复方金银花中药漱口水的制备及其功效实验研究

发布时间：2018-05-03 07:13

本文选题：牙周炎 + 漱口水　；参考：《南方医科大学》2017年硕士论文

【摘要】：研究背景及目的牙周炎是造成我国成年人牙齿丧失的主要原因。目前,大多数学者认为,牙周炎是多因素疾病,其中牙菌斑生物膜的细菌及其毒性产物是引发牙周炎必不可少的始动因子。因此,控制和去除牙菌斑是牙周炎治疗的关键。菌斑控制的常用方法为机械性菌斑控制法,包括龈上洁治、龈下刮治、刷牙等。在基础治疗的基础上,以含有抗菌成分的漱口水作为辅助手段,能取得更好的菌斑控制效果。漱口水在口腔疾病的预防及辅助治疗中起着重要的作用。根据功能及成分的不同,漱口水可分为防龋类漱口水、抗敏感类漱口水、抗生素类漱口水、抑制牙菌斑类漱口水、口气清新类漱口水及中草药类漱口水,在清除口腔异味、防龋齿、抗炎等方面有很好的效果,常作为牙周炎、牙龈炎、冠周炎及口腔溃疡等疾病的辅助用药。其中,抑制牙菌斑类漱口水作为自我菌斑控制手段的一种,可通过液体流动力进入牙刷、牙线、洁治器等器械不能到达的区域,补充机械性菌斑控制法的不足。目前,临床上常用的漱口水中多含有抗生素、碘、乙醇等成分,如洗必泰、含碘新型漱口水等,长期使用易引起牙面着色、口腔菌群失调、口腔黏膜刺激等问题。因此,寻找有效、安全、无酒精、可长期使用的漱口水成为牙周病防治的新热点。在我国,天然药物的应用历史悠久、且毒副作用小,取材方便,经济实惠,有的还具有杀菌消炎作用,因而广泛用于基础研究及临床实践。中药制剂在口腔中的运用广泛,大量研究表明中药在治疗牙周病、口腔溃疡等疾病上具有良好的疗效。但目前对于该类中药的报道多为某种单一中草药的作用,对中药复方的研究较少,而中药复方更能体现中医辨证论治的特点,充分发挥药物的协同作用及中医药多种有效成分多靶点作用。随着人们对漱口水安全性的关注,中药类漱口水的优势逐渐显现出来。市面上已有成品销售的中药类漱口水,但普遍存在色泽浑浊、口感较差或含有酒精等问题,使得其接受度较其他类漱口水低。且对于中草药类漱口水的研究,从基础到应用的系统研究较少,缺乏科学客观的疗效观察,且对于其毒性及不良反应报道较少,未能给临床应用提供实验依据。中医理论认为胃火炽热、肾气虚亏和气血不足是牙周炎的主要病因。茵陈、野菊花、金银花、白芷、细辛生物活性强,被证实具有抗炎、调节免疫等作用,且对包括口腔致病菌在内的多种细菌具有抑制作用。将这5种药物进行复方配伍,应用于防治牙周疾病中尚未见报道。综上所述,本研究拟从茵陈、野菊花、金银花中提取单味药液,并提取细辛+白芷混合药液,通过均匀设计法获得5种药物的最佳配比,制备复方金银花中药漱口水,通过动物学实验,研究该漱口水的毒性及对牙周疾病的功效,为进一步开发出安全有效的复方金银花中药漱口水提供科学依据。研究方法1.采用挥发油提取法对茵陈、野菊花、金银花、细辛、白芷5种药物进行提取,制备茵陈、野菊花、金银花单味药液及细辛+白芷混合药液,其中白芷与细辛的比例为33:17。2.采用琼脂稀释法测定各药液的最低抑菌浓度。将P.gingivaaisAlTCC33277接种于含药琼脂平板上,37℃厌氧培养(80%N2、10%CO2、10%H2)72h,观察菌落形成情况。3.收集健康牙龈组织,采用改良组织块酶消化法进行人牙龈成纤维细胞原代培养,并通过免疫组化法鉴定细胞来源。4.以茵陈、野菊花、金银花及白芷+细辛为因素,以对牙龈卟啉单胞菌抑菌活性、生物膜形成抑制率及对人牙龈成纤维细胞毒性为考察指标,通过均匀设计法,筛选出最优配比,并以此配比制备复方金银花中药漱口水原液。5.将漱口水原液作用于大鼠口腔内,每天3次,每次40μL。阴性对照组为灭菌水。每天观察大鼠大体表现及记录体重。作用4周后,处死大鼠,取大鼠肝、脾、肺、肾组织,制作HE染色切片进行组织学观察。6.将30只大鼠随机分为药物作用组(T组)、阴性对照组(N组)及牙周炎造模组(P组)三组,采用丝线结扎+涂布牙龈卟啉单胞菌+高糖饮食法构建大鼠实验性牙周炎模型,造模期间观察大鼠口腔内牙周情况,第4周时从各组中随机选择处死大鼠,制作HE染色切片观察造模情况。7.将漱口水原液涂布于T组大鼠左侧上颌第二磨牙龈缘,每天3次,每次20μL。N组涂布灭菌水。P组不作任何处理。持续用药20 d后,处死所有大鼠,取造模牙及牙周组织制作HE染色切片进行观察。研究结果1.各单味药液对牙龈卟啉单胞菌最低有效抑菌浓度分别为:茵陈40 mg/mL,野菊花 10mg/mL,金银花 10mg/mL,白芷+细辛(33:17)40mg/mL。2.通过二项式逐步回归得出5种药物的最佳配比为:茵陈55 mg/mL,野菊花45 mg/mL,金银花 40 mg/mL,白芷+细辛(33:17)40mg/mL。3.毒性实验结果显示:实验组与阴性对照组大鼠体重无明显差异,大鼠肝、脾、肺、肾组织未见明显异常。4.成功构建大鼠实验性牙周炎模型。大鼠口腔检查及组织HE染色切片结果显示:与P组及N组相比,T组牙周组织炎症细胞浸润减少,牙周膜纤维排列整齐。研究结论1.通过均匀设计法筛选出的复方金银花中药漱口水无明显急性毒性作用,且短期内连续使用无毒性作用。2.复方金银花中药漱口水能有效减轻大鼠实验性牙周炎的牙龈红肿、探诊出血等炎症反应,促进牙周组织愈合。
[Abstract]:Background and objective periodontitis are the main causes of dental loss in adults in China. At present, most scholars believe that periodontitis is a multifactor disease, among which the bacteria and their toxic products are essential for the initiation of periodontitis. Therefore, control and removal of dental plaque is the key to the treatment of periodontitis. The common method of spot control is mechanical plaque control, including gingival cleaning, subgingival scaling, brushing and so on. On the basis of basic treatment, gargle with antibacterial ingredients can be used as an auxiliary means to achieve better control effect of plaque. Gargle is an important role in the prevention and adjuvant treatment of oral diseases. Different ingredients, mouthwash can be divided into caries proof gargle, anti sensitive gargle, antibiotic gargle, inhibition of dental plaque gargle, breath fresh gargle and herbal gargle. It has good effects in eliminating odors, dental caries, anti-inflammatory and other diseases, often as periodontitis, gingivitis, periodontitis and