人血红蛋白及其片段体内外抗菌活性研究

目的分离人血红蛋白及其片段,进行体外抗菌活性分析和比较,并对血红蛋白体内抗菌活性进行初步探讨。方法利用阳离子交换及分子筛分离人血红蛋白α、β链;溴化氰法裂解α/β链,反相高效液相色谱技术纯化其片段;琼脂糖弥散法抑菌实验检测抗菌活性;构建大鼠大肠埃希菌阴道感染模型,设立实验组(血红蛋白组)和对照组,观察各组大鼠阴道组织形态学变化。结果体外抑菌实验示:血红蛋白、α/β链及其片段有抑菌作用,但其主要针对的是革兰阴性菌大肠埃希菌;而且除α1~32的抑菌作用较弱外,血红蛋白、α/β链及其片段三者抑菌活性比较,其效能基本相似。大鼠体内抗菌实验示:与基质对照组(感染后不治疗)比,血红蛋白组阴道复层扁平上皮表层较平滑,固有层内炎细胞浸润明显减少,且组织内充血明显减轻。结论人血红蛋白及其片段具有体外抑菌活性,而且血红蛋白在大鼠体内能减轻大肠埃希菌阴道感染炎症程度。

Study on Antimicrobial Activity of Human Hemoglobin and Its Fragments both in vitro and in vivo

OBJECTIVE: To isolate human hemoglobin and its fragrments, compare their antimicrobial activity in vitro and pilot study of their antimicrobial activity in vivo. METHODS: The alpha and beta chains of hemoglobin were separated by cation exchange chromatography and gel chromatography; The alpha and beta chains were cleaved by cyanogens bromide respectively. The cleaved fragments were purified by reverse phase high performance liquid chromatography. Antimicrobial activity of hemoglobin and its fragments was determined by agrose radial diffusion assay. After establishment of E. coli vaginal infection model, the rats were randomized into the experimental group (hemoglobin group) and the control group. The histologically pathological section was observed. RESULTS: Hemoglobin, alpha/beta chain and their fragments had similar antibacterial activities in vitro, which were mainly against Gram-negative bacteria E. coli; except alpha1-32 had a comparatively lower activity. Antimicrobial activity in vivo: a comparison of the hemoglobin group with the matrix control group (no treatment after infection), the surface layer of vaginal stratified squamous epithelium was smoother, inflammatory cells were significantly reduced in the lamina propria and congestion was obviously decreased. CONCLUSION: Human hemoglobin and its fragments had antibacterial activity in vitro, hemoglobin might relieve the inflammation of E. coil vaginal infection in rats moreover.