Adenovirus-mediated METH1 gene expression inhibits hypertrophic scarring in a rabbit ear model

Hypertrophic scarring remains a major problem for patients who have suffered from surgeries or burns. Vascularization plays an important role in the early phase of hypertrophic scarring. Therefore, the inhibition of angiogenesis might be used as a preventive strategy. In this study, we assessed the effect of anti-angiogenesis resulting from adenovirus-mediated METH1 (metalloprotease and thrombospondin1) gene expression on the hypertrophic scar formation in a rabbit ear model of hypertrophic scarring. We first investigated the number of microvessel and microcirculatory perfusion in untreated scars on days 10, 30, 60, and 90 after epithelialization. Then, we examined the effect of anti-angiogenesis by adenovirus-mediated METH1 expression on hypertrophic scar formation by calculating the scar elevation index, counting the microvessel and argyrophilic nucleolar organizer region particle, and detecting the amount of collagen on days 30 and 60 after treatment. We found that untreated scar tissues at the proliferative phase (days 10–60 after epithelialization) had a significantly higher density of microvessel and microcirculatory perfusion than those at the mature phase (day 90 after epithelization) (both p <0.05). On days 30 and 60 after treatment, the hypertrophic scar formation was significantly inhibited in the treatment group. There was significantly reduced scar elevation index, microvessel count, number of argyrophilic nucleolar organizer region, and total collagen content for treated scars. Our results demonstrate that METH1 has a markedly inhibitive effect on the formation of hypertrophic scar, and may thus have a promising application in the prevention of human hyperthropic scars.     

ALA-PDT对兔耳增生性瘢痕抑制效应及作用机制的初步研究

目的：观察ALA-PDT（5-氨基酮戊酸光动力疗法）对兔耳增生性瘢痕形成的抑制作用及对转化生长因子β1（TGF-β1）表达的影响。方法：在兔耳腹侧面做直径1cm大小全层皮肤缺损创面3个/侧,共60个。随机分为以下三组：高浓度ALA-PDT组（n=20）：20%ALA介导PDT治疗;低浓度ALA-PDT组（n=20）：10%ALA介导PDT治疗;对照组（n=20）：不做治疗,待创面自然愈合。在创面形成术后的第6天、第13天分别进行ALA-PDT治疗,观察治疗组与对照组之间的瘢痕形成率、瘢痕增生指数、胶原纤维含量及TGF-β1表达情况的差异。结果：ALA-PDT治疗降低了瘢痕形成率,其中药物高浓度ALA-PDT组与对照组的差异有统计学意义（P〈0.01）;药物高浓度ALA-PDT组和药物低浓度ALA-PDT组的瘢痕增生指数、胶原纤维含量与对照组的差异有统计学意义（P〈0.05）;药物高浓度ALA-PDT组与对照组之间的TGF-β1的表达差异具有统计学显著性（P〈0.05）。结论：ALA-PDT对兔耳增生性瘢痕的形成有抑制作用,从而预防创伤后增生性瘢痕形成,该作用可能通过减少TGF-β1的产生而实现。     

Smad ubiquitination regulatory factor 2 expression is enhanced in hypertrophic scar fibroblasts from burned children

