Immunohistochemical analysis of markers for different macrophage phenotypes and their use for a forensic wound age estimation

A total of 117 vital skin wounds (post infliction intervals between a few seconds and 7 months), 20 postmortem wounds and skin specimens with beginning or advanced signs of putrefaction were investigated. Different markers for macrophage maturation (27 E 10, RM 3/1, 25 F 9, G 16/1) were analyzed by immunohistochemistry. The early stage inflammation marker 27 E 10 stained macrophages, but also monocytes and neutrophilic granulocytes localized in blood vessels or bleeding induced postmortem and therefore provided no further information for a forensic wound age estimation in comparison to the routine histological detection of macrophages. The antigens recognized by the RM 3/1- (intermediate stage inflammation marker) and 25 F 9-antibodies (late stage inflammation marker) were expressed exclusively by histiocytes and inflammatory cells that had migrated from the blood vessels as part of the acute inflammatory response associated with an intravital reaction. The morphometrical analysis revealed positive results (defined as at least a two-fold increase in number in 2 or more microscope fields when compared to the maximum value of histiocytes found in uninjured skin) for the RM 3/1- or 25 F 9-antibody earliest in wounds aged 7 or 11 days, respectively. Similarly to the 25 F 9-antibody, the chronic stage inflammation marker (G 16/1) reacted with a macrophage subpopulation first detectable 12 days after wounding but showed positive results in a comparably reduced percentage of cases. On the other hand, this marker did not stain a relevant number of resident macrophages thus facilitating the evaluation of the specimens. The markers 27 E 10, RM 3/1 and 25 F 9 are also useful for the evaluation of slightly - even though the staining intensity was considerably reduced - but not advanced putrefied skin. Therefore, the immunohistochemical analysis of the corresponding antigens can possibly contribute to an age estimation of wounds with advanced post infliction intervals obtained from corpses with longer - but limited - postmortem intervals.

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Zusammenfassung Insgesamt wurden 117 vitale Hautwunden (Überlebenszeit wenige Sekunden bis 7 Monate), 20 postmortal gesetzte Verletzungen sowie Haut mit leichten bzw. fortgeschrittenen Fäulnisveränderungen untersucht und verschiedene Marker der Makrophagen-Differenzierung (27 E 10, RM 3/l, 25 F 9 und G 16/1) analysiert. Der early stage inflammation marker 27 E 10 färbte neben Makrophagen auch Monozyten und neutrophile Granulozyten, die innerhalb von Blutgefäßen bzw. in postmortal gesetzten Blutungen lokalisiert waren und liefert somit keine Informationen zum Wundalter, die über die Möglichkeiten des Routine-histologischen Nachweises von Makrophagen hinausgingen. Die von den Antikörpern RM 3/1 (intermediate stage inflammation marker) und 25 F 9 (late stage inflammation marker) erkannten Antigene wurden ausschließlich von Histiozyten und reaktiv eingewanderten Makrophagen exprimiert. Die morphometrische Analyse ergab positive Ergebnisse (definiert als ein mindestens zweifacher Anstieg der Zellzahl in zwei oder mehr Gesichtsfeldern verglichen mit der maximal feststellbaren Zahl an Histiozyten in unverletzter Haut) bei Verwendung der Antikörper RM 3/1 bzw. 25 F 9 frühestens 7 bzw. 11 Tage nach Wundsetzung. Ab 12 Tagen Wundalter reagierte der chronic stage inflammation marker G 16/1 erstmals positiv. Das Antigen ließ sich insgesamt allerdings in einem geringeren Prozentsatz der untersuchten Wunden darstellen. Vorteilhaft ist jedoch das Fehlen einer relevanten Expression durch Histiozyten, wodurch die Auswertung der Präparate erleichtert wird. Die entsprechenden Antigene lassen sich zudem in leicht - wenn auch in einer deutlich geringeren Färbeintensität -, aber nicht forgeschritten fäulnisveränderter Haut nachweisen, so daß deren immunhistochemische Darstellung gegebensfalls auch zur Beurteilung von länger überlebten Verletzungen an Leichen mit etwas fortgeschrittener Liegezeit herangezogen werden kann.

