荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

目的 观察荷负电气溶胶治疗对大鼠烫伤创面愈合过程中白细胞介素(IL)-8、IL-10表达的影响,探讨荷负电气溶胶治疗促进创面愈合的作用机制.方法 制作SD大鼠深Ⅱ度烫伤模型,采用分组对照方法,将40只鼠随机分为治疗组(n1=20)和对照组(n2=20).治疗组应用荷负电气溶胶治疗,每次1.5 h,每天2次,直至创面愈合;对照组不作荷负电气溶胶治疗.伤后第1～11天分别取创面标本制作切片,采用免疫组织化学和图像分析方法,检测创面愈合过程中IL-8和IL-10表达水平.结果 创面平均愈合时间治疗组为(7.00±1.15)d,对照组为(9.00±1.34)d,治疗组创面愈合时间明显提前(P＜0.01).免疫组织化学显示,两组IL-8均在伤后第1天开始表达.主要位于多核粒细胞和单核细胞;第3天表达明显增多达高峰,并见大量成纤维细胞表达,治疗组的峰值明显低于对照组,差异有统计学意义(P＜0.01),第5～11天表达水平迅速下降.两组IL-10伤后第1天在淋巴细胞和单核细胞均有表达;第3天开始有角质细胞表达并达高峰,第5～11天表达水平缓慢下降,但治疗组要明显高于对照组,第3～11天差异有统计学意义(P＜0.01).结论 荷负电气溶胶治疗能有效抑制创面IL-8的表达及促进IL-10的表达,缩短炎症进程,从而加速创面愈合.

Abstract：

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

解毒烧伤膏对深Ⅱ度烫伤创面愈合中白细胞介素8、10表达的影响

解毒烧伤膏能不同程度地促进浅Ⅱ、深Ⅱ度烧伤和残余创面的愈合，无明显不良反应。为进一步探讨其在促进创面愈合过程中的分子机制，笔进行了下述实验。     

荷负电气溶胶治疗Ⅱ度烧伤创面的临床效果及病理学观察

目的观察荷负电气溶胶(下称气溶胶)治疗Ⅱ度烧伤创面的效果。方法选择单纯浅Ⅱ、深Ⅱ度烧伤患者,随机分为:(1)气溶胶组:浅Ⅱ度180例、深Ⅱ度100例,伤后6h~2d开始用气溶胶治疗创面,l~2次/d,1.5h/次。(2)对照组:浅Ⅱ、深Ⅱ度患者各30例,常规治疗。(3)自身对照组:浅Ⅱ、深Ⅱ度患者各10例,同上用气溶胶治疗,但同一患者部分创面覆盖无菌金属片屏蔽气溶胶(屏蔽组),部分创面不屏蔽(非屏蔽组)。观察气溶胶治疗过程中患者创面的大体变化,治疗前后进行创面细菌培养,并监测其肝、肾功能及血生化指标有无改变。记录各组患者创面愈合时间。另制作深Ⅱ度烫伤大鼠模型,同前分为气溶胶组和对照组并治疗。取两组大鼠治疗前及治疗后1、2、3周的创面组织标本,作病理学观察。结果气溶胶治疗后患者创面渗出少,治疗前后均无细菌生长。总体来讲,气溶胶治疗前后患者肝、肾功能及血生化指标无明显改变。气溶胶组患者浅Ⅱ度创面伤后(6.3±1.6)d愈合,深Ⅱ度创面(15.1±3.1)d愈合,明显短于对照组相同深度创面[(11.3±1.4)、(21.2±1.4)d,P<0.01]。自身对照组中,相同烧伤深度的非屏蔽组与屏蔽组比较,创面愈合时间也明显缩短(P<0.01)。病理学检查显示,气溶胶组大鼠治疗后第3周皮肤结构已基本恢复正常,而对照组此时恢复较差。结论气溶胶能有效促进Ⅱ度烧伤创面的愈合且使用安全。     

