湿润烧伤膏对糖尿病大鼠创面组织细胞间黏附分子1和血管细胞黏附分子1 mRNA表达水平及超微结构的影响研究

Effect of MEBO on ICAM-1 mRNA and VCAM-1 mRNA Expression Levels and Ultrastructure in Wound Tissue of Rats with Diabetic Ulcer

目的探讨湿润烧伤膏(MEBO)干预糖尿病性溃疡创面组织细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)m RNA表达及超微结构的作用机制。方法于2015年6月选取145只12周龄的雄性健康SPF级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)法制备糖尿病大鼠模型,8周后选取空白组30只和糖尿病模型组成模大鼠90只,采用随机数字表法将糖尿病模型组成模大鼠分为模型亚组、贝复济亚组及MEBO亚组,各30只,均制备糖尿病性溃疡创面模型。模型制备成功后,空白组及模型亚组给予0.9%氯化钠溶液纱条、贝复济亚组给予贝复济溶液纱条、MEBO亚组给予MEBO纱条局部换药处理,1次/d,共12 d。各组分别于创面干预治疗后第3、6、12天,每次随机选取10只大鼠,取相同位点创面组织,应用荧光聚合酶链式反应(PCR)技术检测ICAM-1、VCAM-1 m RNA表达水平,电镜观察创面组织超微病理结构。结果模型亚组第3天创面组织ICAM-1 m RNA表达水平低于空白组,第6、12天创面组织ICAM-1 m RNA表达水平均高于空白组(P<0.05);贝复济亚组第12天创面组织ICAM-1 m RNA表达水平低于模型亚组(P<0.05);MEBO亚组第3天创面组织ICAM-1 m RNA表达水平高于模型亚组,第6、12天创面组织ICAM-1m RNA表达水平均低于模型亚组(P<0.05)。模型亚组第3天创面组织VCAM-1 m RNA表达水平低于空白组,第6、12天创面组织VCAM-1 m RNA表达水平均高于空白组(P<0.05);贝复济亚组、MEBO亚组第3天创面组织VCAM-1m RNA表达水平均高于模型亚组,第6、12天创面组织VCAM-1 m RNA表达水平均低于模型亚组(P<0.05)。病理结果显示:干预后第6、12天,与模型亚组比较,贝复济亚组和MEBO亚组细胞结构均逐步改善,能促进各细胞器形态恢复正常。结论 MEBO能够调控糖尿病性溃疡创面组织中糖基化终末产物(AGEs)-AGEs受体(RAGE)信号通路下游因子ICAM-1、VCAM-1的表达,改善细胞超微结构,促进糖尿病性溃疡创面愈合。

Objective To explore the effect and mechanism of MEBO on ICAM-1 mRNA,VCAM-1 mRNA expression levels and ultrastructure in wound tissue of rats with diabetic ulcer.Methods A total of 145 12-week-old healthy male SD rats of SPF grade were randomly assigned to blank group(n=35)and diabetic model group(n=110)by the random digital table in June 2015.Diabetic model was induced by intraperitoneal injection of streptozotocin(STZ).Eight weeks later,30 rats in blank group and 90 diabetic rats(equally divided into model subgroup,bFGF subgroup and MEBO subgroup)were selected for establishing diabetic ulcer models.Then,both the blank group and model subgroup,bFGF subgroup,and MEBO subgroup received local treatment of the ulcer with normal saline-moistened gauze,bFGF-moistened gauze,and MEBOmoistened gauze,respectively.The dressing was changed once a day for 12 days.At 3,6 and 12 days after intervention,10 rats were selected randomly from each group for taking the wound tissue from the same site for test.The expression levels of ICAM-1 mRNA and VCAM-1 mRNA were measured by fluorescent PCR and the ultrastructural pathology of the wound tissue was observed by electron microscopy.Results Compared with the blank group,the mean expression level of ICAM-1 mRNA in the model subgroup was lower at 3 days after intervention,but higher at 6 and 12 days after intervention(P<0.05).The expression level of ICAM-1 mRNA in the bFGF subgroup was significantly lower than model subgroup at 12 days after intervention(P<0.05).The expression level of ICAM-1 mRNA in the MEBO subgroup was higher than that in the model subgroup at 3 days after intervention(P<0.05),but it was lower than that in the model subgroup at 6 and 12 day s after intervention(P<0.05).Compared with the model subgroup,blank group presented higher mean VCAM-1 mRNA expression level at 3 days after intervention,but lower mean VCAM-1 mRNA expression level at 6,12 days after intervention(P<0.05),as did both bFGF and MEBO subgroups(P<0.05).Pathological results showed that at 6 and 12 days after intervention,in both MEBO and bFGF subgroups,the cell structure of the wound tissue was improved gradually,contributing to the recovery of organelle morphology,while in the model subgroup,similar status was not found.Conclusion MEBO could obviously promote the healing of diabetic ulcer by regulating the expression of downstream factors ICAM-1 and VCAM-1 mRNA in AGEs-RAGE signaling pathway and improving cell ultrastructure in wound tissue.