Transforming growth factor-β1 (TGF-β1) plays a key role in hypertrophic scar formation. A lot of studies have shown that TGF-β1 stimulates fibroblast proliferation, collagen production, and α-smooth muscle actin (α-SMA) expression, inhibits matrix degradation and eventually leads to scar formation. Smad proteins are important intracellular mediators of TGF-β1 signaling, and Smad ubiquitination regulatory factor 2 (Smurf2), an ubiquitin ligase for Smads, plays critical roles in the regulation of TGF-β1/Smad signaling. It was reported that Smurf2 was abnormally expressed during the process of liver fibrosis and lung fibrosis. Hypertrophic scarring is a fibroproliferative disorder of the dermis that occurs following wounding. However, little is known about the expression of Smurf2 in hypertrophic scarring. We hypothesized that TGF-β1 signaling cannot be disrupted after wound epithelialization probably due to abnormal expression of Smurf2 in hypertrophic scar fibroblasts. In the present study, we found that hypertrophic scar fibroblasts exhibited increased Smurf2 protein and mRNA levels compared with normal fibroblasts, and the expression of Smurf2 gradually increased in hypertrophic scar fibroblasts after TGF-β1 stimulation. Furthermore, we transfected Smurf2 siRNA into hypertrophic scar fibroblasts, and we found that silencing the expression of Smurf2 in hypertrophic scar fibroblasts dramatically reduced TGF-β1 production, inhibited TGF-β1-induced α-SMA expression and inhibited TGF-β1-induced collagen I synthesis. Our results suggest that the enhanced expression of Smurf2 is involved in the progression of hypertrophic scarring.     

The role of Th1/Th2 cell chemokine expression in hypertrophic scar

The aim of this study was to study the role of Th1/Th2 cell‐associated chemokines in the formation of hypertrophic scars in rabbit ears. Twenty‐six New Zealand white rabbits were used to establish the hypertrophic scar model of rabbit ear and the normal scar model of rabbit's back. Two rabbits were sacrificed on days 0 and 21, 28, 35, 42, 49, 56, and 63 after operation. The specimens were stained with haematoxylin‐eosin (HE). Scar elevation index (SEI) was used to detect the expression of 10 chemokines related to Th1/Th2 cells in both scar formation expressions. Real‐time polymerase chain reaction (PCR) results showed that two chemokines (CXCL10, CXCL12) were highly expressed during the formation of normal scar, and there was almost no expression during the formation of hypertrophic scar (*P < 0.05). The chemokines (CCL2, CCL3, CCL4, CCL5, CCL7, CCL13, CX3CL1) were almost non‐expressed in the formation of normal scars but were expressed for a long time in the formation of hypertrophic scars. The four chemokines, CCL2, CCL4, CCL5, and CX3CL1, maintained a long‐term high expression level during the formation of hypertrophic scars (P < 0.01). There were also three chemokines (CCL14, CCL19, CCL21) that were almost undetectable in normal scarring, but there was transiently low‐level expression (P < 0.05) only during the peak proliferative phase in proliferative scarring. Th1/Th2 cell‐associated chemokines are different in the type, quantity and expression, and maintenance time of rabbit ear hypertrophic scars.     

15-脱氧-△^12,14-前列腺素J2对兔耳增生性瘢痕的作用

目的探讨γ过氧化物酶体增殖物激活受体（peroxisome proliferator activated receptor-7,PPAR-γ）的配体15-脱氧-△^12,14-前列腺素J2（15-deoxy-△^12,14-prostagliandxinJ2,15d—PGJ2）对兔耳增生性瘢痕I型胶原、结缔组织生长因子（connective tissue growth factor,CTGF）、α平滑肌肌动蛋白（a—smooth muscle actin,α—SMA）表达的影响,探讨15d—PGJ2防治增生性瘢痕的可能性。方法选取新西兰大白兔18只,在兔耳腹侧面制作2cm×3cm全层皮肤缺损创面,每耳2个,共计72个,建立兔耳增生性瘢痕动物模型,随机分为2组,一组为实验组,另一组为对照组,分别用15d—PGJ2和生理盐水行瘢痕内注射,1次／d,共7次。停药后第7、14、21天两组同时取材;每组每次切取12个组织块。应用免疫组织化学检测I型胶原、CTGF和α—SMA的表达。结果各组兔耳腹侧创面愈合后均不同程度出现类似人增生性瘢痕的组织块。与对照组相比,15dPGJ2注射后瘢痕体积缩小,变软变平,色泽轻度变浅。在各个时间点15d—PGJ2组I型胶原、CTGF和α-SMA的表达均较对照组低,且差异有统计学意义（P〈0．05）。结论PPAR-γ的配体15d—PGJ2可降低瘢痕内I型胶原、CTGF和α—SMA的含量,引起瘢痕萎缩,有一定防治瘢痕的作用,可能为临床治疗增生性瘢痕提供一条新的途径。     

