湿润烧伤膏对糖尿病性溃疡大鼠创面组织晚期糖基化终末产物及其受体表达的影响研究

doi:10.3969/j.issn.1007-9572.2016.26.006

摘要/Abstract

摘要： 目的　探讨湿润烧伤膏（MEBO）干预糖尿病性溃疡创面愈合的作用机制。方法　于2015年6月选取145只12周龄的SPF级健康雄性SD大鼠，采用随机数字表法将其分为空白组（35只）和糖尿病模型组（110只），糖尿病模型组采用腹腔注射链脲佐菌素（STZ）制备糖尿病大鼠模型，8周后选取空白组30只及糖尿病模型组成模大鼠90只，糖尿病模型组成模大鼠再采用随机数字表法分为模型组、MEBO组及贝复济组，每组30只，4组均制备溃疡模型。成功后MEBO组予MEBO纱条、贝复济组予重组牛碱性成纤维细胞生长因子外用溶液浸透纱条、空白组及模型组予0.9%氯化钠溶液纱条局部换药处理，1次/d。分别于创面干预后第3、6、12天，每次随机选取10只大鼠，观察创面愈合速率；取相同位点创面组织，应用酶联免疫吸附试验（ELISA）检测晚期糖基化终末产物（AGEs）、核因子κB（NF-κB）水平，荧光PCR技术检测AGE受体（RAGE）mRNA表达水平，HE染色观察创面组织炎性细胞浸润、成纤维细胞及血管生成情况。结果　造模8周后，糖尿病模型组体质量低于空白组，血糖水平高于空白组（P<0.01）。空白组、模型组、MEBO组和贝复济组第3、6、12天创面愈合速率比较，差异均有统计学意义（P<0.01）；其中模型组、MEBO组、贝复济组创面愈合速率低于空白组，MEBO组、贝复济组创面愈合速率高于模型组；第6、12天时，贝复济组创面愈合速率低于MEBO组（P<0.05）。空白组、模型组、MEBO组和贝复济组第3、6、12天创面组织AGEs水平比较，差异均有统计学意义（P<0.01）；其中模型组、MEBO组、贝复济组创面组织AGEs水平高于空白组，MEBO组、贝复济组创面组织AGEs水平低于模型组（P<0.05）。空白组、模型组、MEBO组和贝复济组第3、6、12天创面组织NF-κB水平比较，差异均有统计学意义（P<0.05）；其中第6、12天时，模型组创面组织NF-κB水平高于空白组，MEBO组创面组织NF-κB水平低于模型组；第12天时，贝复济组创面组织NF-κB水平高于空白组、低于模型组（P<0.05）。空白组、模型组、MEBO组、贝复济组第3、6、12天创面组织RAGE mRNA表达水平比较，差异均有统计学意义（P<0.05）；其中第6、12天时，模型组、贝复济组创面组织RAGE mRNA表达水平高于空白组，MEBO组创面组织RAGE mRNA表达水平低于模型组（P<0.05）。病理结果显示：干预后第12天，与模型组比较，MEBO组和贝复济组能明显促进成纤维细胞及新生毛细血管生长，减少炎性细胞浸润，其中MEBO组更为明显。结论　MEBO能明显促进糖尿病性溃疡创面愈合，其机制可能与调控创面组织AGEs、NF-κB及RAGE mRNA的表达有关。

关键词: 糖尿病, 溃疡, 湿润烧伤膏, 晚期糖基化终末产物, NF-κB

Abstract: Objective　To explore the mechanism of MEBO in the treatment of diabetic skin ulcer.Methods　A total of 145 healthy male SD rats（aged 12 weeks old）in a SPF grade were randomly assigned to blank group（35 rats）and diabetic model group（110 rats）by random number table method in July 2015.Diabetic model was induced by intraperitoneal injection of streptozotocin，8 weeks later，30 rats in blank group were selected，and 90 successful diabetic rat models were equally divided into diabetic model group，MEBO group and BFGF group with 30 rats in each group，the ulcer models were induced.Rats in MEBO group were treated with MEBO gauze，rats in BFGF group were treated with BFGF gauze，rats in blank group and model group were treated with 0.9% sodium chloride injection gauze，the dressing of each case was changed once a day.At the 3rd，6th and 12th day after intervention，10 rats of each group were selected randomly to investigate wound healing rate.The wound tissue of the same site was obtained for test，levels of AGEs and NF-κB were measured by ELISA，the expression level of RAGE mRNA was measured by fluorescent PCR，and the inflammatory cells，fibroblast and new capillary were observed by HE staining.Results　8 weeks after models were established，the body mass of rats in diabetic model group was significantly lower than that in blank group，the blood glucose level of rats in diabetic model group was significantly higher than that in blank group（P<0.01）.At the 3rd，6th and 12th day after intervention，there was significant difference in wound recovery rate among blank group，model group，MEBO group and BFGF group（P<0.01）；the wound recovery rate of model group，MEBO group and BFGF group was lower than that of bland group respectively，and the wound recovery rate of MEBO group and BFGF group was higher than that of model group respectively；at the 6th and 12th day after intervention，the wound recovery rate of BFGF group was lower than that of MEBO group（P<0.05）.At the 3rd，6th and 12th day after intervention，there was significant difference in AGEs level among blank group，model group，MEBO group and BFGF group（P<0.01）.AGEs level of model group，MEBO group and BFGF group was higher than that of blank group respectively，while AGEs level of MEBO group and BFGF group was lower than that of model group（P<0.05）.At the 3rd，6th and 12th day after intervention，there was significant difference in level of NF-κB among blank group，model group，MEBO group and BFGF group（P<0.05）；at the 6th and 12th day after intervention，level of NF-κB of model group was higher than that of blank group and MEBO group respectively；at the 12th day after intervention，level of NF-κB of BFGF group was higher than that of blank group and was lower than that of model group respectively（P<0.05）.At the 3rd，6th and 12th day after intervention，there was significant difference in expression level of RAGE mRNA among bland group，model group，MEBO group and BFGF group（P<0.05）.At the 6th and 12th day after intervention，the expression level of RAGE mRNA of model group and BFGF group was higher than that of blank group respectively，and expression level of RAGE mRNA of MEBO group was lower than that of model group（P<0.05）.Pathological results showed that，at the 12th day after intervention,compared with model group，faster growth of the fibroblast and new capillary and less infiltration of inflammatory cells were found in MEBO group and BFGF group，especially among MEBO group.Conclusion　MEBO could obviously promote the healing of diabetic skin ulcer by regulating the expression of AGEs，NF-κB and RAGE mRNA in wound tissue.

Key words: Diabetes mellitus, Ulcer, MEBO, AGEs, NF-kappa B

李杰辉, 王丽, 张春霞, 狄钾骐, 李辉. 湿润烧伤膏对糖尿病性溃疡大鼠创面组织晚期糖基化终末产物及其受体表达的影响研究[J]. 中国全科医学, 2016, 19(26): 3153-3159.

LI Jie-hui, WANG Li, ZHANG Chun-xia, DI Jia-qi, LI Hui. Effect of MEBO on Advanced Glycosylation End Products and Their Receptors in Wound Tissue of Rats with Diabetic Skin Ulcer[J]. Chinese General Practice, 2016, 19(26): 3153-3159.

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