HSP47在胎鼠皮肤创口愈合中的表达

胚胎时期唇腭裂修复手术无瘢痕愈合的实验研究已有很多报道，学者们建立了许多动物模型来研究胎儿皮肤创口愈合的过程。结果发现胎儿和成人皮肤创口的愈合过程存在显著差异。胎儿皮肤创口具有愈合速度快、无炎症反应、透明质酸含量较高等特点。     

Intra-uterine healing of fetal rat oral mucosal, skin and cartilage wounds

Orofacial wounds, including rupture of the palate, were surgically created in utero on 19½ day Sprague Dawley rat fetuses. Six hours prior to sacrifice 0.1 mgm/100 gm maternal body weight of colchicine was injected subcutaneously. Five fetuses in each group of operated and unoperated controls were sacrificed at 0, 24, 48 and 72 h postoperatively. Serial histological sections in the frontal plane were stained with hemotoxylin and eosin and modified Mallory's. Rapid and complete epithelialization of oral and skin wounds was observed within the 72 h period. There was a complete lack of inflammatory response, scab formation and involvement of nasal epithelium in healing. Nasal cartilage wounds rapidly rounded off and the perichondrium reformed without evidence of callus formation. These findings are discussed in regard to the etiology of some congenital defects and the prospect of fetal surgical correction of orofacial deformity.     

Expression of heat shock protein 47 in the periodontal ligament during orthodontic tooth movement

The aim of this study was to investigate the kinetics of heat shock protein 47 (HSP47) and proliferating cell nuclear antigen (PCNA) immunohistochemistry in periodontal ligament (PDL) cells during orthodontic tooth movement in a mouse model. An orthodontic appliance was set between the upper incisors and the upper left first molar. The mice were killed 2, 6 and 10 days after initiation of orthodontic tooth movement. Computer-assisted image analysis was used to compare the quantitative expression of HSP47 in the PDL. HSP47 expression was significantly higher on the tension side 2 days after application of the appliance, whereas no significant change was observed on the pressure side at any time point. Furthermore, the PCNA labelling indices of PDL cells were increased significantly on the tension side 6 and 10 days after application of the appliance, and on the pressure side 2, 6 and 10 days after application of the appliance. These data suggest that collagen is metabolised predominantly on the tension side, and that PDL cells actively proliferate on both the tension and pressure sides during orthodontic tooth movement.     

Intra-uterine healing of fetal rat oral mucosal, skin and cartilage wounds.

Orofacial wounds, including rupture of the palate, were surgically created in utero on 19 1/2 day Sprague Dawley rat fetuses. Six hours prior to sacrifice 0.1 mgm/100 gm maternal body weight of colchicine was injected subcutaneously. Five fetuses in each group of operated and unoperated controls were sacrificed at 0, 24, 48 and 72 h postoperatively. Serial histological sections in the frontal plane were stained with hemotoxylin and eosin and modified Mallory's. Rapid and complete epithelialization of oral and skin wounds was observed within the 72 h period. There was a complete lack of inflammatory response, scab formation and involvement of nasal epithelium in healing. Nasal cartilage wounds rapidly rounded off and the perichondrium reformed without evidence of callus formation. These findings are discussed in regard to the etiology of some congenital defects and the prospects of fetal surgical correction of orofacial deformity.     

Antisense oligonucleotide against 47-kDa heat shock protein (Hsp47) inhibits wound-induced enhancement of collagen production

It is well known that excessive collagen synthesis during the wound-healing process causes scar formation. Our recent in-vivo study indicates that antisense treatment against 47-kDa heat shock protein (Hsp47), a collagen-specific molecular chaperone, relieves scar formation following skin wounds in rats [Wang et al., Plast. Reconstr. Surg., in press]. In order to understand the mechanism of this phenomenon, we examined the effects of antisense treatment on the expression of mRNAs and proteins of Hsp47 and collagens in fibroblasts derived from wounded rat tongues. Hsp47 and procollagen alpha1(I) and alpha1(III) mRNAs were consistently increased after wounding and were maximal at day 5 post-injury. Treatment with antisense oligonucleotide against Hsp47 efficiently blocked the production of procollagen alpha2(I) and alpha1(III) proteins, but had little effect on their mRNA levels. Therefore, we conclude that antisense oligonucleotide against Hsp47 inhibits the production of procollagen type I and III proteins in fibroblasts derived from wounded tongues, overcoming the increase in their mRNAs.     