烧伤后增生性瘢痕患者血清中TGF-β和VEGF的浓度检测及意义

目的:探讨烧伤后增生性瘢痕患者血清中TGF-β、VEGF浓度变化及意义。方法:选择2009,8~2011,8我科住院的烧伤后增生性瘢痕患者共74例,根据创面愈合后瘢痕增生时间长短分组,分为4组,选择12例正常人作为正常对照组,共5组。用酶联吸附法(ELISA)检测患者血清中TGF-β、VEGF的浓度,5组之间统计比较分析。结果:瘢痕患者血清中TGF-β、VEGF含量与瘢痕增生时段关系密切,1~3月早期瘢痕组血清中TGF-β、VEGF含量开始增加,随瘢痕的发展升高,4~6个月瘢痕组血清中TGF-β、VEGF含量达到了高峰。随着瘢痕成熟,TGF-β、VEGF浓度又逐渐下降。结论:烧伤后增生性患者血清中TGF-β、VEGF的含量可较好地反映增生性瘢痕代谢活跃程度,是增生性瘢痕增生活跃程度敏感的标记物。     

Temporal expression of the transforming growth factor-Beta pathway in the rabbit ear model of wound healing and scarring

BACKGROUND: Despite numerous studies that have investigated the cellular and molecular mechanisms underlying scar formation, this process still remains poorly understood. The importance of transforming growth factor-beta (TGF-beta) in these processes has been well recognized, and this study sought to define the temporal expression of the key members in this pathway in a well-established, clinically relevant, rabbit ear model of hypertrophic scarring. STUDY DESIGN: Seven-millimeter (hypertrophic) and 5-mm (nonhypertrophic) punch wounds were made on the ears of 12 rabbits. Wounds were harvested at days 0, 7, 15, 28, and 40. RESULTS: There were no appreciable histologic differences between the 5- and 7-mm wounds at days 7 and 15. At day 28, however, the 7-mm scars were considerably more hypertrophic compared with the 5-mm control scars (p<0.001). The mRNA levels of TGF-beta1 and collagen Ialpha2 were notably higher in the hypertrophic 7-mm scars at day 28 than in the nonhypertrophic 5-mm scars (p<0.03). Although not pronounced, levels of TGF-beta2 were higher in the hypertrophic scars. There were no other statistically significant differences between the 7- and 5-mm scars. CONCLUSIONS: Elevated levels of TGF-beta1, and possibly TGF-beta2, are associated with hypertrophic scar formation.     

Basic fibroblast growth factor (bFGF) alleviates the scar of the rabbit ear model in wound healing

Studies suggest a possible antiscarring effect of basic fibroblast growth factor (bFGF) during wound healing. However, little is known about the precise pathological mechanisms of bFGF. In particular, there is only limited information available about the mechanism of exogenous administration of bFGF to scar formation. To investigate the effect of bFGF on the hypertrophic scar in the rabbit ear model and to clarify the mechanisms of bFGF on treatment for scar in wound healing, the rabbit ear model of wound healing was created and treated topically with bFGF once daily for 3 months; then we examined the changes of macroscopic and histopathological characteristics of scars and the expression of collagen and collagenase-1 (matrix metalloproteinase-1). The results of macroscopic and histologic characteristics revealed a significant difference between scars treated with bFGF and control scars. The expression of collagen in the scars treated with bFGF was decreased, as compared with the scars treated with saline. Further study revealed that bFGF could remarkably enhance expression of matrix metalloproteinase-1. bFGF could improve the quality of wound healing and remarkably alleviate the scar in the rabbit ear model in wound healing, which suggests that bFGF exerted a net negative effecton scar formation in wound healing. The evidence should contribute to a better understanding of the biological activities of bFGF during hypertrophic scar formation.     

