基于LuxS/AI-2群体感应系统研究疮愈膏促进肛周慢性创面愈合的作用机制

Study on the Mechanism of Chuangyu Ointment in Promoting Perianal Chronic Wound Healing based on LuxS/AI-2 Quorum Sensing System

目的:研究疮愈膏对肛周慢性创面形成的抑制作用及其对创面细菌生物被膜主要调控基因LuxS和信号分子AI-2的影响。方法:采用SD大鼠60只(200±20)g雄性大鼠随机分为普通创面组(n=15),实验组(n=15),阳性对照组(n=15),安慰剂组(n=15);通过背部新鲜创面每日滴加离心粪液构建肛周慢性创面模型。造模成功后,实验组给予疮愈膏外敷,阳性对照组给予莫匹罗星软膏外敷,安慰剂组给予凡士林纱条外敷,普通创面组给予生理盐水纱条外敷,1次/d,持续14 d。于治疗前和治疗后对各组大鼠创面进行记录分析;实时荧光定量聚合酶链式反应(RTPCR)检测生物被膜主要调控基因LuxS的变化;哈氏弧菌测定AI-2信号分子活性。结果:与安慰剂组比较,实验组和阳性对照组大鼠创面面积均明显缩小,差异具有统计学意义(P<0.05);实验组与阳性对照组差异无统计学意义。与安慰剂组对比,实验组和阳性对照组大鼠创面组织提取细菌中LuxS mRNA相对表达量,信号分子AI-2活性均明显降低(P<0.05),实验组和阳性对照组则无明显差异(P<0.05)。结论:疮愈膏能够加速肛周慢性创面的愈合同时抑制创面细菌生物被膜的形成,其机制可能与下调创面细菌LuxS基因的表达及降低其AI-2信号分子活性有关,LuxS/AI-2群体感应系统可能为疮愈膏治疗肛周慢性创面的机制之一。

Objective:To study the inhibitory effect of Chuangyu Cream on the formation of perianal chronic wounds and its effect on the main regulatory gene LuxS and signaling molecule AI-2 of bacterial biofilm in wounds.Methods:60(200±20)g male SD rats were randomly divided into ordinary wound group(n=15),experimental group(n=15),positive control group(n=15),placebo group(n=15);The perianal chronic wound model was constructed by dripping centrifuged fecal fluid into fresh wounds on the back every day.After the model was successfully created,the experimental group was given external application of Chuangyu ointment,and the positive control group was given external application of mupirocin ointment and a placebo.The control group was given external application of Vaseline gauze strips,and the ordinary wound group was given external application of normal saline gauze strips,once/d for 14 days.The wounds of rats in each group were recorded and analyzed before and after treatment;real-time fluorescence quantitative polymerase chain reaction(RT PCR)was used to detect changes in the main regulatory gene LuxS of the biofilm;Vibrio harveyi was used to detect the activity of AI-2 signaling molecules.Results:Compared with the placebo group,the wound area of the rats in the experimental group and the positive control group was significantly reduced,and the difference Statistically significant(P<0.05);the difference between the experimental group and the positive control group was not statistically significant.Compared with the placebo group,the relative expression of LuxS mRNA in bacteria extracted from rat wound tissue in the experimental group and the positive control group,and the signaling molecule AI-2 activity was significantly reduced(P<0.05),and there was no significant difference between the experimental group and the positive control group(P<0.05).Conclusion:Zhuangyu Cream can accelerate the healing of perianal chronic wounds and inhibit the formation of bacterial biofilm on the wounds.The mechanism may be related to down-regulating the expression of the LuxS gene of wound bacteria and reducing the activity of its AI-2 signaling molecule.The LuxS/AI-2 quorum sensing system may be one of the mechanisms by which Chuangyu Cream treats perianal chronic wounds.