透脓散对脓肿模型大鼠抗菌、免疫功能与局部病理变化作用机理研究

Abscesses can occur in subcutaneous and visceral, and antibiotic drugs are the first choice for the treatment.But there are many problems of antibiotics for the treatment of abscess, that has become a bottleneck in the treatment of abscess and suppurative inflammation, it received widespread attention in the medicine and the whole society, and try to seek solutions.. TouNongSan was famous special prescription to treat abscess in TCM. In addition to the widely used in the carbuncle, suppurative mastitis and perianal abscess, also used in liver and kidney abscess, Acute suppurative appendicitis.but little research on its action mechanism.The project members study found TouNongSan could improve the healing rate of local abscess and bFGF and EGF levels of the rats that was infected aureus bacteria in primary study.This research further use agar dilution method to test the antibacterial effect in vitro and in vivo;and use the model rats with subcutaneous abscess formed by staphylococcus aureus, then the rats were took corresponding Tounongsan,Cefradine, physiological saline after grouping.We draw blood and peritoneal macrophages from the rats in according to the prescribed time,test Mac-1 and CR3 expression of phagocytic cells by flow cytometry, the phagocytosis and bactericidal function by bacteriolytic methods and phagocytosis test, serum total complement activity and cytokine by ELISA, single complement by immune nephelometry, proliferation and secretion of B lymphocyte function by enzyme linked immune spot assay;At the same time detect pathological changes and cell infiltration in inflammation local tissues by microscope.. The project aims to systematically test and find the TouNongSan mechanism, and to further study in the future,develop the antibacterial medicine.

脓肿可发生于皮下和内脏，药物治疗首选抗菌素。不过抗菌素在治疗脓肿存在诸多问题，已受到医药界普遍关注，并努力寻求解决途径。透脓散作为中医治疗脓肿的著名专用方剂，临床中除了广泛用于痈、化脓性乳腺炎与肛周脓肿等浅部化脓性疾病，还用于肝肾脓肿、化脓性阑尾炎等内脏化脓性疾病。可使脓肿早日成熟、破溃，病灶由深及浅，趋于局限，轻者吸收消散。然而关于其作用机理研究甚少。项目成员前期研究发现透脓散可提高金葡菌感染大鼠局部脓肿的愈合率与bFGF、EGF水平。本项目拟进一步研究透脓散体内外抑菌作用；用金葡菌感染大鼠致局部脓肿模型，观察其对血与腹腔液中吞噬细胞Mac-1和 CR3表达、吞噬和杀菌功能影响，检测血清总补体活性、细胞因子、补体与B淋巴细胞增殖与分泌功能，分析对免疫功能作用；镜下检测炎症局部组织病理变化与细胞浸润情况。本项目系统试验旨在发现透脓散多方面作用机制，便于今后深入研究、开发抗菌中药。

1.项目的背景.脓肿可发生于皮下和内脏，药物治疗首选抗菌素。不过抗菌素在治疗脓肿存在诸多问题，已受到医药界普遍关注。透脓散作为中医治疗脓肿的著名专用方剂，临床中除了广泛用于痈、化脓性乳腺炎与肛周脓肿等浅部化脓性疾病，还用于肝肾脓肿、化脓性阑尾炎等内脏化脓性疾病。其可使脓肿早日成熟、破溃，病灶由深及浅，趋于局限，轻者吸收消散。本项目拟进一步研究透脓散多方面作用机理，便于今后深入研究、开发抗菌中药。.2.主要研究内容.（1）透脓散水提液及醇提液成分含量测定与对比。.（2）透脓散体外、体内抑菌试验。.（3）对中性粒细胞和巨噬细胞细胞表面CR3、吞噬功能作用。.（4）对补体、细胞因子的影响。.（5）对体液免疫的作用（淋巴细胞增殖与分泌功能）。.（6）局部取材，HE染色、免疫组化标记染色，观测细胞浸润、血管增生与组织修复。.3.重要结果、关键数据其科学意义.（1）透脓散水提液及醇提液含量测定.醇提液中主要成分含量黄芪甲苷＞总黄酮＞阿魏酸＞槲皮素，醇提液中黄芪甲苷、总黄酮含量高于水提液，70%甲醇及70%乙醇提液中其他两种成分与水提液相近。.（2）透脓散体内外抑菌实验结果.体外抑菌实验，2种提取液对金黄色葡萄球菌、乙型溶血性链球菌、肺炎链球菌有明显抑制作用，且醇提取液较水提取液在后两种菌上抑制作用明显。体内抑菌实验，透脓散水提液高剂量组、头孢羟氨苄组与病模组比较死亡率明显降低（P＜0.05），水提液高剂量组与头孢羟氨苄组比较无明显差异。.科学意义 透脓散体内、体外均有显著的抑菌作用，并且主要对革兰阳性球菌抑制明显，这将对透脓散的合理使用有重要的指导意义。.（3）对中性粒细胞FC受体、C3b受体影响实验结果.透脓散能升高中性粒细胞C3b受体水平，且给药6 d，优于头孢拉定组；给药3 d，能升高中性粒细胞FC受体水平；并且可以影响血C5a水平，具有双向调节作用。.科学意义 透脓散组能升高中性粒细胞C3b、FC受体水平，促进中性粒细胞黏附与吞噬，双向调节血中C5a水平。.（4）由于疫情影响，实验已全部完成，部分数据正在分析。