Immunolocalization of heat shock proteins 27 and 47 during repair of induced oral ulcers

Heat shock proteins (Hsps) 27 and 47 are involved in the control of apoptosis, cell migration, and collagen synthesis. There is some understanding of the immunolocalization of these proteins during the repair process in skin and gastrointestinal mucosa, but their expressions in normal and injured oral mucosa are unknown. The aim of this study was to analyze the immunolocalization and intensity of these proteins in oral ulcers induced in rats and to compare these expression levels with those reported in skin and gastric mucosa. Ulcers were induced on the ventral surface of the tongues of rats. The rats were then euthanized at 0, 24, 48, 72, and 120 h. Hsp27 expression remained low in the first hours of repair, but was higher at 72 h, mainly in the migrating epithelium. Expression of Hsp47 was high at 48 h, mainly in fibroblasts, cells of the vascular wall, and basal keratinocytes of migrating epithelium. In the control group, expressions of these proteins were low, which indicates that these Hsps are constitutive proteins in oral mucosa. Expression levels were similar to those reported in the healing of skin lesions and gastric ulcer, suggesting a common mechanism of Hsp activation in the repair of these tissues.     

热休克蛋白60、70在口腔扁平苔藓中表达的研究

目的　探讨热休克蛋白 (heatshockprotein ,HSP) 60和HSP70在口腔扁平苔藓病变中的作用。方法　对 62例口腔扁平苔藓、1 0例正常口腔粘膜、2 1例慢性盘状红斑狼疮、1 0例粘膜良性淋巴组织增生病 ,46例白斑进行免疫组织化学SP法染色 ,分析HSP60、HSP70在口腔扁平苔藓中的表达 ;对 1 2例口腔扁平苔藓和 5例正常粘膜进行逆转录PCR实验 ,观察HSP60、HSP70mRNA的变化。结果　HSP60在口腔扁平苔藓病损区的表达较其他各组显著增强 ,差异有显著性 (P <0 0 0 1 )。HSP70在糜烂型口腔扁平苔藓病损区的表达下调。RT PCR结果显示 ,HSP60、HSP70mRNA表达增强。结论　HSP60及HSP70在口腔扁平苔藓的发病中起重要作用     

热休克蛋白27在舌白斑中的表达及意义

目的探讨热休克蛋白（HSP）27在舌白斑中的表达及意义。方法对19例正常舌黏膜、31例舌白斑的石蜡切片用免疫组化法检测HSP27的表达,并对其表达水平进行统计学分析。结果在正常舌黏膜上皮中,HSP27主要位于棘层,呈棕黄色或棕褐色颗粒,在胞浆中表达;舌白斑HSP27分布于上皮的全层。与正常舌黏膜比较,舌白斑HSP27表达水平明显升高（P〈0．001）;不吸烟患者HSP27的表达水平明显高于吸烟患者（P〈0．05）;不同年龄或不同性别患者HSP27的表达水平间差异无统计学意义（P〉0．05）。结论HSP27表达的改变与舌白斑的发生有关。     

雌激素对大鼠咬肌热休克蛋白70表达的影响

目的探讨雌激素对大鼠咬肌热休克蛋白（HSP）70表达的影响。方法将60只雌性未孕12周龄Sprague-Dawley大鼠随机分成3组,每组20只。假手术组大鼠在轻微骚动卵巢后缝合,去卵巢组大鼠去除卵巢后缝合,雌激素替代组大鼠去卵巢后接受雌二醇替代治疗。在实验的第4周和8周,每组各处死10只大鼠,采用免疫组织化学方法研究雌激素对大鼠咬肌HSP70表达的影响。结果4周时3组大鼠咬肌HSP70的表达无明显差异（P>0.05）;8周时去卵巢组大鼠HSP70的表达下降,与假手术组、雌激素替代组之间的差异有统计学意义（P＜0.05）,而假手术组与雌激素替代组之间的差异无统计学意义（P>0.05）。结论雌激素水平降低会导致大鼠咬肌HSP70的表达下降,雌激素替代疗法可以预防HSP70表达降低。     