Recombinant human decorin inhibits TGF-β1-induced contraction of collagen lattice by hypertrophic scar fibroblasts

Decorin was reported to bind transforming growth factor-beta (TGF-β₁) and neutralise some of its activity as a key regulator of wound contraction and hypertrophic scar formation. In this study, we investigated whether recombinant human decorin affected TGF-β₁-induced fibroblast contractile activity, by using fibroblast-populated collagen lattice with decorin added to the collagen gel. Hypertrophic scar fibroblasts showed greater basal contraction of collagen gels than normal fibroblasts, and the addition of TGF-β₁ significantly enhanced this. Decorin inhibited both the basal and TGF-β₁-enhanced contraction of collagen gel by both normal and hypertrophic scar fibroblasts. Decorin also inhibited TGF-β₁-induced α-smooth muscle actin (α-SMA), plasminogen activator inhibitor-1 (PAI-1) protein and mRNA expressions in normal and hypertrophic scar fibroblasts. These results suggest that decorin may have therapeutic potential for excessive skin contraction as observed in hypertrophic scarring.     

Pigment epithelium‐derived factor attenuates angiogenesis and collagen deposition in hypertrophic scars

Scar forming wounds are often characterized by higher levels of vascularity than non‐scarring wounds and normal skin, and inhibition of angiogenesis has been shown to inhibit scar formation in some model systems. The rabbit ear hypertrophic scar (HS) model has been widely used to study the mechanisms that underlie the development of HS as well as the effectiveness of various treatments. Although the rabbit ear HS model is well characterized in terms of scar formation, the rate and level of angiogenesis has not been investigated in this model, and the cause‐effect relationship between angiogenesis and rabbit HSs has not been examined. In the current study, full‐thickness excisional wounds were created on the ventral side of New Zealand White rabbit ears to induce HS formation, and the dynamic pattern of angiogenesis and the expression of angiogenic regulatory factors were examined over time. Blood vessel density was found to peak at 2.7% on day 14 post‐wounding, decreasing to 1.7% by day 28. mRNA levels of the proangiogenic factor VEGF‐A peaked at day 14, while the expression of the antiangiogenic factors pigment epithelium‐derived factor (PEDF) and thrombospondin 1 (TSP1) peaked at day 28 post‐wounding. To examine whether inhibition of angiogenesis influences HS formation in this model, wounds were treated with exogenous soluble antiangiogenic agents including recombinant PEDF (rPEDF) and a functional PEDF peptide (PEDF‐335). rPEDF and PEDF‐335 were administered intradermally from day 4 post‐wounding every 3 days until day 19. Intradermal injection of rPEDF or PEDF‐335 both led to decreased angiogenesis and decreased collagen deposition at the wound site. The results support the utility of antiangiogenic therapies, including rPEDF/PEDF‐335, as a potential new strategy for the prevention or treatment of HSs.     

曲安奈德、干扰素α-2b和维拉帕米治疗增殖瘢痕的作用机制比较

目的 比较曲安奈德、干扰素α-2b和维拉帕米局部注射对瘢痕疙瘩和增生性瘢痕增殖、凋亡和TGF-β1表达的影响. 方法 增生性瘢痕和瘢痕疙瘩各6例,局部注射曲安奈德(40 mg/ml)、干扰素α-2b(150万U/ml)和维拉帕米(2.5 mg/ml)后7 d,切取标本,采用免疫组织化学、末端脱氧核苷酸介导的生物素化的脱氧尿嘧啶DNA切口末端标记方法,检测细胞增殖核抗原和TGF-β1的表达及细胞发生的凋亡情况,并以未注射药物的瘢痕疙瘩和增生性瘢痕以及健康皮肤为对照. 结果 ①曲安奈德可抑制瘢痕疙瘩和增生性瘢痕细胞增殖和诱导细胞凋亡,同时抑制细胞TGF-β1表达从而抑制瘢痕的增殖增生;②干扰素α-2b可通过抑制瘢痕疙瘩、增生性瘢痕细胞的增殖和TGF-β1表达而抑制瘢痕的增殖增生,但其不能诱导细胞凋亡;③维拉帕米可通过抑制瘢痕疙瘩、增生性瘢痕细胞的增殖和诱导细胞凋亡而抑制瘢痕的增殖,同时抑制细胞TGF-β1表达,其诱导细胞凋亡的作用妹强于曲安奈德,但抑制TGF-β1表达作用弱于曲安奈德和干扰素α-2b. 结论 曲安奈德、干扰素α-2b和维拉帕米局部注射后,对瘢痕疙瘩和增生性瘢痕在临床上虽均有效,但作用机制不尽相同.     

