中药水提物对痤疮丙酸杆菌体外抑制作用及对兔耳痤疮模型的疗效研究

发布时间：2018-01-19 04:31

本文关键词： 痤疮丙酸杆菌中药水提物琼脂稀释法痤疮模型　出处：《泸州医学院》2013年硕士论文　论文类型：学位论文

【摘要】：目的：分别提取十九种中药的水提物，观察其对痤疮丙酸杆菌标准株及临床株的体外抑制作用，从中筛选出对痤疮丙酸杆菌具有体外抑制作用的中药水提物，并将筛选出的药物外用于兔耳痤疮模型进行疗效及不良反应观察，从而为今后临床上痤疮的治疗提供一些新的可供选择的药物。方法：1．采集痤疮患者皮损内容物，37°C厌氧培养48小时后，采用耐氧实验、革兰氏染色镜检、触媒实验、硝酸盐实验、糖发酵实验等系列生化实验以及ANI厌氧菌鉴定卡，VITEK全自动微生物鉴定仪鉴定为痤疮丙酸杆菌。2．经查阅文献[3-4]后选择临床上使用频率较高的19种中药（黄柏、黄芩、大黄、金银花、苦参、连翘、地肤子、白头翁、百花蛇舌草、鱼腥草、马齿苋、旱莲草、紫草、板蓝根、大青叶、蒲公英、车前子、白芷、蛇床子）各20g，分别加水浸泡、煎煮、过滤、浓缩配制成20ml(相当于原药浓度1g/ml)的中药水提物。3．配制含不同浓度的19种中药水提物的改良GAM培养平板，以琼脂稀释法检测各种药物的MIC，筛选出对痤疮丙酸杆菌有体外抑菌作用的中药水提物；同时检测临床上治疗痤疮常用的4种抗生素的MIC，以了解痤疮丙酸杆菌的耐药情况。4．选取健康雄性家兔（普通级）7只，采用Kligman法造模，将煤焦油外涂于兔耳的双侧耳管开口2cm×2cm处，每日1次，连续2周，于造模第7天时皮下注射痤疮丙酸杆菌菌悬液，浓度为1.0×108CFU/ml，每侧约50ul。造模成功后以随机分组方式将14只兔耳分为7组，每组2只，其中6组擦拭中药水提物，第7组1只兔耳擦拭夫西地酸乳膏，另1只擦拭生理盐水进行对照，每日1次，分别于第7天、第14天、第21天观察其疗效及不良反应。结果：1．从80例痤疮患者皮损处分离培养并鉴定出痤疮丙酸杆菌27例，阳性率为33.75%，性别、年龄无统计学差异。2．黄柏、黄芩、大黄的MIC为25mg/ml；金银花、苦参的MIC为50mg/ml；连翘的MIC为100mg/ml；地肤子、白头翁、百花蛇舌草、鱼腥草、马齿苋、旱莲草、紫草、板蓝根、大青叶、蒲公英、车前子、白芷、蛇床子13味中药在最高浓度200mg/ml进行测试时仍未产生抑菌作用。3．甲硝唑MIC50为16μg/ml，MIC90为128μg/ml，耐药率为78.57%。克林霉素MIC50为4μg/ml， MIC90为16μg/ml，耐药率为28.57%。阿奇霉素MIC50为2μg/ml，MIC90为8μg/ml，，耐药率为21.42%。罗红霉素MIC50为1μg/ml，MIC90为64μg/ml，耐药率为46.42%。4．兔耳在涂煤焦油7天后，局部增厚，变硬，毛孔变粗大，毛囊口增大并开放隆起，可见白色粉刺及黑色角栓。第14天时可见红色炎性丘疹及少许脓疱。5．经分别擦拭黄柏、黄芩、大黄、金银花、苦参、连翘六种中药水提物治疗7天后的兔耳红肿减轻，部分消退，14天后兔耳厚度变薄，柔软度有所增加，21天后局部毛囊口角栓及丘疹、粉刺减少变平。未见红斑、脱屑等不良反应；外用夫西地酸乳膏治疗后的兔耳红肿消退，而擦拭生理盐水的兔耳与治疗前相比未见明显变化。结论：1．黄柏、黄芩、大黄、金银花、连翘、苦参六种中药水提物对痤疮丙酸杆菌具有良好的体外抑制作用。2．痤疮丙酸杆菌对甲硝唑、克林霉素、罗红霉素、阿奇霉素四种抗生素均有不同程度的耐药，其中甲硝唑耐药率最高。3.黄柏、黄芩、大黄、金银花、连翘、苦参六种中药水提物对实验兔耳痤疮模型有治疗作用。4．外用黄柏、黄芩、大黄、金银花、连翘、苦参六种中药水提物治疗兔耳痤疮模型未观察到明显的不良反应。
[Abstract]:Objective : To observe the in vitro inhibitory effect of nineteen Chinese herbs on acne Propionibacterium standard strain and clinical strain , and to screen out the traditional Chinese medicine water extract with in vitro inhibitory effect on Propionibacterium acne , and to observe the effect and adverse reaction of the screened drug on the rabbit ear acne model , so as to provide some new alternative medicine for the treatment of acne in the future . After 48 hours after 37 掳 C anaerobic culture , the skin lesion contents of patients with acne were collected , and the VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 VITEK 2 Systems for Identification of Propionibacterium Propionibacterium with a series of biochemical experiments such as oxygen - resistant experiments , Gram staining microscopy , catalyst experiments , nitrate experiments , sugar fermentation experiments and the like . 20 g of 19 Chinese medicinal materials ( Cortex Phellodendri , Scutellariae Radix , Radix et Rhizoma Rhei , Flos Lonicerae , Radix Sophorae Flavescentis , Fructus Forsythiae , Kochiae Fructus , Radix Sophorae Flavescentis , Fructus Forsythiae , Herba Houttuyniae , Radix Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Angelicae Dahuricae , and Fructus Cnidii , respectively , soaking in water , decocting , filtering , and concentrating to obtain 20 ml of Chinese medicinal water extract . The invention relates to a method for preparing a traditional Chinese medicine water extract with different concentrations of 19 Chinese medicinal water extracts , which