The expression of heat shock protein 70 in the dental pulp following trauma

Abstract – Background/Aim: Traumatic dental injuries vary in severity from mild concussion to avulsion. All organisms respond to stress by inducing the synthesis of a group of proteins known generically as heat shock proteins. The activation of these proteins is an essential cellular mechanism designed to protect against a variety of environmental stresses. It is probable that the production of these proteins is increased in the cells of the traumatized dental pulp, however, this has not as yet been demonstrated. The degree of heat shock proteins expression may be related to the severity of the trauma such that estimating the extent of heat shock proteins expression may provide a practicable way of quantifying these injuries. Materials and Methods: Twenty adult male ferrets were divided into four groups. Each maxillary and mandibular right canine was traumatized with the contralateral canines acting as undamaged controls. The trauma was a simulated concussion injury applied using a uniform force. Animals were killed at 24, 48, 72 and 168 h after the injury. The canines were extracted, sectioned and processed for immunohistochemistry using a mouse monoclonal antibody specific for Hsp70. Results: There was a statistically significant difference in Hsp70 staining between traumatized and non‐traumatized teeth only in the group observed 24 h after the trauma. The expression of heat shock proteins form part of the early pulpal response to trauma.     

A comparison of piezosurgery and conventional surgery by heat shock protein 70 expression

The effects of mechanical instruments on the viability of cells are essential in terms of regeneration. There is considerable interest in cell repair following damage mediated by dental surgical procedures. Cells can tolerate stress by expressing heat shock protein 70 (Hsp70). During and after surgical tooth removal, oxidative stress can activate Hsp70 expression proportional to the intensity of the stress signal stimulus to cope with stress. This study examined the expression of Hsp70 as a potential biomarker of immediate postoperative stress in patients undergoing two different surgical procedures of different severity. Expression of Hsp70 both at mRNA and at protein level in the conventional group was two-fold higher than that of the piezo group. This suggests that tooth movement by the piezo method causes relatively lower stress in the alveolar bone. Piezosurgery provides relatively low stress to the patients and this may help cell repair after the surgical procedure. Patients undergoing more aggressive surgery using conventional methods showed a significant increase in Hsp70 in the immediate postoperative period. Therefore, Hsp70 induction can be a potential tool as a prognostic surgical marker.     

Dynamic protein expression patterns during intraoral wound healing in the rat

Wound healing after cleft palate surgery is often associated with impairment of maxillary growth and dento-alveolar development. Wound contraction and scar tissue formation contribute strongly to these effects. In vitro studies have revealed that fibroblasts isolated during different phases of palatal wound healing show phenotypical differences. They change from a quiescent to an activated state and then partly back to a quiescent state. In this study, we evaluated the existence of fibroblast phenotypes at several time-points during palatal wound healing in the rat. Based on cytoskeletal changes (alpha-sma, vimentin, vinculin), integrin expression (alpha1, alpha2, alpha(v) and beta1) and changes in cellularity, we conclude that phenotypically different fibroblast populations are also present during in vivo wound healing. Alpha-sma and the integrin subunits alpha1 and alpha(v) were significantly up-regulated, and vinculin was significantly down-regulated, at early time-points compared to late time-points in wound healing. These changes point to an activated fibroblast state early in wound healing. Later in wound healing, these activated fibroblasts return only partially to the unwounded situation. These results strongly support the idea that different fibroblast populations with specific phenotypes occur in the course of palatal wound healing.     