Expression of vascular endothelial growth factor and its receptor Flk-1 in human neuroblastoma using in situ hybridization

Background/Purpose: Although angiogenic factors may play an important role in the biology of neuroblastoma, which frequently spreads hematogenously, the mechanism remains unclear. The authors studied tumor progression and invasion from the perspective of angiogenesis and sought to understand the features of this type of tumor. Methods: Thirty-one specimens were resected from patients with neuroblastoma and the expression of vascular endothelial growth factor (VEGF), and its receptor (Flk-1) was examined using immunohistochemistry. The authors looked for correlations among the expressions of VEGF and its receptor with various clinicopathologic factors. In addition, they examined the expression and location of VEGF and Flk-1 mRNA in 10 primary neuroblastoma using in situ hybridization. Results: Both in situ hybridization and immunohistochemistry showed the presence of VEGF expression within the neuroblastoma cells. We found VEGF mRNA in neuroblastoma cells but not vascular endothelial cells according to in situ hybridization. Further, Flk-1 mRNA was present both in neuroblastoma cells and vascular endothelial cells. The level of VEGF expression was higher in unfavorable histology, using the criteria of Shimada, than in favorable histology. Conclusion: The authors suggest that paracrine and autocrine systems are involved in the angiogenesis of neuroblastoma, and the expression of VEGF correlates with the prognosis in neuroblastoma.     

Smad7 down-regulation via ubiquitin degradation mediated by Smurf2 in fibroblasts of hypertrophic scars in burned patients

TGF-β1 (transforming growth factor β1) was considered to play a critical role in the forming of hypertrophic scars. Smad, as a kind of signal downstream mediators, can modulate the functions of TGF-β1. Smad7 can regulate TGF-β1/Smad pathway and present negative feedbacks, which prevents fibrosis mediated by TGF-β1. Nonetheless, the mechanisms related to Smad7 activity in regulating hypertrophic scarring are hardly known. The studies have shown that Smad7 decrease induced by the increase of Smurf2 (Smad ubiquitination regulatory factor 2, an E3 ubiquitin ligase of Smad7) ubiquitination degradation plays a part in fibrosis. We thus made a hypothesis that Smad7 could not inhibit TGF-β1 because Smurf2 ubiquitin degradation was increased in hypertrophic scar fibroblasts. In our research, it was discovered that there was an increase in Smad7 mRNA levels but no increase in Smad7 protein levels in the fibroblasts of hypertrophic scars after TGF-β1 treatment. The ubiquitination activity and degradation of Smad7 protein were increased in the fibroblasts of hypertrophic scars compared with the fibroblasts of normal skin. Enhanced degradation of Smad7 protein in the fibroblasts of hypertrophic scars was prevented by proteasome inhibitors MG132 / MG115. Furthermore, it was found that TGF-β1 stimulation increased Smad7 protein expression after silencing Smurf2 gene in hypertrophic scar fibroblasts, and enhanced Smad7 degradation was prevented in hypertrophic scar fibroblasts after Smurf2 was silenced. It was implied that ubiquitin degradation mediated by Smurf2 might contribute to decreased Smad7 protein levels following TGF-β1 stimulation in the fibroblasts of hypertrophic scars.     