     

Forensic application of VEGF expression to skin wound age determination

An immunohistochemical study combined with morphometry was carried out to examine the time-dependent expression of vascular endothelial growth factor (VEGF) using 53 human skin wounds with different wound ages (groups I: 0–12 h, II: 1–4 days, III: 7–14 days and IV: 17–21 days). In the human wound specimens aged 4–12 h, neutrophils recruited at the wound showed no positive signals for VEGF. With an increase in wound ages of 7 days, granulation tissue and angiogenesis were observed, with the migration of macrophages and fibroblasts of which the cytoplasm expressed VEGF-positive reactions. Morphometrically, the average VEGF-positive ratio was highest in group III, followed by that of group IV. In groups III and IV, 13 out of 26 wound samples had VEGF-positive ratios of more than 50%. However, all of the wound samples in groups I and II showed VEGF-positive ratios of less than 50%. With regard to the practical applicability and forensic validity, these observations suggest that a VEGF-positive ratio of more than 50% possibly indicates a wound age of 7 days or more.     

Estimation of wound age and VCAM-1 in human skin

To estimate the age of skin wounds, the endothelial adhesion molecule VCAM-1 (CD 106) was detected in paraffin sections after autoclaving and using the ABC technique. The percentage of VCAM-1 positive blood vessels was determined after the blood vessels had been marked with PECAM-1 (CD 31). Low positive staining reactions were observed for VCAM-1 on endothelial cells of uninjured skin in 18% of the samples. In injured skin, 51% of the cases investigated showed a VCAM-1 expression. Strong positive staining reactions were observed 3 h at the earliest and 3.5 days at the latest after the time of injury. The immunohistochemical results for VCAM-1 differed significantly between the injured and uninjured skin (P < 0.01). In a few cases VCAM-1 was detected (n = 6) at low intensity in postmortem skin wounds and a moderate to strong expression of VCAM-1 is indicative of the vitality of the wound. The detection of VCAM-1 can be used for estimating the age of wounds in forensic applications if the degree of expression of further adhesion molecules, especially that of selectins, is taken into account. Received: 8 April 1998 / Received in revised form: 15 July 1998     

Detection of fibrocytes in human skin wounds and its application for wound age determination

Fibrocytes, a newly identified cell type, are bone marrow-derived mesenchymal progenitors that coexpress hematopoietic cell antigens and fibroblast products. In this study, a double-color immunofluorescence analysis was carried out using anti-CD45 and anti-collagen type I antibodies to examine the time-dependent appearance of fibrocytes, using 53 human skin wounds with different wound ages (group I, 0–3 days; group II, 4–7 days; group III, 9–14 days; and group IV, 17–21 days). In wound specimens with an age of less than 3 days, CD45⁺/collagen type I⁺ fibrocytes were not detected. The fibrocytes were initially observed in wounds aged 4 days, and their number increased in lesions with advances in wound age. In a semiquantitative morphometrical analysis, the average number of fibrocytes was highest in the wounds of group III. These findings imply that human skin wounds containing fibrocytes are at least 4 days old. Moreover, a fibrocyte number of over 10 indicates a wound age between 9 and 14 days (i.e., group III). Based on the average number of fibrocytes in each group, a fibrocyte number of over 15 more strongly suggests a wound age of 9–14 days. Together, our observations indicate the participation of fibrocytes in wound healing of human skin inducing the accumulation of extracellular matrix components, and therefore, detection of fibrocytes could be a useful marker for wound age determination.     

Expression of oxygen-regulated protein 150 (ORP150) in skin wound healing and its application for wound age determination

Immunohistochemical study combined with morphometry was carried out to examine the expression of oxygen-regulated protein 150 (ORP150) using 58 human skin wounds of different ages (group I, 0-12 h; II, 1-5 days; III, 7-14 days; and IV, 17-21 days). In human wound specimens aged 4-12 h, neutrophils recruited at the wound showed no positive signals for ORP150. With the increase in wound age of >/=7 days, granulation tissue and angiogenesis were observed, with the migration of macrophages and fibroblasts with ORP150-positive reactions. In semi-quantitative analysis, the average of ORP150-positive ratios in group III was highest. In group III, all samples had an ORP150-positive ratio of >40%, and 17 samples showed >50%. In group IV, three out of ten samples showed a positive ratio of 40-45% and the remaining seven cases less than 40%. Collectively, with regard to the practical applicability with forensic safety, these observations suggest that an ORP150-positive ratio of >50% strongly indicates a wound age of 7-14 days.     