A fine structural comparison of the healing of incisional wounds of mucosa and skin

Abstract Incisional wounds of the same length and depth were made on tongue and dorsal skin of rats and the fine structural aspects of the responses of these were compared. Comparisons were made between the timing and degree of phagocytosis, the timing and rate of epithelial cell migration, of overall rate of healing, and of reformation of basal lamina and attachment complex in these two tissues. Phagocytic activity in epithelium and connective tissue of mucosa reached higher levels than in skin and the peak of activity in the epithelium of the mucosa was earlier than in the epidermis. Mononuclear phagocytes were active in phagocytosis, neutrophils showed little phagocytic activity-Epithelial migration was found to begin earlier in mucosa than in skin. Epithelialization of the wound and repair of supporting tissue were completed earlier. The same sequence in the renewal of ultrastructural components of the hemidesmosome-attachment complex was found in skin and mucosa, but was completed earlier at the mucosal site. Mechanical and physical as well as physiologic factors may play a role in determining the differing rates of repair in mucosa and skin.     

Expression of heat shock protein 27 (HSP27) in human temporomandibular joint discs of patients with internal derangement

Heat shock proteins (HSPs) are believed to represent a cellular stress response mechanism that protects intracellular proteins from damaging events. Some studies have demonstrated an enhanced expression of large-molecular-weight HSPs in diseased systematic joints. Small heat shock proteins, and among these HSP27, have been studied to a lesser extent. HSP27 has cytoprotective and biosynthetic functions within chondrocytes, and it is an estrogen-associated protein that is under hormonal modulation. To improve understanding at a molecular level of the pathophysiology of certain temporomandibular joint disorders, the authors carried out this immunohistochemical study to assess the presence of HSP27 in human TMJ discs. Twelve adult human TMJ discs (10 diseased and 2 healthy discs) and 5 TMJ fetal human discs were used in this study. Adult discs and TMJ tissues of human fetuses were fixed in 10% buffered formalin. Samples were then processed for histologic examination. Sections were immunohistochemically stained using the streptavidin-biotin detection method. No reaction product for HSP27 in the discs of fetuses was noted. HSP27 was weakly expressed in normal TMJ discs and highly up-regulated in discs of patients showing new vessel formation and chondroid metaplasia. Any correlation between gender and HSP27 was found in the sample, being the up-regulation of HSP27 related mostly to major histopathological changes. This different pattern of HSP27 immunostaining in human TMJ discs detected in the authors' specimens suggests that the expression of this small HSP is functionally modulated. In fact HSP27 up-regulates in internal derangement specimens with major histopathological changes; on the other hand, it is not expressed or only weakly expressed in TMJ discs of fetuses and normal TMJ discs.     

热休克诱导人舌鳞癌Tca8113细胞HSP70表达的研究

目的:研究热休克恢复期人舌癌Tca8113细胞热休克蛋白(HSP70)的表达和热动力学变化,为人舌鳞癌的免疫治疗提供理论基础。方法:通过人舌鳞癌细胞Tca8113在43℃加热30min,运用免疫组化和流式细胞技术检测在各时间点细胞HSP70的表达,各时间点再次以相同条件加热后用MTT法测各组细胞活性,以期了解Tca8113细胞HSP70表达的动态变化。结果:热休克后恢复2h即出现HSP70的表达,8~12h达到高峰,之后逐渐减少,48h后基本恢复至加热2h后的水平,热休克后恢复12h细胞活性最强。结论:HSP70的迅速表达与缓慢降解对维持Tca8113细胞的生理功能有重要作用。为进一步研究HSP70在人舌癌热疗联合免疫治疗初步提供了理论依据。     

A fine structural comparison of the healing of incisional wounds of mucosa and skin.

Incisional wounds of the same length and depth were made on tongue and dorsal skin of rats and the fine structural aspects of the responses of these were compared. Comparisons were made between the timing and degree of phagocytosis, the timing and rate of epithelial cell migration, of overall rate of healing, and of reformation of basal lamina and attachment complex in these two tissues. Phagocytic activity in epithelium and connective tissue of mucosa reached higher levels than in skin and the peak of activity in the epithelium of the mucosa was earlier than in the epidermis. Mononuclear phagocytes were active in phagocytosis, neutrophils showed little phagocytic activity. Epithelial migration was found to begin earlier in mucosa than in skin. Epithelialization of the wound and repair of supporting tissue were completed earlier. The same sequence in the renewal of ultrastructural components of the hemidesmosome-attachment complex was found in skin and mucosa, but was completed earlier at the mucosal site. Mechanical and physical as well as physiologic factors may play a role in determining the differing rates of repair in mucosa and skin.