瘢痕内注射脂肪来源干细胞对兔耳增生性瘢痕的抑制作用研究

目的 研究瘢痕内注射脂肪来源干细胞(ADSC)对兔耳增生性瘢痕的抑制作用及可能机制。方法 选取12只新西兰大白兔,制作兔耳增生性瘢痕模型,随机平均分成3组,2周后各组右耳瘢痕内注射DMEM作为自身对照,左耳分别注射ADSCs、脂肪来源干细胞条件培养基(ADSCs-CM)及不作处理。注射前及注射后1、2、3周检测瘢痕增生情况,并于注射后3周取材,行组织化学及基因学检测。结果 造模2周后,伤口均完全上皮化;DMEM注射及未处理组的伤口逐渐出现增厚、变红、变硬等增生表现,注射后3周时最为明显;ADSCs及ADSCs-CM注射组均未出现明显增生反应。取材后HE、Masson染色示,ADSCs及ADSCs-CM注射组瘢痕的胶原密度适中、排列整齐;对照及未处理组瘢痕可见大量致密、杂乱的胶原组织。基因检测发现,ADSCs及ADSCs-CM注射组瘢痕的α-SMA、CollagenⅠ表达显著低于对照及未处理组。ADSCs注射组瘢痕冰冻切片Dil荧光染色可见大量存活脂肪来源干细胞。结论 瘢痕内注射脂肪来源干细胞可以降低α-SMA、CollagenⅠ基因表达,从而改善瘢痕内胶原堆积,并最终改善瘢痕增生情况。     

15-脱氧-△12,14-前列腺素J2对兔耳增生性瘢痕Ⅰ型胶原表达的影响

目的 观察过氧化物酶体增殖物激活受体γ(PPARγ)的配体15-脱氧-△12,14-前列腺素J2(15d-PGJ2)对兔耳增生性瘢痕Ⅰ型胶原表达的影响,探讨15d-PGJ2防治增生性瘢痕的可行性.方法 选取新西兰大白兔18只,在兔耳腹侧面制作2 cm×3 cm全层皮肤缺损创面,每耳2个,共计72个,建立兔耳增生性瘢痕动物模型,随机分组,分别用15d-PGJ2及生理盐水行瘢痕内注射,1次/d,共7次.停药后第14、21天两组同时取材;每组每次切取18个组织块.应用免疫组织化学、荧光定量聚合酶链反应(PCR)及Western blot检测Ⅰ型胶原的表达.结果 与对照组比较,15d-PGJ2注射后瘢痕体积缩小,变软变平,色泽轻度变浅.Ⅰ型胶原主要分布于真皮的细胞间质、成纤维细胞胞质中,血管壁上亦见阳性信号,在各个时间点15d-PGJ2组Ⅰ型胶原mRNA和蛋白的表达均较对照组低,且差异有统计学意义(P<0.05).结论 PPAR-γ的配体15d-PGJ2可降低瘢痕内Ⅰ型胶原的含量,引起瘢痕萎缩,从而防治瘢痕.     

A role of cytokines in OK-432 injection therapy for cystic lymphangioma: an approach to the mechanism

Purpose: This study examines cytokine levels of aspirates from cystic lymphangiomas after injection of OK-432 and over the course of several weeks to better understand the process of tumor regression.Methods: Fluids aspirated from lymphangioma cysts of 3 patients were collected sequentially before and after OK-432 injection. Mononuclear cells (MNCs) were separated and cultured with or without OK-432. Vascular endothelial growth factor (VEGF), soluble VEGF receptor 1 (sVEGFR1), sVEGFR2, transforming growth factor beta-1 (TGF-β1), interleukin (IL)-6, IL-8, IL-12+p40, tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) levels in the supernatants of the aspirates and the culture supernatants of MNCs were then measured by ELISA.Results: The aspirates exhibited a marked elevation in IL-6, IL-8, VEGF, and TGF-β1 levels for a few weeks after the OK-432 injection. IL-6, IL-8, IL-12+p40, TNF-α, and IFN-γ levels were elevated in the culture supernatants of the MNC cultured with OK-432 for up to 9 days. All the tumors regressed significantly, with sclerotic change, within 3 months after OK-432 injection.Conclusions: Cytokine production is maintained for a few weeks after OK-432 injection. Fibrotic changes may be another main mechanism in tumor regression in addition to cytotoxic effects on lymphangioma cells. A close relationship between cytokines from intracystic cells and lymphangioma cells is suggested.     

