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
发布时间：2019-02-12 23:07
【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(P0.01)。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(P0.01);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(P0.05)。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(P0.01);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(P0.05)。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(P0.05);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(P0.05)。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(P0.05);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(P0.05)。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。
[Abstract]:Objective to investigate the mechanism of MEBO (MEBO) on wound healing of diabetic ulcer. Methods in June 2015, 145 SPF grade healthy male SD rats aged 12 weeks were selected and randomly divided into blank group (35 rats) and diabetic model group (110 rats). Diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ). After 8 weeks, 30 rats in the blank group and 90 rats in the diabetic model were selected. The diabetic model rats were randomly divided into three groups: model group, MEBO group and Befuji group with 30 rats in each group. The ulcer models were made in 4 groups. After success, MEBO group was given MEBO gauze, Befuji group was soaked with recombinant bovine basic fibroblast growth factor solution, blank group and model group were treated with 0.9% sodium chloride solution gauze, once a day. Ten rats were randomly selected at the 3rd day after the intervention to observe the wound healing rate. The level of (AGEs), nuclear factor 魏 B (NF- 魏 B) and the expression of AGE receptor (RAGE) mRNA were detected by enzyme linked immunosorbent assay (ELISA) and fluorescence PCR technique. The infiltration of inflammatory cells, fibroblasts and angiogenesis were observed by HE staining. Results after 8 weeks, the body mass of diabetic model group was lower than that of blank group, and the blood glucose level was higher than that of blank group (P0.01). The rate of wound healing in blank group, model group, MEBO group and Befuji group was significantly higher than that in control group (P0.01). The wound healing rate of model group, MEBO group and befuji group was lower than that of blank group. The wound healing rate of MEBO group and Befuji group was higher than that of model group, and the wound healing rate of Befuji group was lower than that of MEBO group at the 6th day (P0.05). There were significant differences in AGEs levels between the blank group, model group, MEBO group and Befuji group on the 12th day (P0.01). The level of AGEs in model group, MEBO group and Befuji group was higher than that in blank group, and the AGEs level in MEBO group and Befuji group was lower than that in model group (P0.05). There were significant differences in NF- 魏 B levels between the blank group, model group, MEBO group and Befuji group on the 12th day (P0.05). On the 6th day, the level of NF- 魏 B in the model group was higher than that in the blank group, the level of NF- 魏 B in the MEBO group was lower than that in the model group, and on the 12th day, the NF- 魏 B level in the wound tissue in the befuji group was higher than that in the blank group and lower than that in the model group (P0.05). In blank group, model group, MEBO group and Befuji group, the expression of RAGE mRNA in wound tissue was significantly higher than that in control group (P0.05). At the 6th day, the expression of RAGE mRNA in the model group and Befuji group was higher than that in the blank group, and the expression level of RAGE mRNA in the MEBO group was lower than that in the model group (P0.05). The pathological results showed that on the 12th day after intervention, MEBO group and Befuji group could significantly promote fibroblast and neovascularization growth, and reduce inflammatory cell infiltration, especially in MEBO group, compared with the model group. Conclusion MEBO can obviously promote wound healing of diabetic ulcer, and its mechanism may be related to regulating the expression of AGEs,NF- 魏 B and RAGE mRNA in wound tissue.
【作者单位】： 广西中医药大学第一附属医院;广西壮族自治区中医药研究院;
【基金】：国家自然科学基金资助项目(81302975)
【分类号】：R587.2
本文编号：2420888
[Abstract]:Objective to investigate the mechanism of MEBO (MEBO) on wound healing of diabetic ulcer. Methods in June 2015, 145 SPF grade healthy male SD rats aged 12 weeks were selected and randomly divided into blank group (35 rats) and diabetic model group (110 rats). Diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ). After 8 weeks, 30 rats in the blank group and 90 rats in the diabetic model were selected. The diabetic model rats were randomly divided into three groups: model group, MEBO group and Befuji group with 30 rats in each group. The ulcer models were made in 4 groups. After success, MEBO group was given MEBO gauze, Befuji group was soaked with recombinant bovine basic fibroblast growth factor solution, blank group and model group were treated with 0.9% sodium chloride solution gauze, once a day. Ten rats were randomly selected at the 3rd day after the intervention to observe the wound healing rate. The level of (AGEs), nuclear factor 魏 B (NF- 魏 B) and the expression of AGE receptor (RAGE) mRNA were detected by enzyme linked immunosorbent assay (ELISA) and fluorescence PCR technique. The infiltration of inflammatory cells, fibroblasts and angiogenesis were observed by HE staining. Results after 8 weeks, the body mass of diabetic model group was lower than that of blank group, and the blood glucose level was higher than that of blank group (P0.01). The rate of wound healing in blank group, model group, MEBO group and Befuji group was significantly higher than that in control group (P0.01). The wound healing rate of model group, MEBO group and befuji group was lower than that of blank group. The wound healing rate of MEBO group and Befuji group was higher than that of model group, and the wound healing rate of Befuji group was lower than that of MEBO group at the 6th day (P0.05). There were significant differences in AGEs levels between the blank group, model group, MEBO group and Befuji group on the 12th day (P0.01). The level of AGEs in model group, MEBO group and Befuji group was higher than that in blank group, and the AGEs level in MEBO group and Befuji group was lower than that in model group (P0.05). There were significant differences in NF- 魏 B levels between the blank group, model group, MEBO group and Befuji group on the 12th day (P0.05). On the 6th day, the level of NF- 魏 B in the model group was higher than that in the blank group, the level of NF- 魏 B in the MEBO group was lower than that in the model group, and on the 12th day, the NF- 魏 B level in the wound tissue in the befuji group was higher than that in the blank group and lower than that in the model group (P0.05). In blank group, model group, MEBO group and Befuji group, the expression of RAGE mRNA in wound tissue was significantly higher than that in control group (P0.05). At the 6th day, the expression of RAGE mRNA in the model group and Befuji group was higher than that in the blank group, and the expression level of RAGE mRNA in the MEBO group was lower than that in the model group (P0.05). The pathological results showed that on the 12th day after intervention, MEBO group and Befuji group could significantly promote fibroblast and neovascularization growth, and reduce inflammatory cell infiltration, especially in MEBO group, compared with the model group. Conclusion MEBO can obviously promote wound healing of diabetic ulcer, and its mechanism may be related to regulating the expression of AGEs,NF- 魏 B and RAGE mRNA in wound tissue.
【作者单位】： 广西中医药大学第一附属医院;广西壮族自治区中医药研究院;
【基金】：国家自然科学基金资助项目(81302975)
【分类号】：R587.2
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，本文编号：2420888
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