摘要
目的:通过电镜观测及Toll样受体4(TLR-4)、IL-1β的检测,明确丹黄消炎液对创面感染耐药鲍曼不动杆菌的抑制作用及其机理。方法:新西兰白兔30只,采用皮肤切开注菌法建立皮肤感染泛耐药鲍曼不动杆菌溃疡模型,随机分为三组:空白组、中药灌胃组、中药外敷组。空白组创面外敷0.9%氯化钠溶液,同时0.9%氯化钠溶液10 mL/kg灌胃1次。中药灌胃组创面外敷0.9%氯化钠溶液,同时丹黄消炎液10 mL/kg每日灌胃1次。中药外敷组创面丹黄消炎液外敷,同时0.9%氯化钠溶液10 mL/kg每日灌胃1次。分别于造模成功后用药前、用药后第24、48、72 h及第10 d取各组伤口处组织及第72 h分泌物,以Western blotting检测伤口处组织TLR-4表达,免疫组化检测IL-1β含量。扫描电镜检测分泌物中鲍曼不动杆菌形态。结果:用药后48 h中药灌胃组TLR-4为0.1809±0.0357,IL-1β为2.0503±0.2632,中药外敷组TLR-4为0.1824±0.0194,IL-1β为2.6405±0.5425,表达均较空白组明显升高(P<0.05),用药后72 h中药灌胃组TLR-4为0.1501±0.0060,IL-1β为1.8793±0.9265,中药外敷组TLR-4为0.1540±0.0028,IL-1β为1.9195±0.5334,表达均较空白组明显升高(P<0.05),且用药48 h TLR-4、IL-1β达到峰值。用药后10d空白组TLR-4、IL-1β表达明显高于中药灌胃组、中药外敷组(P<0.05)。不同用药途径丹黄消炎液作用对TLR-4、IL-1β无明显差异(P>0.05)。用药后72 h电镜检测显示,中药灌胃组、中药外敷组细菌大量坏死粘连,中药外敷组略优于中药灌胃组。结论:中药制剂丹黄消炎液对感染泛耐药鲍曼不动杆菌溃疡有抑制作用,可能与TLR-4、IL-1β的表达升高参与机体的炎症反应调节有关;不同用药途径与丹黄消炎液作用是否有影响,值得进一步研究。
Objective To determine the inhibitory effect of Danhuang Xiaoyan Liquid(DXL)on the infection of Acinetobacter Bauman and its mechanism by electron microscope observation and detection of TLR-4and IL-1β.Methods Thirty New Zealand white rabbits were divided into three groups:blank group,gavage group of traditional Chinese medicine(TCM)and TCM plastering group.The skin infection pan resistant Acinetobacter Bauman ulcers model was established using skin incision and injection of bacteria.In the blank group,0.9%of sodium chloride solution was applied to the wound and0.9%of sodium chloride solution(10mL/kg)was given once daily.Sodium chloride solution(0.9%)in the TCM gavage group was treated with external application of Sodium Chloride Solution,while the DXL10mL/kg was gavaged once daily.The TCM plastering group was treated with external application of DXL,and0.9%sodium chloride solution(10mL/kg)was gavaged once daily.After the model was successfully established,the tissues of each group were obtained at24h,48h,72h after treatment,the wound secretion was also taken at72h after treatment.The level of TLR-4was detected by Western blotting,and the content of IL-1βwas detected by immunohistochemistry.The morphology of Acinetobacter Baumannii in secretions was detected by scanning electron microscope(SEM).Results After48h,TLR-4of TCM gavage group was0.1809±0.0357,IL-1βwas2.0503±0.2632.TLR-4of TCM plastering group was0.1824±0.0194,IL-1βwas2.6405±0.5425.There expression in the two groups was significantly higher than that in the blank group(P<0.05).After72h of Treatment,TLR-4of TCM gavage group was0.1501±0.0060,IL-1βwas1.8793±0.9265.But for TCM plastering group,the expression of TLR-4was0.1540±0.0028,and IL-1βwas1.9195±0.5334,both of which were significantly higher than those in the blank group(P<0.05).The peak values of TLR-4and IL-1βwere reached at48h after treatment.The expression of TLR-4and IL-1βon day10in blank group was significantly higher than that in the TCM gavage group and TCM plastering group(P<0.05).The effect of DXL on expression of TLR4and IL-1βwas not significantly different(P>0.05).The electron microscope examination at72h after treatment showed that the bacteria in the TCM gavage group and the TCM plastering group were largely necrotic and adhesive,and the TCM plastering group was slightly superior to the TCM gavage group.Conclusion The decoction of TCM,DXL,can inhibit the infection of Pan-resistant Acinetobacter bauman,which may be related to the regulation of TLR-4and IL-1βexpression and the regulation of inflammatory reaction.
作者
徐阳
王军
薛田
张荣荣
沈凤娇
XU Yang;WANG Jun;XUE Tian(First Teaching Hospital of Tianjin University of TraditionalChinese Medicine, Tianjin (300193), China)
出处
《中国中西医结合外科杂志》
CAS
2018年第6期723-728,共6页
Chinese Journal of Surgery of Integrated Traditional and Western Medicine
基金
国家自然科学基金青年项目(81403394)
关键词
丹黄消炎液
耐药鲍曼不动杆菌
TOLL样受体4
电镜检测
Danhuang Xiaoyan Liquid
resistant Acinetobacter Baumannii
toll-like receptor 4
electronmicroscope examination
作者简介
通信作者:王军,E-mail:tjzywangjun@126.com。