黄芪对家兔增生性瘢痕转化生长因子β1表达及其Smad 3信号途径的影响

目的 观察黄芪对家兔增生性瘢痕TGF-β1及其Smad 3信号表达的影响,分析它对增生性瘢痕的治疗作用及机制. 方法 选用日本大耳白兔20只,在每只兔单耳腹侧致伤4处形成瘢痕组织.按照随机数字表法分为1.00、0.50、0.25 g/mL黄芪治疗组(于伤后21、25、32、36 d分别注射黄芪0.2 mL),生理盐水组(同上时相点注射等量生理盐水),空白对照组(未作任何处理),每组32个瘢痕部位.另选4只大耳白兔作为正常对照组.组织病理学方法 观察瘢痕组织结构变化;彩色超声诊断仪、瘢痕硬度精密测定仪分别测定伤后32、43 d瘢痕厚度和硬度;RT-PCR、免疫组织化学法观察瘢痕组织中TGF-βt、Smad 3 mRNA水平和蛋白的变化.数据处理采用t检验、单因素方差分析.结果 伤后32、43 d各黄芪治疗组兔耳瘢痕真皮层均较生理盐水组、空白对照组变薄,其中伤后43 dFb、胶原纤维排列较整齐.黄芪治疗组瘢痕组织的厚度、硬度,TGF-β1、Smad 3 mRNA和蛋白水平随着药物浓度的降低呈上升趋势.伤后32 d 1.00 g/mL黄芪治疗组TGF-β1、Smad 3 mRNA水平分别为生理盐水组的73.9%、71.8%;蛋白水平分别为3.15±0.80、4.72±1.06,明显低于生理盐水组(6.06±0.85、8.04±0.63,F值分别为27.230、33.525,P＜0.05或P＜0.01).各黄苠治疗组除TGF-β1、Smad 3 mRNA外,其他指标在伤后32、43 d 2个时相点比较,差异均有统计学意义[t值分别为3.593～4.814(厚度)、4.051～5.811(硬度)、2.976～5.986(TGF-βt蛋白水平)、2.742～4.630(Smad 3蛋白水平),P＜0.05或P＜0.01]. 结论 黄芪注射液通过抑制瘢痕组织中Fb增殖,减少TGF-β1、Smad 3 mRNA表达水平和蛋白合成,从而抑制兔耳瘢痕组织增生,其抑制作用与用药浓度、时间呈剂量效应关系,可能成为今后治疗增生性瘢痕选择方法 之一.     

血管抑制因子METH1基因转染对兔耳瘢痕组织增生的影响

目的 研究血管抑制因子METH1基因转染对兔耳瘢痕成纤维细胞增殖、胶原合成的影响.方法 复制兔耳增生性瘢痕模型,用微循环显微镜检、瘢痕组织AgNOR染色、苦味酸一天狼星红染色等方法,观察基因重组血管抑制剂Ad-METH1对增生性瘢痕组织血管生成、成纤维细胞增殖、胶原分布的影响.结果 Ad-METH1注射后30 d瘢痕组织血管生成明显减少,成纤维细胞增殖减弱、Ⅰ/Ⅲ型胶原比明显降低.结论 上皮化后早期血管抑制基因治疗可有效抑制增生性瘢痕的形成.