荷负电气溶胶促进烧伤患者头部取皮区创面愈合效果观察

目的探讨荷负电气溶胶促进烧伤患者头部取皮区创面愈合的效果。方法将120例患者随机分为气溶胶组（61例）和对照组（59例），两组均采用电动取皮机取头皮，厚度0．25mm。取皮后对照组采用常规处理；气溶胶组采用气溶胶对头部取皮区创面进行治疗护理，每次2h，每天2次，直至创面愈合。结果两组创面疼痛消失、创面渗出物消失情况及愈合时间比较，差异有显著性意义（P〈0.05、P〈0.01）。结论荷负电气溶胶能有效促进烧伤患者头部取皮区创面愈合。     

海水浸泡对烫伤大鼠创面炎性反应与愈合的影响

目的观察海水浸泡对烫伤大鼠创面炎性反应及愈合的影响。方法将144只雄性Wistar大鼠随机分为烫伤对照组和海水浸泡组,每组72只,均造成背部10%TBSA浅Ⅱ度烫伤。海水浸泡组大鼠伤后固定四肢,立即用盛海水的方盆浸泡双前肢以下部分,持续4h；烫伤对照组大鼠则用空方盆模拟浸泡过程。于伤后0(即刻,下同)、6、12、24h采用电解质分析仪测定血清中K~+、Na~+、Cl~-的浓度。于伤前及伤后0、6、12h采用酶联免疫吸附测定法检测血清中肿瘤坏死因子(TNF)α及白细胞介索(IL)6的含量。对两组大鼠创面行大体和组织病理学观察,并记录创面愈合时间。结果海水浸泡组大鼠血清中K~+、Na~+、Cl~-的浓度大多高于烫伤对照组。伤后6h海水浸泡组大鼠血清TNF-α、IL-6含量分别为(140±22)、(160±41)ng/L,均明显高于伤前值(29±15)、(62±17)ng/L及烫伤对照组(120±12)、(124±22)ng/L(P＜0．05)。与烫伤对照组比较,海水浸泡组大鼠创面水肿及局部组织炎性反应加重,创面再上皮化和表皮各层的分化延迟；海水浸泡组创面愈合时间为(16．3±1．6)d,明显迟于烫伤对照组(14．1±1．8)d(P＜0．05)。结论大鼠烫伤后经海水浸泡,可加重创面炎性反应,使创面愈合延迟。     

荷负电气溶胶对大鼠烫伤创面愈合中细胞周期蛋白B1 、细胞周期蛋白C、增殖细胞核抗原表达的影响

目的探讨荷负电气溶胶（aerosols）在烫伤创面愈合过程中对细胞周期蛋白B1（cyclin B1）、细胞周期蛋白C（cyclinC）及增殖细胞核抗原（PCNA）表达的影响。方法建立SD大鼠深Ⅱ度烫伤模型，随机分为荷负电气溶胶组和对照组，每组40只。采用免疫组织化学方法，检测创面愈合过程中cyclin B1、cyclinC及PCNA表达水平。结果荷负电气溶胶组在伤后第2天开始有cyclinC表达于基底细胞胞核，第5天胞核表达显著，第6～10天持续高水平表达。而对照组cyclinC于第3天开始表达于基底细胞胞核，第7天表达显著，第8～10天持续高水平表达。荷负电气溶胶组cyclinC表达明显高于对照组（P〈0．05）。PCNA免疫组化结果显示荷负电气溶胶组和对照组在伤后第3天基底层细胞和毛囊细胞胞核均有表达，荷负电气溶胶组第5天表达显著，而对照组第7天表达显著，提示有大量细胞处于增殖状态。荷负电气溶胶组PCNA表达明显高于对照组（P〈0.01）。而cyclin B1在荷负电气溶胶治疗组和对照组均于伤后第3天有部分表达于基底细胞胞浆和胞核，在第5天胞核表达明显增多，但差异无统计学意义（P〉0．05）。结论荷负电气溶胶照射治疗能有效促进烫伤创面中cyclinC和PCNA的表达，促进细胞增殖，加快创面愈合。     