Immunohistochemical detection of chemokines in human skin wounds and its application to wound age determination

Immunohistochemical studies on the time-dependent expression of the chemokines such as interleukin (IL)-8, monocyte chemoattractant protein (MCP)-1 and macrophage inflammatory protein (MIP)-1α were performed on 50 human skin wounds with different wound ages (group I 0–12 h, group II 1–4 days, group III 7–14 days and group IV 17–21 days). In the wound specimens with wound ages between 4 and 12 h, neutrophils mainly showed positive reactions for IL-8, MCP-1 and MIP-1α. With increasing wound ages, macrophages and fibroblasts were positively stained with anti-IL-8, MIP-1α and MCP-1 antibodies. Morphometrically, there was a similar distribution in the positive ratios of the inflammatory cells among IL-8, MCP-1 and MIP-1α. The positive ratios of each chemokine were very low in group I and a considerable increase of the positive ratios in each chemokine was observed in group II (mean ± standard error IL-8: 59.8 ± 2.1%, MCP-1: 42.4 ± 3.1% and MIP-1α: 50.4 ± 3.7%). Although the positive ratios for each chemokine gradually decreased according to the wound age, the mean positive ratios in groups III and IV were significantly higher than those in group I. From the forensic aspect, these chemokines are considered useful markers for wound age determination. Thus, ratios of > 50% for IL-8, > 30% for MCP-1 or > 40% for MIP-1α indicate a wound age of at least 1 day. Moreover, the combined investigation of these three chemokines can make wound age determination more objective and accurate. Received: 30 May 2001 / Accepted: 14 August 2001     

Temporal expression of wound healing-related genes in skin burn injury

Determination of the age of burns, as well as of wounds induced mechanically, is essential in forensic practice, particularly in cases of suspected child abuse. Here, we investigated temporal changes in the expression of 13 genes during wound healing after a burn. The expression of cytokines (IL-1β, IL-6, IL-10, TNF-α, and IFN-γ), chemokines (KC, MCP-1), proliferative factors (TGF-β, VEGF), proteases (MMP-2, 9, 13) and type I collagen in murine skin was examined by real-time PCR at 3, 6, 9, and 12 h and 1, 2, 3, 5, 7, and 14 days after a burn. Based on macroscopic and histological appearance, the healing process of a burn consists of 3 phases: inflammatory (from 3 h to 1 day after the burn), proliferative (from 1 to 7 days), and maturation (from 7 to 14 days). Expression of IL-1β, IL-6, TNF-α, IFN-γ and KC increased significantly in a biphasic pattern from 3 or 6 h to 12 h or 1 day and from 3 or 5 days to 7 days. Expression of MCP-1 increased significantly from 6 h to 5 days. Expression of both IL-10 and TGF-β increased significantly from 12 h to 7 days. Expression of VEGF, MMP-2, MMP-13 and type I collagen increased significantly from 3 days to 7 or 14 days. Expression of MMP-9 increased significantly from 6 h to 14 days. Our results suggest that evaluating the expression of a combination of these genes would enable the exact estimation of the age of a burn.     

Bioinformatics analysis of sequential gene expression profiling after skin and skeletal muscle wound in mice

It is of great value to use bioinformatics methods to screen the core differentially expressed genes (DEGs) at different times after mouse skin and skeletal muscle wound, and to explore the relationship between them and the wound age. To this end, we downloaded the gene expression profiles of GSE140517 and GSE23006 from the NCBI-GEO gene database, used GEO2R online tools and Venn diagrams to screen out DEGs at different times and common-DEGs. The Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) channel analysis were carried out through the DAVID website respectively. Use STRING tool to build a Protein-protein Interaction (PPI) network, and use Cytoscape software to screen out core DEGs. The results showed that 13, 53, 43 and 13 core DEGs were screened out in the 6 h, 12 h, 24 h and common-DEGs group after wound. There were 7 core DEGs (Cxcl2, Cxcl3, Il1b, Ptgs2, Cxcl1, Timp1, Ccl3) in both the different time point and the common DEGs group. Meanwhile, there are 1 core DEGs (Ccl4) specifically expressed in the 6 h, 29 specifically expressed core DEGs (Isg20, Rtp4, Fcgr1, Ifi44, Trim30a, etc.) in the 12 h, and 18 specifically expressed core DEGs (Ccr7, Myd88, Igsf6, Ccr2, Gpsm3, etc.) in the 24 h, there are 6 core DEGs (Ccl4, Ccl7, Saa3, Cxcl5, Ccl2, Lcn2) specifically expressed in the common-DEGs group. The results of GO and KEGG analysis showed that the deterioration and exudation of the inflammatory response were the main cause at 6 h after wound. In addition to inflammation at 12 h and 24 h, the systemic immune response against viral and bacterial infections also gradually increased. In summary, the core DEGs selected in this study have combined characteristics, consistent with the healing function at the corresponding time point, and they are also has specificity and correlation with wound age. Therefore, by detecting the changes in the expression of co-expressed core DEGs at different times after wound, as well as detecting specific expressed DEGs at a specific time point or a specific period of time, it is very promising to provide help for the wound age estimation. However, limited by the GSE140517 gene expression profile in the database, only the difference in gene expression at different times within 24 h after wound was explored, and the research on the late wound age still needs to be further in-depth.     

