负压创面治疗技术对大鼠创面血管生成的影响及分子机制的研究

发布时间：2018-04-05 16:38

本文选题：负压创面治疗　切入点：糖尿病　出处：《第四军医大学》2013年硕士论文

【摘要】：研究背景及目的随着社会发展及人口老龄化，日常生活中急慢性创面的发生率也明显增加。车祸或自然灾害等急性创伤因素往往导致皮肤软组织缺损、肌腱或骨质外露。这类创面的修复在临床往往比较复杂，是临床医师面临的挑战之一。临床常规的治疗方法是经创面准备使创面形成健康的肉芽组织后行手术植皮或皮瓣移植来修复创面。湿敷换药是最常用的创面准备方法，通过创面湿敷换药使创面形成新鲜的肉芽组织，提高植皮及皮瓣移植手术的成功率。但常规湿敷换药要达到满意的创基准备要求需要耗费较长的时间并且往往因为频繁换到给患者带来不可避免的痛苦。对于一些伴有肌腱暴露的创面，简单的湿敷换药往往不能达到良好的创基准备要求。负压创面治疗技术（Negative Pressure Wound Therapy）又称真空辅助闭合疗法（VAC，Vacuum assisted closure），是近年来兴起的一种促进创面愈合的前沿技术并被广泛应用于临床各种创面的治疗，尤其是一些复杂难愈性创面的治疗。负压创面治疗技术的概念是利用医用多聚泡沫敷料及半透生物贴膜封闭创面后，利用引流管将创面敷料与负压装置连接达到治疗创面的目的。既往研究表明负压可以通过增加创面血流、促进创面肉芽组织生长，减少细菌定植和繁殖及保持伤口环境湿润。虽然NPWT已被广泛应用，但其作用的详细分子机制目前仍不明确，尤其是关于其对创面血管新生过程的影响及机制的研究比较罕见。因此本课题拟以正常大鼠及糖尿病大鼠后背全层皮肤缺损创面为模型从功能上及组织病理结构上验证NPWT对创面血管生成过程的影响，并探讨部分分子机制。方法 1.雄性SD大鼠40只，随机分为治疗组和对照组。大鼠后背正中偏上1cm处创建2cm×2cm大小全层皮肤缺损创面。对照组创面用医用泡沫敷料包扎予以常规换药，治疗组创面用医用泡沫敷料覆盖后，每天予以4小时120mmHg的持续负压治疗，连续给予负压治疗7天。分别在术后1、3、7天检测创面血液灌注情况，利用HE及免疫组化染色观察创面血管生成状况，PCR技术检测创面组织血管形成调节因子的表达情况。 2.雄性SD大鼠60只，，随机分成正常大鼠组和糖尿病大鼠组。糖尿病组大鼠经腹腔单次注射链脲佐菌素诱导糖尿病模型，注射后48小时检测大鼠尾静脉血糖浓度，血糖浓度＞16.7mmol/L视为糖尿病模型建立。两周后在两组大鼠后背正中创建全层皮肤缺损模型，创面予以常规换药。观察两组大鼠生活习性及大体表观的变化。分别在术后1、3、7、14天计算比较两组大鼠创面愈合率的差异。利用激光多普勒技术分别在术后1、3、7天检测创面血流灌注状况。 3.60只链脲佐菌素诱导的糖尿病大鼠，随机分成治疗组及对照组，在后背创建全层皮肤缺损创面。对照组创面予以常规换药，对照组创面用医用泡沫敷料覆盖后，每天予以120mmHg持续负压治疗4小时，治疗维持7天。分别在术后1、3、7天观察创面血液灌注情况，创面组织做病理切片观察创面血管形成状况，利用PCR技术检测创面血管形成调节因子的表达情况。结果 1.与对照组相比，NPWT明显增加了创面血流灌注量（P＜0.05）。创面血管密度在术后第3天明显高于对照组（P＜0.05），术后第7天，NPWT治疗组创面新生血管排列规律，管腔宽畅，而对照组排列紊乱，管腔迂曲。治疗组创面血管生成的正性调节因子VEGF及Ang1在术后第3天明显高于对照组（P＜0.05）。 2.与正常大鼠比较，糖尿病大鼠多饮、多尿及多食症状明显，皮毛发黄，体重增加缓慢。创面愈合率在术后第3、7、14天明显低于正常大鼠（P＜0.05）。创面血液灌注量术后第1、3、7天明显低于对照组（P＜0.05）。 3.治疗组创面血流灌注量在各时间点均高于对照组。术后第3天，治疗组创面血管密度明显高于对照组（P＜0.05），第七天创面血管密度在治疗组及对照组间无明显差别，但组织病理显示治疗组创面新生血管排列规律，管腔宽大通畅；术后第7天及14天创面血管形成负性调节因子Ang2在治疗组明显低于对照组（P＜0.05）。结论 1. NPWT可以增加正常大鼠创面血流灌注，有益于创面血管生成过程。NPWT促进创面血管生成的机制可能与上调促血管生成因子VEGF及Ang1的表达有关。 2.糖尿病大鼠创面血供不良，创面血管生成功能障碍可能是其创面难愈的原因之一。 3. NPWT可以改善糖尿病创面血液灌注，增加创面早期新生血管密度，有利于新生血管的成熟。对血管生成素及其受体通路的影响也是其可能的机制之一。
[Abstract]:Background and purpose of research
With the social development and population aging, the incidence of acute and chronic wounds in daily life also increased significantly. The accident or natural disasters and other factors often lead to acute traumatic skin and soft tissue defect, tendon or bone exposure. This type of wound repair in clinic is often more complex, is one of the challenges facing clinicians. Conventional treatment the method is to make the clinical wound wound to form a healthy granulation tissue were surgical skin grafting or flap to repair the wound. The wound wet dressing is the most commonly used preparation methods, formation of fresh granulation tissue through the wound wet dressing the wound, skin flap transplantation and improve the success rate of the operation. But the conventional wet dressing to achieve with the initial preparation requires longer time and often because of frequent change to bring patients the inevitable pain. For some with tendon exposed wound The surface, simple wet dressing often cannot achieve good initial preparation requirements.
Negative pressure wound therapy (Negative Pressure Wound Therapy) is also called vacuum assisted closure therapy (VAC Vacuum, assisted closure), is on the rise in recent years a cutting-edge technology to promote wound healing and is widely used in all kinds of clinical wound, especially in the treatment of some complicated wounds. The concept of negative pressure wound therapy is the use of medical poly foam dressing and semi permeable biological film wound closure, the drainage tube will wound dressings and negative pressure device is connected to the wound treatment. Previous study showed that negative pressure can increase blood flow through the wound, promote the growth of granulation tissue, reduce the bacterial colonization and reproduction and keep the wound moist environment. Although NPWT has been with wide application, but its molecular mechanism is still not clear, especially about its effects on wound angiogenesis and mechanism research Relatively rare. Therefore, we aim to verify the effects of NPWT on wound angiogenesis in normal rats and diabetic rats, and to explore some molecular mechanisms.
Method