is used for detecting the MIC of various drugs by an agar dilution method , and screening the traditional Chinese medicine water extract which has in vitro antibacterial effect on Propionibacterium acne ; and simultaneously , the MIC of the four antibiotics commonly used for treating acne is detected , so as to know the drug resistance of Propionibacterium acne . A healthy male rabbit ( normal grade ) 7 was selected and the coal tar was applied to rabbit ears at 2 cm 脳 2 cm twice a day for 2 weeks . The concentration was 1.0 脳 108 CFU / ml , and the concentration was 1.0 脳 108 CFU / ml . 27 cases of Propionibacterium were isolated and cultured from 80 patients with acne . The positive rate was 33.75 % . There was no statistical difference between sex and age . MIC of Cortex Phellodendri , Scutellariae Radix and Radix et Rhizoma Rhei is 25 mg / ml ; MIC of Flos Lonicerae and Radix Sophorae Flavescentis is 50 mg / ml ; the MIC of Fructus Forsythiae is 100 mg / ml ; Fructus Kochiae , Radix Paeoniae Alba , Herba Thymi , Radix Isatidis , Folium Isatidis , Herba Taraxaci , Semen Plantaginis , Radix Angelicae Dahuricae , and Fructus Cnidii are tested at the highest concentration of 200 mg / ml . The MIC50 of metronidazole was 16渭g / ml , MIC90 was 128 渭g / ml , the drug resistance rate was 78.57 % . The MIC50 was 4渭g / ml , MIC90 was 16渭g / ml , the drug resistance rate was 28.57 % . The MIC50 of azithromycin was 8渭g / ml , MIC90 was 6渭g / ml , MIC90 was 64渭g / ml , and the resistance rate was 46.42 % . After 7 days of application of coal tar , the rabbit ears thickened locally , hardened , the pores became coarse , the pores of the hair follicle enlarged and opened , and white pimples and black horns were visible . Red inflammatory papules and little pustules were visible at the 14th day . After 7 - day treatment of 7 - day old rabbit ear - swelling was relieved after 7 - day treatment with six kinds of Chinese medicinal water extracts , such as Cortex Phellodendri , Scutellariae Radix , Radix et Rhizoma Rhei , Flos Lonicerae , Radix Sophorae Flavescentis , and Fructus Forsythiae . The traditional Chinese medicine water extract of Cortex Phellodendri , Scutellariae Radix , Radix et Rhizoma Rhei , Flos Lonicerae , Fructus Forsythiae , and Radix Sophorae Flavescentis has good in vitro inhibitory effect against Propionibacterium acne . The resistance rate of metronidazole was the highest . 3 . The drug resistance rate of metronidazole was the highest . 3 . There were six kinds of traditional Chinese medicine water extracts , such as Cortex Phellodendri , Scutellariae Radix , Radix et Rhizoma Rhei , Flos Lonicerae , Fructus Forsythiae , and Radix Sophorae Flavescentis , and has therapeutic effect on experimental rabbit ear acne model . no obvious adverse reaction was observed in the treatment of the rabbit ear acne model with six kinds of Chinese medicinal water extracts , such as Cortex Phellodendri , Scutellariae Radix , Radix et Rhizoma Rhei , Flos Lonicerae , Fructus Forsythiae , and Radix Sophorae Flavescentis .

【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R275.9;R-332

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