外源性高迁移率族蛋白B1对大鼠烫伤早期创面缺血带血管生成的影响

代蕾; 郭杏; 黄海峻; 廖筱梅; 罗兴前; 李丹; 周虹; 高小春; 谭美云

doi:10.3760/cma.j.issn.1009-2587.2018.04.006

外源性高迁移率族蛋白B1对大鼠烫伤早期创面缺血带血管生成的影响

doi: 10.3760/cma.j.issn.1009-2587.2018.04.006

1. 646000 四川省泸州市,西南医科大学附属第一医院整形烧伤外科;
2. 646000 四川省泸州市,西南医科大学附属第一医院骨关节外科

基金项目:

四川省医学会课题（160832）

国家自然科学基金（31271049）

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出版历程
- 收稿日期: 2017-06-30
- 网络出版日期: 2021-10-28
- 刊出日期: 2018-04-20

Effects of exogenous high mobility group protein box 1 on angiogenesis in ischemic zone of early scald wounds of rats

1. Department of Burns and Plastic Surgery, the First Affiliated Hospital of Southwest Medical University, Luzhou 646000, China

摘要

摘要: 目的观察外源性高迁移率族蛋白B1(HMGB1)对大鼠烫伤早期创面缺血带血管生成的影响。方法取36只SD大鼠，按随机数字表法分为HMGB1组和单纯烫伤组，每组18只。将定制铜质梳状模具置于100 ℃沸水中3～5 min后，置于大鼠背部20 s，造成有3个缺血带的创面。烫伤后即刻，HMGB1组大鼠从创面缺血带边缘皮下注射0.4 μg HMGB1＋0.1 mL磷酸盐缓冲液(PBS)，单纯烫伤组大鼠同前注射0.1 mL PBS。烫伤后24、48、72 h，每组各取6只大鼠，采用免疫组织化学法检测烫伤后24、48、72 h创面缺血带血管内皮生长因子(VEGF)蛋白表达量及烫伤后48、72 h创面缺血带CD31蛋白表达量。显微镜下观察并计算烫伤后48、72 h创面缺血带组织CD31免疫组织化学切片中微血管数量。实时荧光定量反转录PCR检测烫伤后24、48、72 h创面缺血带VEGF、CD31 mRNA表达量。对数据行析因设计方差分析、t检验及Bonferroni校正。结果 (1)烫伤后24、48、72 h，HMGB1组大鼠创面缺血带VEGF蛋白表达量明显高于单纯烫伤组(t＝7.496、4.437、5.402，P<0.05或P<0.01)。烫伤后48、72 h，HMGB1组大鼠创面缺血带CD31蛋白表达量分别为0.038 8±0.007 9、0.057 7±0.001 2，明显高于单纯烫伤组的0.013 4±0.004 9、0.030 3±0.004 0(t＝10.257、15.055，P<0.01)。(2)烫伤后48、72 h，HMGB1组大鼠创面缺血带微血管数明显多于单纯烫伤组(t＝3.536、4.000，P<0.05)。(3)烫伤后24、48、72 h，HMGB1组大鼠创面缺血带VEGF mRNA表达量明显高于单纯烫伤组(t＝4.406、3.821、3.356，P<0.05)。烫伤后24、48 h，HMGB1组大鼠创面缺血带CD31 mRNA表达量明显高于单纯烫伤组(t＝4.113、3.466，P<0.05)；烫伤后72 h，2组大鼠创面缺血带CD31 mRNA表达量相近(t＝0.010，P>0.05)。结论外源性HMGB1可通过提高VEGF和CD31的表达，从而促进大鼠烫伤早期创面缺血带血管生成。
- 高迁移率族蛋白质类 /
- 新生血管化，病理性 /
- 血管内皮生长因子类 /
- CD31
Abstract: Objective To observe effects of exogenous high mobility group protein box 1 (HMGB1) on angiogenesis in ischemic zone of early scald wounds of rats. Methods Thirty-six Sprague-Dawley rats were divided into HMGB1 group and simple scald (SS) group according to the random number table, with 18 rats in each group. Comb-like copper mould was placed on the back of rats for 20 s after being immersed in 100 ℃ hot water for 3 to 5 min to make three ischemic zones of wound. Immediately after scald, rats in HMGB1 group were subcutaneously injected with 0.4 μg HMGB1 and 0.1 mL phosphate buffer solution (PBS), and rats in SS group were subcutaneously injected with 0.1 mL PBS from boarders of ischemic zone of scald wound. At post scald hour (PSH) 24, 48, and 72, 6 rats in each group were collected. Protein expressions of vascular endothelial growth factor (VEGF) in ischemic zone of wound at PSH 24, 48, and 72 and protein expressions of CD31 in ischemic zone of wound at PSH 48 and 72 were detected by immunohistochemistry. The number of microvessel in CD31 immunohistochemical sections of ischemic zone of wound at PSH 48 and 72 was calculated after observing by the microscope. The mRNA expressions of VEGF and CD31 in ischemic zone of wound were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction at PSH 24, 48, and 72. Data were processed with analysis of variance of factorial design, t test, and Bonferroni correction. Results (1) At PSH 24, 48, and 72, protein expressions of VEGF in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=7.496, 4.437, 5.402, P<0.05 or P<0.01). At PSH 48 and 72, protein expressions of CD31 in ischemic zone of wound of rats in HMGB1 group were 0.038 8±0.007 9 and 0.057 7±0.001 2 respectively, significantly higher than 0.013 4±0.004 9 and 0.030 3±0.004 0 of rats in SS group (t=10.257, 15.055, P<0.01). (2) At PSH 48 and 72, the number of microvessel in ischemic zone of wound of rats in HMGB1 group was obviously more than that of rats in SS group (t=3.536, 4.000, P<0.05). (3) At PSH 24, 48, and 72, mRNA expressions of VEGF in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=4.406, 3.821, 3.356, P<0.05). At PSH 24 and 48, mRNA expressions of CD31 in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=4.113, 3.466, P<0.05). At PSH 72, mRNA expressions of CD31 in ischemic zone of wound of rats in 2 groups were close (t=0.010, P>0.05). Conclusions Exogenous HMGB1 can promote angiogenesis in ischemic zone of early scald wounds of rats by increasing expressions of VEGF and CD31.
- High mobility group proteins /
- Neovascularization, pathologic /
- Vascular endothelial growth factors /
- CD31