大鼠急性肺损伤病理变化的形态计量研究

目的 用体视学方法定量研究大鼠经胸部撞击复合内毒素所致急性肺损伤(acute lung injury,ALI)模型的组织病理学特点及变化规律. 方法 12只SD大鼠按随机数字表法分为正常对照组、ALI致伤1,2,7d组,每组3只.各组取肺组织行HE染色,分别测试肺实质百分比、肺泡间隔厚度、肺泡截面积及肺泡腔内炎性细胞计数. 结果 ALI致伤1,2d组肺泡腔内炎性细胞计数分别为(13.08 ±6.60)个、(26.81±16.86)个,正常对照组为(0.89±0.78)个,致伤组均较对照组明显增加(P＜0.01).ALI致伤1,2,7d组肺实质百分比分别为(23.77±3.91)％、(27.32±5.95)％、(44.57±6.46)％,正常对照组为(19.44±2.48)％,致伤组均较对照组明显增加(P＜0.01).ALI致伤1,2,7d组肺泡间隔分别为(3.50±2.18) μm、(7.67±7.14) μm、(17.23±13.08) μm,正常对照组为(2.87±1.75) μm,致伤组均较对照组逐渐增厚(P＜0.05).ALI致伤1,2,7d组肺泡截面积分别为(1 653.90±950.77)μm2、(1 328.15±802.30) μm2、(1 055.20±1 002.75) μm2,正常对照组为(1 641.30 ±906.60)μm2,致伤组均较对照组逐渐减少(P＜ 0.05). 结论 大鼠经胸部撞击后复合静脉注射内毒素可建立病理形态上符合ALI的动物模型.通过体视学方法定量检测可反映ALI的组织病理学特点及变化规律.     

Immunohistochemical localization of fibronectin as a tool for the age determination of human skin wounds

Summary We analyzed the distribution of fibronectin in routinely embedded tissue specimens from 53 skin wounds and 6 postmortem wounds. In postmortem wounds a faint but focal positive staining was exclusively found at the margin of the specimens which dit not extend into the adjacent stroma. Vital wounds were classified into 3 groups. The first comprising lesions with wound ages ranging from a few seconds to 30 min, the second comprising those with wound ages upt to 3 weeks, and the third group with lesions more than 3 weeks old. Ten out of 17 lesions with a wound age up to 30 min showed a clear positive reaction within the wound area. Three specimens in this group were completely negative, while in 4 additional cases the result was not significantly different from postmortem lesions. These 7 cases were characterized by acute death with extremely short survival times (only seconds). In wounds up to 3 weeks old fibronectin formed a distinct network containing an increasing number of inflammatory cells corresponding to the wound age. In 2 cases with a survival time of 17 days and in all wounds older than 3 weeks fibronectin was restricted to the surface of fibroblasts and to parallel arranged fibers in the granulation tissue without any network structures. We present evidence that fibronectin is a useful marker for vital wounds with a survival time of more than a few minutes. Fibronectin appears before neutrophilic granulocytes migrate into the wound area. Since a faint positive fibronectin staining is seen in postmortem lesions and bleedings, we propose that only those wounds which show strong positive fibronectin staining also extending into the adjacent stroma should be regarded as vital.This study was supported by a grant from the Deutsche Forschungsgemeinschaft (grant Ei 209/3-1) and by a grant from the Friedrich-Baur-Stiftung University of Munich     

Simultaneous detection of eight cytokines in human dermal wounds with a multiplex bead-based immunoassay for wound age estimation

