IL-1β、ang2 表达低于对照组。（5）在交感神经切除后骶尾部和后足的皮肤可出现去交感，但一段时间恢复。（6）腰交感神经切除术加速骶尾区创面收缩。（7）在腰交感神经干切除后大鼠后足底皮肤伊文思蓝的浓度显著下降，腺苷A2A受体表达显著降低。（8）在糖尿病小鼠6-OHDA处理组创面收缩快于对照组，但表皮愈合慢于对照组。NE、EGF、IL-1β、NG2、desmin和Ang-2在实验组的表达均低于在对照组。Ang-1实验组较对照组表达量高。.结论：普萘洛尔和酚妥拉明抑制内皮细胞增殖、迁移、成管。糖尿病小鼠皮肤与正常皮肤不同，在全身化学去交感后出现了创面收缩加快，上皮化延迟，炎症反应和血管再生受抑制。外用普萘洛尔可能成为治疗糖尿病溃疡的一种新的手段。手术切除腰交感神经干后可造成局部的失交感，并选择性抑制血管再生和通透性。