1. of 40 male SD rats were randomly divided into treatment group and control group. The rats back at 1cm 2cm to create a partial median size of 2cm * full-thickness skin wounds. The wounds of control group with medical foam dressing treatment group received conventional dressing, wound covered with medical foam dressing, every day to 4 continuous negative pressure 120mmHg hours of treatment, continuous treatment for 7 days respectively. Given the negative pressure in 1,3,7 days after detection of wound blood perfusion, using HE and immunohistochemical staining to observe the wound angiogenesis, vascular wound tissue detection PCR technique to form expression regulator.
2. of 60 male SD rats were randomly divided into normal group and diabetic rats group. Diabetic rats were induced by a single intraperitoneal injection of streptozotocin induced diabetic model of rats was detected 48 hours after injection of tail vein blood glucose concentration, glucose concentration, 16.7mmol/L as the diabetic model established after two weeks. In the back of the two groups of rats was created full-thickness skin defect model, wound dressing change. To observe two groups of rats living habits and generally apparent changes in 1,3,7,14 days after the operation respectively. Comparing the two groups of rats wound healing rate difference. Using laser Doppler technique in 1,3,7 days after detection of wound blood perfusion status.
Only 3.60 of diabetic rats induced by streptozotocin, were randomly divided into treatment group and control group, create full-thickness skin wounds in the back. The control group was given routine treatment, the control group was covered with medical foam dressing, every day to 120mmHg 4 hours of continuous negative pressure therapy, treatment lasted for 7 days. In operation after 1,3,7 days to observe the wound blood perfusion, wound tissue pathological section observation of wound angiogenesis, vascular wound detection using PCR technique to form the expression of regulatory factors.
Result
1. compared with the control group, NPWT significantly increased the wound blood perfusion (P < 0.05). Vascular density in third days after operation was significantly higher than the control group (P < 0.05), seventh days after operation, NPWT group wound angiogenesis arrangement rule of lumen spacious, while the control group arranged disorderly, lumen tortuous. The treatment group was wound angiogenesis regulatory factor VEGF and Ang1 in third days after operation was significantly higher than the control group (P < 0.05).
2., compared with the normal rats, diabetic rats had more symptoms of polyuria, polyuria and polyarthritis. Their hair was yellow and their weight gain was slow. The wound healing rate was significantly lower than that of the normal rats at 3,7,14 days after operation (P < 0.05). The blood perfusion volume of the wound was significantly lower than that of the control group at 1,3,7 days after operation (P < 0.05).
3. wound treatment group blood perfusion were higher than the control group at each time point. Third days after operation, wound treatment group vascular density was significantly higher than the control group (P < 0.05), seventh days of wound vascular density had no obvious difference between the treatment group and the control group, but the pathological change showed that treatment group wound neovascularization arrangement rule after seventh large lumen patency; and 14 days wound angiogenesis negative regulatory factor Ang2 in treatment group was significantly lower than the control group (P < 0.05).
conclusion
1. NPWT can increase the blood perfusion of normal rats, which is beneficial to the angiogenesis of wounds. The mechanism of.NPWT promoting angiogenesis is probably related to the up regulation of angiogenesis factors VEGF and Ang1 expression.
2. wound blood of diabetic rats for poor wound angiogenesis dysfunction may be one of the reasons for the wound healing.
3. NPWT can improve the blood perfusion of diabetic wound and increase the early neovascularization of the wound, which is beneficial to the maturation of new blood vessels. The effect of angiopoietin and its receptor pathway is also one of its possible mechanisms.

【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R641

【参考文献】