We performed quantification of IL 2, IL 4, IL 6, IL 8, IL 10, GM-CSF, IFN γ, and TNF α in human dermal wounds for wound age estimation. The proliferation of dermal cells and infiltration of inflammatory cells were also analyzed. Neutrophils and macrophages were detected from 2 h post-injury, and strong infiltrations were seen at 33–49 h. T and B lymphocytes also infiltrated simultaneously from 71 h. Strong proliferation of fibroblasts were shown from 246 h, and thickening of the epidermis from 71 h. IL 10, GM-CSF, IFNγ, and TNF α increased from the early phase of dermal wound healing, IL 6 exclusively in the middle phase, IL 2, IL 4, and IL 8 from the middle phase to the late phase. Among the cytokines analyzed in the present study, IL 6, IL 8, IFNγ, and TNF α were strongly expressed. Results of the present study suggest that multiplex cytokine analysis at the wound site can be useful for wound age estimation. In addition, multiplex data obtained from the same sample with a single method would demonstrate more accurate interactions of cytokines during dermal wound healing. Although the present study was oriented to practical forensic pathology, the data obtained would be informative for various fields of medicine.     

Histological and enzyme histochemical parameters for the age estimation of human skin wounds

Routine histological staining techniques form the basis of a forensic age estimation of human skin wounds and the determination of vitality is aided by the detection of neutrophilic granulocytes which appear earliest about 20–30 min after wounding. A clear granulocyte infiltration and a significant increase in the number of macrophages indicates a post infliction interval of at least several hours. Macrophages containing incorporated particles such as lipophages, erythrophages or siderophages appear earliest at a wound age of 2–3 days similarly to extracellular deposits of hemosiderin, whereas the rarely detectable iron-free pigment hematoidin and spot-like lymphocytic infiltrates in the granulation tissue appear approximately one week or more after wounding. A complete reepithelialization of surgically treated and primarily healing human skin lesions can be expected earliest 5 days after wound infliction and the absence of a complete new epidermal layer indicates a survival time of less than 21 days. Enzyme histochemical methods allow a wound age differentiation especially in the range of a few hours. An increase in nonspecific esterases can be observed earliest approximately 1 hour after wounding followed by other enzymes such as acid phosphatase ( 2 h), ATPase ( 4 h), aminopeptidase ( 4 h) or alkaline phosphatase ( 4 h). Positive results, however, cannot be regularly found. Therefore, the detection of reactive changes is useful for a wound age estimation whereas negative findings, which in general must be interpreted with caution, can provide information only in a limited number of histological parameters.Dedicated to Prof. Dr. W. Eisenmenger on the occasion of his 50th birthday     

Radial alveolar count as a tool for the estimation of fetal age

A total of 53 normal fetuses with a gestational age ranging from 15 up to 39 weeks was investigated and the radial alveolar count (RAC) was estimated as a parameter for lung maturation. Values lower than 2.0 could only be found in lungs of fetuses aged less than 18 weeks. Between 18 and 25 weeks of gestation, relatively constant levels of RAC were observed but with considerable interindividual variation. In fetuses with a gestational age of more than 25 and especially 30 weeks, a slight or rapid increase in RAC occurred respectively. Values lower than 3.0 were found up to a fetal age of less than 30 weeks and a RAC of more than 4.0 was only found in lungs of fetuses aged more than 30 weeks. Values exceeding 6.0 occurred only in fetuses at near full-term birth. Since the estimation of RAC overcomes the effects of varying degrees of alveolar collapse, such an analysis also seems to be useful for the determination of fetal age in cases of advanced putrefaction. Received: 9 August 1996 / Received in revised form: 22 November 1996     

Studies on mRNA expression of tissue-type plasminogen activator in bruises for wound age estimation

We investigated mRNA expression of tissue-type plasminogen activator (tPA) and inflammatory cell dynamics for wound age estimation of bruises in mice. Neutrophils were detected from 1 h post-injury. Up to 8 h, they accumulated in subcutaneous tissue and the lower part of the dermis, and thereafter they extended to all the layers. Macrophages became detectable 3 h post-injury, and moderate infiltration of lymphocytes was seen from 144 h. In addition, epidermal thickening was also seen from 72 h. tPA mRNA expression peaked at 1 h, and increased slightly at 72 h post-injury. tPA mRNA was detected in epidermal cells, fibroblasts, and endothelial cells before and after injury, from 3 h in neutrophils and from 72 h in macrophages, respectively. This study presents the time-dependent expression of tPA mRNA in bruises in relation to temporal histologic characteristics during wound healing, which was considered to be useful for wound age estimation. Furthermore, it is suggested that tPA plays an important role in the first step of tissue remodeling.     