荷负电气溶胶促进烧伤患者头部取皮区创面愈合效果观察

目的 探讨荷负电气溶胶促进烧伤患者头部取皮区创面愈合的效果.方法 将120例患者随机分为气溶胶组(61例)和对照组(59例),两组均采用电动取皮机取头皮,厚度0.25 mm.取皮后对照组采用常规处理;气溶胶组采用气溶胶对头部取皮区创面进行治疗护理,每次2 h,每天2次,直至创面愈合.结果 两组创面疼痛消失、创面渗出物消失情况及愈合时间比较,差异有显著性意义(P＜0.05、P＜0.01).结论 荷负电气溶胶能有效促进烧伤患者头部取皮区创面愈合.     

荷负电气溶胶治疗大鼠烧伤创面后表皮干细胞的变化

目的探讨荷负电气溶胶对烧伤创面表皮干细胞影响以及其促进创面愈合的机制。方法将清洁级健康Wistar大鼠48只随机分为A、B两组，每个动物背部两侧分别制成实验创面或对照创面，实验创面以荷负电气溶胶照射，A组动物每日照射2次，B组每日照射1次；利用干细胞表达β1整合素的特性，加入整合素β1抗体，通过免疫组织化学染色检测创面表皮干细胞含量的变化，比较各时相点两组创面间的差异。结果A、B两组各时相点实验创面的表皮干细胞数目是同组对照创面的1．5到2倍；而A组实验创面的表皮干细胞数又较B组实验创面增多。结论经用荷负电气溶胶照射大鼠烧伤创面能促进表皮干细胞的增殖。     

类肝素对大鼠深II度烫伤创面愈合的影响

目的 探讨深Ⅱ度烧伤创面早期加深而延迟愈合的机理。方法 采用大鼠深Ⅱ度烫伤模型，创面外用类肝素软膏，观察其对大鼠深Ⅱ度烫伤早期创面病理变化和创面愈合的影响，并测定了烧伤创面含水量，血浆和烧伤创面抗凝血酶Ⅲ（ＡＴ－Ⅲ）、纤维蛋白降解产物（ＦＤＰ）含量、烧伤创面羟脯氨酸含量、Ｉ／Ⅲ型胶原比例、真皮细胞增殖周期及创面愈合时间。结果 创面外用类肝素可以降低烧伤创面水肿程度，增加血浆ＡＴ－Ⅲ活性，增加烧伤创     

Bioactive interleukin-8 is expressed in wounds and enhances wound healing

BACKGROUND: Wound healing is a sequential biological process that involves the integration of chemotaxis of neutrophils, mitosis and migration of keratinocytes, and remodeling of the scar, all of which are regulated by specific soluble mediators. To modulate wound healing specific mediators have to be identified and functionally characterized. Therefore we addressed this study on the polymorphonuclear leukocyte (PMN) attractant interleukin-8 (IL-8) and its function in epidermal wound healing. MATERIALS AND METHODS: Peptide purification, bioassays for PMN chemotaxis, and sequential IL-8 measurements were performed on human wound fluid from burn blisters and skin graft donor sites. Histology for IL-8 immunoreactivity was included. In vitro human keratinocytes were assayed for proliferation, migration, and integrin expression after IL-8 treatment. Wounding experiments with topical IL-8 were performed in a chimeric mouse model. RESULTS: IL-8 was found to be the major bioactive chemoattractant for PMNs in human blister and skin graft donor site wound fluids (mean levels ranging from 173 ng/ml Postoperative Day (POD) 1 to 2130 ng/ml (POD 5)). Released intracellular epidermal IL-8 immunoreactivity at the wound edge was considered as an immediate source of IL-8 while NH(2)-terminal analysis revealed the 77-amino-acid residue form as a second source of IL-8 possibly PMN derived. In vitro experiments on the effect of recombinant human (rh) IL-8 on keratinocyte proliferation revealed a rise in cell number (4.8-fold, ED(50) = 0.6 ng/ml), which was accompanied by an increase in cells in S phase and overexpression of the integrin subunit alpha6. In vivo topically applied IL-8 (1 microg/ml) on human skin grafts in a chimeric mouse model enhanced reepithelialization in IL-8 treated animals over controls due to elevated numbers of mitotic keratinocytes. Wound contraction was significantly diminished by topical IL-8. CONCLUSIONS: These results indicate the sequential function of endogenous IL-8 in all phases of human wound healing. Topical IL-8 may be useful in impaired wound healing.     

