本文關鍵詞： 痤瘡丙酸桿菌 中藥水提物 瓊脂稀釋法 痤瘡模型　出處：《瀘州醫(yī)學院》2013年碩士論文　論文類型：學位論文

【摘要】：目的：分別提取十九種中藥的水提物，觀察其對痤瘡丙酸桿菌標準株及臨床株的體外抑制作用，從中篩選出對痤瘡丙酸桿菌具有體外抑制作用的中藥水提物，并將篩選出的藥物外用于兔耳痤瘡模型進行療效及不良反應觀察，從而為今后臨床上痤瘡的治療提供一些新的可供選擇的藥物。方法：1．采集痤瘡患者皮損內容物，37°C厭氧培養(yǎng)48小時后，采用耐氧實驗、革蘭氏染色鏡檢、觸媒實驗、硝酸鹽實驗、糖發(fā)酵實驗等系列生化實驗以及ANI厭氧菌鑒定卡，VITEK全自動微生物鑒定儀鑒定為痤瘡丙酸桿菌。2．經查閱文獻[3-4]后選擇臨床上使用頻率較高的19種中藥（黃柏、黃芩、大黃、金銀花、苦參、連翹、地膚子、白頭翁、百花蛇舌草、魚腥草、馬齒莧、旱蓮草、紫草、板藍根、大青葉、蒲公英、車前子、白芷、蛇床子）各20g，分別加水浸泡、煎煮、過濾、濃縮配制成20ml(相當于原藥濃度1g/ml)的中藥水提物。3．配制含不同濃度的19種中藥水提物的改良GAM培養(yǎng)平板，以瓊脂稀釋法檢測各種藥物的MIC，篩選出對痤瘡丙酸桿菌有體外抑菌作用的中藥水提物；同時檢測臨床上治療痤瘡常用的4種抗生素的MIC，以了解痤瘡丙酸桿菌的耐藥情況。4．選取健康雄性家兔（普通級）7只，采用Kligman法造模，將煤焦油外涂于兔耳的雙側耳管開口2cm×2cm處，每日1次，連續(xù)2周，于造模第7天時皮下注射痤瘡丙酸桿菌菌懸液，濃度為1.0×108CFU/ml，每側約50ul。造模成功后以隨機分組方式將14只兔耳分為7組，每組2只，其中6組擦拭中藥水提物，第7組1只兔耳擦拭夫西地酸乳膏，另1只擦拭生理鹽水進行對照，每日1次，分別于第7天、第14天、第21天觀察其療效及不良反應。結果：1．從80例痤瘡患者皮損處分離培養(yǎng)并鑒定出痤瘡丙酸桿菌27例，陽性率為33.75%，性別、年齡無統(tǒng)計學差異。2．黃柏、黃芩、大黃的MIC為25mg/ml；金銀花、苦參的MIC為50mg/ml；連翹的MIC為100mg/ml；地膚子、白頭翁、百花蛇舌草、魚腥草、馬齒莧、旱蓮草、紫草、板藍根、大青葉、蒲公英、車前子、白芷、蛇床子13味中藥在最高濃度200mg/ml進行測試時仍未產生抑菌作用。3．甲硝唑MIC50為16μg/ml，MIC90為128μg/ml，耐藥率為78.57%�？肆置顾豈IC50為4μg/ml， MIC90為16μg/ml，耐藥率為28.57%。阿奇霉素MIC50為2μg/ml，MIC90為8μg/ml，，耐藥率為21.42%。羅紅霉素MIC50為1μg/ml，MIC90為64μg/ml，耐藥率為46.42%。4．兔耳在涂煤焦油7天后，局部增厚，變硬，毛孔變粗大，毛囊口增大并開放隆起，可見白色粉刺及黑色角栓。第14天時可見紅色炎性丘疹及少許膿皰。5．經分別擦拭黃柏、黃芩、大黃、金銀花、苦參、連翹六種中藥水提物治療7天后的兔耳紅腫減輕，部分消退，14天后兔耳厚度變薄，柔軟度有所增加，21天后局部毛囊口角栓及丘疹、粉刺減少變平。未見紅斑、脫屑等不良反應；外用夫西地酸乳膏治療后的兔耳紅腫消退，而擦拭生理鹽水的兔耳與治療前相比未見明顯變化。結論：1．黃柏、黃芩、大黃、金銀花、連翹、苦參六種中藥水提物對痤瘡丙酸桿菌具有良好的體外抑制作用。2．痤瘡丙酸桿菌對甲硝唑、克林霉素、羅紅霉素、阿奇霉素四種抗生素均有不同程度的耐藥，其中甲硝唑耐藥率最高。3.黃柏、黃芩、大黃、金銀花、連翹、苦參六種中藥水提物對實驗兔耳痤瘡模型有治療作用。4．外用黃柏、黃芩、大黃、金銀花、連翹、苦參六種中藥水提物治療兔耳痤瘡模型未觀察到明顯的不良反應。
[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 擄 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 .

【學位授予單位】：瀘州醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R275.9;R-332

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