Deferiprone reduces hemosiderin deposits in the brain of a patient with superficial siderosis

A man with superficial siderosis showed improvement in symptoms and reduction in hemosiderin by MR imaging following treatment with deferiprone, a lipid-soluble iron chelator.     

海水浸泡对伤口愈合时间影响的动物实验观察

目的 观察创伤合并海水浸泡对兔伤口愈合时间的影响.方法 以家兔为实验动物,建立背部双侧圆形创伤模型,伤后随机分为对照组和实验组,每组8只,对照组不浸泡,实验组伤后置海水中浸泡20 min.观察2组创面肉芽组织的生长状况,采用直尺测量法及照相法记录各组伤口大小的变化及最终愈合时间.结果 对照组伤口愈合时间为12～13 d,实验组伤口愈合时间为16～18 d.对照组的肉芽组织形成时间早于实验组.结论 创伤后合并海水浸泡可导致伤口愈合时间明显延长.     

国内负压创面治疗技术临床研究现状的循证分析

目的 采用循证医学的方法,对国内负压创面治疗技术的临床现状进行系统性研究. 方法 系统性检索2008年12月前国内发表的关于负压创面治疗技术的文献,并对结果进行证据分级和质量等级的评估. 结果 检索到符合标准的文献5篇(均为随机对照试验):1篇证据分级为1b,质量等级为"优",余4篇证据分级为2b,质量等级为"差". 结论 对于负压技术相对宽泛的适应证,目前国内缺乏足够的证据证明其疗效.今后需进一步规范该技术的科学研究及临床应用,包括注重选题和立题的创新性、试验设计的完善性和实用性、治疗过程的标准化等问题.提高发表文献的质量,以更好地指导临床工作.     

The dynamics of inflammatory cytokines in the healing process of mouse skin wound: A preliminary study for possible wound age determination

The dynamics of inflammatory cytokines such as interleukin-1 (IL-l, interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF) during the healing process of mouse skin wounds were examined using the enzyme-linked immunosorbent assay and immunostaining. The applicability of this examination for wound age estimation is discussed from the perspective of forensic pathology. After wound induction, mice were sacrificed at intervals ranging from 0 to 240 h. The levels of TNF and IL-1 began to elevate rapidly after wounding and reached a peak at 3 h. The IL-l level reached a peak at 6 h, and IL-6 peaked at 12 h. An infiltration of numerous leukocytes, indicatingacute inflammation, was observed at 3 and 6 h, and the main source of the cytokines was immunohistochemically identified as neutrophils. These results indicate that TNF and IL-1 play an important role in the commencement of inflammation. Rebound of cytokine levels, i.e. a re-increase, was observed at 72 h after wounding. Histological examination of the 72-h-old wound showed migration of fibroblasts and the formation of new granulation tissues, indicating the proliferative stage of the wound healing process. These experimental findings indicate that these cytokines have a close relationship to wound remodeling as well as to inflammation. From the viewpoint of forensic pathology, it is considered that inflammatory cytokines may become one of the markers for wound age estimation, but further studies are needed, especially those involving the investigation using human wound specimens with known time intervals after injury.This study was presented at the 79th Congress of the Medico-Legal Society of Japan (Yamagata, May 1995), the 16th Annual Meeting of the Japanese Society of Inflammation (Tokyo, July 1995) and the 74th Annual Meeting of the German Society of Legal Medicine (Aachen, September 1995).     

精氨酸对糖尿病大鼠伤口愈合的影响

目的 观察精氨酸对糖尿病大鼠伤口愈合的影响。方法 成年雄性Lewis大鼠，随机分为糖尿病组及正常对照组，前者以链脲佐菌素复制糖尿病模型，后者注射等鼙枸橼酸钠缓冲液；7d后住大鼠背部制作伤口，置入聚乙烯乙醇海绵，上述两组大鼠再随机分为精氨酸治疗组及盐水治疗组，每组10只大鼠；精氨酸治疗组每天腹腔注射L-精氨酸1g/kg，盐水治疗组每天注射等量等渗盐水；10d后大鼠处死取样，观察其伤口抗断张力、羟脯氨酸含量、前胶原mRNA表达。结果 与正常大鼠比较，糖尿病大鼠伤口抗断张力、羟脯氨酸含量、前胶原mRNA表达显著降低；与等渗盐水治疗组比较，精氨酸治疗组大鼠伤口抗断张力、羟脯氨酸含量、前胶原mRNA表达显著提高。结论 精氨酸可以有效促进糖尿病动物的伤口愈合。