Local expression of tumor necrosis factor-alpha and interleukin-10 on wound healing of intestinal anastomosis during endotoxemia in mice

BACKGROUND: This study aimed to evaluate the integrity of anastomotic wound healing after digestive surgery under septic conditions and define the participation of local expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) around the anastomotic segment. MATERIALS AND METHODS: Experimental animals were divided into lipopolysaccharide (LPS) and control groups, which had either LPS or normal saline solution injected into the peritoneal cavity 24 h before transection and anastomosis of the colon. Anastomotic bursting pressure (ABP) and tissue hydroxyproline concentration (HP) were measured as indicators of wound healing. Immunohistochemical staining for TNF-alpha and IL-10 on tissue samples obtained from the anastomotic segment were examined 1, 6, and 24 h after the operation. The reactive cells were counted under light microscopy. RESULTS: ABP and HP were significantly lower in the LPS group than in the control group 7 days after surgery. In the LPS group, TNF-alpha expression increased about threefold over that in the control group 1 h after the operation. TNF-alpha-reactive cells were observed until 24 h after the operation in the LPS group, but not in the control group. On the other hand, IL-10 was not expressed in the control group during the observed period, whereas IL-10 was observed 24 h after the operation in the LPS group. CONCLUSIONS: It is suggested that anastomotic wound healing was impaired after the digestive surgery in animals treated with intraperitoneal LPS, and that local expression of TNF-alpha and IL-10 at the anastomotic site acts as an inhibitory factor in the wound healing process.     

mRNA and protein expression of transcription factor c—fos in burned rats and their effects on wound healing

Objective:To explore the expression of mRNA and its protein in burned rats and their effects of burn wound healing.Methods:A partial-thickness burn of 30% total body surface area was created on the back of 40 Wistar rats.In situ hybridization and immunohistochemical methods were used to exaluate the location and the amount of the c-fos mRNA and its protein in normal skin and the burned skin,respectively,at 3h,6h,1d,3d,7d and 14d after burn.Results:Under a light microscope,both the expression of c-for mRNA and its protein could be found in the normal skin,but their induction levels were much higher in the burned skin.The level of for protein expression reached peak at 3h after burn while that of c-for mRNA reached peak at 6h after burn.Conclusions:The expression of c-fos can be induced by burns.And the peak level expression of c-for mRNA comes later than that of c-fos protein.It indicates that the action of fos protein is induced by post-translational modification of pre-existing fos molecules.     

Influence of oxygen on wound healing

Oxygen has an important role in normal wound healing. This article reviews the evidence concerning the role of oxygen in wound healing and its influence on the different stages of wound healing. The evidence reviewed has demonstrated that improving oxygenation may be helpful in limiting wound infection, although there is a lack of good quality studies on the role of oxygen in the proliferative phase and in reepithelialisation. Overall, the relationship between oxygen and wound healing is complex. Knowledge of this aspect is important as many treatment modalities for refractory wounds are based on these principles.     