oral ulcers. Among them, the inhibition of dental plaque gargle as a means of controlling self plaque can enter the area of toothbrush, dental floss, cleaning device and other instruments which can not reach the toothbrush, dental floss and sanitary ware, and supplement the defects of mechanical plaque control. "Bing Tai", a new type of iodine gargle with iodine, which can cause tooth surface coloring, oral bacteria dissonance and oral mucosa irritation for a long time. Therefore, to find effective, safe, non alcohol and long term use of mouthwash is a new hot spot in the prevention and treatment of periodontitis. In China, the use of natural drugs has a long history, small toxic side effects, convenient extraction and economical benefits. Some of them are widely used in basic research and clinical practice. The use of traditional Chinese medicine is widely used in oral cavity. A large number of studies have shown that traditional Chinese medicine has a good effect on the treatment of periodontitis and oral ulcer. There are few studies, but the traditional Chinese medicine compound can reflect the characteristics of TCM syndrome differentiation, give full play to the synergism of the medicine and the multi target effect of various effective components of traditional Chinese medicine. With people's attention to the safety of mouthwash, the advantages of the Chinese medicine gargle have gradually emerged. The traditional Chinese medicine gargle, which has been sold in the market, is common, but it is common. It has the problems of turbid color, poor taste or alcohol, which makes its acceptance lower than that of other kinds of mouthwash. Moreover, there is less systematic study on the study of Chinese herbal gargle from basic to application, lack of scientific and objective observation, and less reports on its toxicity and adverse reaction, and can not provide experimental basis for clinical application. The theory of traditional Chinese medicine (TCM) holds that hot stomach heat, deficiency of kidney qi and deficiency of Qi and blood are the main causes of periodontitis. The bioactivity of Yin Chen, chrysanthemum chrysanthemum, honeysuckle, angelica dahuricae, asarum is strong, and has been proved to have anti-inflammatory, immune and other functions, and has inhibitory effect on various bacteria including oral pathogenic bacteria. It is not reported in the prevention and treatment of periodontitis. To sum up, this study is to extract single flavor liquid from Yinchen, chrysanthemum chrysanthemum and honeysuckle, and extract the mixture of Asarum and Angelica dahurica. Through uniform design, the best ratio of 5 drugs is obtained. The preparation of compound honeysuckle gargle, through zoological experiments, is used to study the toxicity of the mouthwash and to the periodontium. The efficacy of the disease provides a scientific basis for the further development of a safe and effective compound Chinese herbal gargle. 