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目的观察大鲵皮肤分泌液中抗菌肽对铜绿假单胞菌感染小鼠创面的抗菌作用。方法用5%醋酸浸提和SephadexG-50、G-25凝胶过滤层析等方法分离抗菌肽;采用Tricine-SDS-PAGE电泳鉴定,抑菌圈法检测抗菌活性;切除30只ICR小鼠背部1 cm×1 cm全层皮肤,涂抹铜绿假单胞菌菌液制成感染模型,随机均分为对照组、磺胺米隆组和抗菌肽组,伤后3 h分别用生理盐水、100 g.L-1磺胺米隆、0.5 g.L-1抗菌肽的纱布湿敷。伤前和伤后3 d抽血观察白细胞的变化;伤后每天观察小鼠创面的感染情况、精神状态和体重变化;14 d后观察小鼠的存活情况。结果检测到大鲵皮肤分泌液中抗菌肽的相对分子质量约为4.3×103 Da,对铜绿假单胞菌的抗菌活性较强;术后3组小鼠的白细胞计数均减少;对照组小鼠相比其他两组活动减少,创面潮湿、分泌物较多,磺胺米隆组、抗菌肽组的创面结痂、干燥、无明显分泌物;对照组小鼠的体重在第6天开始下降,明显少于其他两组(P<0.05);而无菌对照组的小鼠无死亡,重量持续增加。14 d后对照组存活率明显少于磺胺米隆组和抗菌肽组(P<0.05)。结论大鲵皮肤分泌液中抗菌肽对铜绿假单胞菌感染的ICR小鼠创面有较强的抗感染作用,可明显降低其死亡率。
Objective To observe the antibacterial effect of antibacterial peptides on the wound surface of Pseudomonas aeruginosa infected mice. Methods Antibacterial peptides were separated by 5% acetic acid extraction, Sephadex G-50 and G-25 gel filtration chromatography. Antibacterial activity was detected by Tricine-SDS-PAGE electrophoresis. 1 cm × 1 cm full-thickness skin, smear Pseudomonas aeruginosa bacterial liquid infection model, were randomly divided into control group, sulfamethazine group and antibacterial peptide group, 3 h after injury, respectively, with saline, 100 gL- 1 sulfamethamine, 0.5 gL-1 antibacterial peptide gauze wet compress. The changes of white blood cells were observed before and 3 days after injury. The wound infection, mental status and body weight were observed daily after injury. The survival of mice was observed after 14 days. Results The relative molecular mass of the antimicrobial peptides in the skin exudates was 4.3 × 103 Da and the antibacterial activity against Pseudomonas aeruginosa was stronger. The white blood cell count of the three groups of mice decreased after operation. The number of mice in the control group Compared with the other two groups, the activity was reduced, the wound was wet and the secretions were more. The wounds of sulfamethoxine group and antimicrobial peptide group were scabbed, dry and had no obvious secretions. The body weight of control mice began to decrease on the 6th day, In the other two groups (P <0.05); while the mice in the sterile control group showed no death, and their weight continued to increase. After 14 days, the survival rate of the control group was significantly less than the sulfamethazine group and antimicrobial peptide group (P <0.05). Conclusion The antibacterial peptides in the skin exudates of Dachagtan can strongly inhibit the wounds of ICR mice infected with Pseudomonas aeruginosa and can significantly reduce the mortality rate.