银锌霜在大面积烧伤中的应用

目的总结1991年1月至1995年11月银锌霜在48例TBSA大于30％的烧伤病人创面的应用。方法将同期应用碘络醚的35例病人作为对照,两组病人的平均年龄、烧伤面积、Ⅲ度面积无显著差别,用药方式均以半暴露为主。结果银锌霜组能显著增加细菌转阴率,减少抗生素应用时间及植皮手术次数,缩短愈合时间。结论银锌霜具有较强的抗感染能力,是大面积烧伤病人的良好外用药。     

荷负电气溶胶对大鼠烫伤创面愈合中细胞周期蛋白B1 、细胞周期蛋白C、增殖细胞核抗原表达的影响

目的探讨荷负电气溶胶(aerosols)在烫伤创面愈合过程中对细胞周期蛋白B1(cyclin B1)、细胞周期蛋白C(cyclin C)及增殖细胞核抗原(PCNA)表达的影响.方法建立SD大鼠深Ⅱ度烫伤模型,随机分为荷负电气溶胶组和对照组,每组40只.采用免疫组织化学方法,检测创面愈合过程中cyclin B1、cyclin C及PCNA表达水平.结果荷负电气溶胶组在伤后第2天开始有cyclin C表达于基底细胞胞核,第5天胞核表达显著,第6～10天持续高水平表达.而对照组cyclin C于第3天开始表达于基底细胞胞核,第7天表达显著,第8～10天持续高水平表达.荷负电气溶胶组cyclin C表达明显高于对照组(P<0.05).PCNA免疫组化结果显示荷负电气溶胶组和对照组在伤后第3天基底层细胞和毛囊细胞胞核均有表达,荷负电气溶胶组第5天表达显著,而对照组第7天表达显著,提示有大量细胞处于增殖状态.荷负电气溶胶组PCNA表达明显高于对照组(P<0.01).而cyclin B1在荷负电气溶胶治疗组和对照组均于伤后第3天有部分表达于基底细胞胞浆和胞核,在第5天胞核表达明显增多,但差异无统计学意义(P>0.05).结论荷负电气溶胶照射治疗能有效促进烫伤创面中cyclin C和PCNA的表达,促进细胞增殖,加快创面愈合.     

The effect of interleukin-2 administration on wound healing in adriamycin-treated rats

Adriamycin (doxorubicin hydrochloride), an effective chemotherapeutic drug, is also a potent inhibitor of wound healing. Conversely, certain polypeptide growth factors are capable of stimulating fibroblasts to secrete collagen, thus enhancing wound healing. The purpose of this study was to determine if interleukin-2 (IL-2), a T-cell growth factor, could reverse the wound healing deficit caused by Adriamycin. Adriamycin treatment caused a significant decrease in wound-breaking strength (P less than 0.005). IL-2 administration increased wound-breaking strength in Adriamycin-treated animals (2126 g vs 1549 g, P less than 0.005). In control animals, IL-2 did not increase wound-breaking strength significantly (2708 g vs 2608 g, P greater than 0.1). Histologically, wounds from Adriamycin-treated animals were less cellular, demonstrated less collagen in the dermis, and a lesser degree of capillary ingrowth. The number of fibroblasts in the dermal layer was increased in animals receiving IL-2. Control rats gained an average of 1.4% of their original body weight, while Adriamycin-treated rats lost an average of 19% of their original body weight (P less than 0.0005). IL-2 administration did not influence weight loss or gain. Hematologically, animals receiving Adriamycin had lower hemoglobin and hematocrit values and higher platelet counts. There were no differences in total white blood cell counts; however, animals receiving Adriamycin showed a predominance of polymorphonuclear leukocytes and a relative decrease in lymphocytes. Animals receiving IL-2 demonstrated a significant eosinophilia. (1) Adriamycin impairs normal wound healing. (2) Interleukin-2 administration improves the wound healing impairment caused by Adriamycin. (3) Interleukin-2 appears to increase infiltration of inflammatory cells, fibroblasts, and capillaries into the wound, which may account for the observed increase in wound breaking strength.