1. the extraction method of volatile oil extraction is used to extract 5 kinds of medicinal herbs: Yinchen, wild chrysanthemum, honeysuckle, asarum and dahurica dahurica. The preparation of the mixture of Yinchen, chrysanthemum, honeysuckle and Angelica dahurica is mixed with Radix Angelicae dahuricae and asarum The minimum inhibitory concentration of 33:17.2. was measured by agar dilution method. P.gingivaaisAlTCC33277 was inoculated on drug agar plate and anaerobic culture (80%N2,10%CO2,10%H2) 72h at 37 C. The formation of healthy gingival tissue was collected by.3.. The modified tissue block enzyme digestion was used to carry out the primary culture of human gingival fibroblasts. The cell source.4. was identified by immunohistochemistry. The inhibitory activity of Porphyromonas gingivalis, the inhibition rate of the biofilm formation and the toxicity of the gingival fibroblasts were investigated. The optimum ratio was screened by the uniform design method, and the compound gold and silver were prepared by this method. A mouthwash liquid.5. was used to treat the mouthwash liquid in the oral cavity of rats. 3 times a day, 40 L. negative control group was sterilized. Every day, the rats were observed and recorded the body weight. After 4 weeks, rats were killed and rat liver, spleen, lung, and kidney tissue were taken to make HE staining sections for histological observation, and 30 rats were randomly divided into drugs. Three groups (group T), negative control group (group N) and group of periodontitis (group P) were used to construct experimental periodontitis model in rats by ligation of silk thread plus porphyromoninomonas gingivalis + high sugar diet. During the modeling period, the periodontal condition of the rats was observed. At fourth weeks, the rats were randomly selected from each group, and the HE staining section was made to observe the mold making. Case.7. applied the mouthwash to the left upper maxillary second molar gingival margin of group T rats, 3 times a day, 20 mu L.N group was coated with sterilized water in group.P without any treatment. After sustained drug use, all rats were killed, the molars and periodontium were taken to make a HE staining section for observation. The results of the study were 1. of the most single flavour of porphyrinomonas gingivalis. The low effective bacteriostasis concentration is: 40 mg/mL of Yin Chen, chrysanthemum 10mg/mL, honeysuckle 10mg/mL and Radix Angelicae dahuricae + Asarum (33:17) 40mg/mL.2. through the binomial regression, the best ratio of 5 kinds of drugs is: Yin Chen 55 mg/mL, chrysanthemum 45 mg/mL, honeysuckle 40 mg/mL, Radix Angelicae dahuricae + thin symplecum (33:17) 40mg/mL.3. toxicity test results show: experimental group and negative The rats in the control group had no significant difference in weight, and the rat liver, spleen, lung and kidney tissue had no obvious abnormal.4. model of experimental periodontitis. The results of oral examination and tissue HE staining showed that compared with group P and N, the infiltration of inflammatory cells in the periodontal tissue of T group decreased and the periodontal ligament was arranged neatly. Conclusion 1. through uniformity of research conclusion The compound golden honeysuckle gargle had no obvious acute toxicity, and the continuous use of nontoxic effect of.2. gargle in the short term could effectively reduce the gingival redness and swelling of experimental periodontitis in rats, explore the bleeding and other inflammatory reactions, and promote the healing of the periodontal tissue.

【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781.42

【参考文献】