本文選題：牙周炎 + 漱口水��； 參考：《南方醫(yī)科大學》2017年碩士論文

【摘要】：研究背景及目的牙周炎是造成我國成年人牙齒喪失的主要原因。目前,大多數(shù)學者認為,牙周炎是多因素疾病,其中牙菌斑生物膜的細菌及其毒性產物是引發(fā)牙周炎必不可少的始動因子。因此,控制和去除牙菌斑是牙周炎治療的關鍵。菌斑控制的常用方法為機械性菌斑控制法,包括齦上潔治、齦下刮治、刷牙等。在基礎治療的基礎上,以含有抗菌成分的漱口水作為輔助手段,能取得更好的菌斑控制效果。漱口水在口腔疾病的預防及輔助治療中起著重要的作用。根據(jù)功能及成分的不同,漱口水可分為防齲類漱口水、抗敏感類漱口水、抗生素類漱口水、抑制牙菌斑類漱口水、口氣清新類漱口水及中草藥類漱口水,在清除口腔異味、防齲齒、抗炎等方面有很好的效果,常作為牙周炎、牙齦炎、冠周炎及口腔潰瘍等疾病的輔助用藥。其中,抑制牙菌斑類漱口水作為自我菌斑控制手段的一種,可通過液體流動力進入牙刷、牙線、潔治器等器械不能到達的區(qū)域,補充機械性菌斑控制法的不足。目前,臨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,長期使用易引起牙面著色、口腔菌群失調、口腔黏膜刺激等問題。因此,尋找有效、安全、無酒精、可長期使用的漱口水成為牙周病防治的新熱點。在我國,天然藥物的應用歷史悠久、且毒副作用小,取材方便,經(jīng)濟實惠,有的還具有殺菌消炎作用,因而廣泛用于基礎研究及臨床實踐。中藥制劑在口腔中的運用廣泛,大量研究表明中藥在治療牙周病、口腔潰瘍等疾病上具有良好的療效。但目前對于該類中藥的報道多為某種單一中草藥的作用,對中藥復方的研究較少,而中藥復方更能體現(xiàn)中醫(yī)辨證論治的特點,充分發(fā)揮藥物的協(xié)同作用及中醫(yī)藥多種有效成分多靶點作用。隨著人們對漱口水安全性的關注,中藥類漱口水的優(yōu)勢逐漸顯現(xiàn)出來。市面上已有成品銷售的中藥類漱口水,但普遍存在色澤渾濁、口感較差或含有酒精等問題,使得其接受度較其他類漱口水低。且對于中草藥類漱口水的研究,從基礎到應用的系統(tǒng)研究較少,缺乏科學客觀的療效觀察,且對于其毒性及不良反應報道較少,未能給臨床應用提供實驗依據(jù)。中醫(yī)理論認為胃火熾熱、腎氣虛虧和氣血不足是牙周炎的主要病因。茵陳、野菊花、金銀花、白芷、細辛生物活性強,被證實具有抗炎、調節(jié)免疫等作用,且對包括口腔致病菌在內的多種細菌具有抑制作用。將這5種藥物進行復方配伍,應用于防治牙周疾病中尚未見報道。綜上所述,本研究擬從茵陳、野菊花、金銀花中提取單味藥液,并提取細辛+白芷混合藥液,通過均勻設計法獲得5種藥物的最佳配比,制備復方金銀花中藥漱口水,通過動物學實驗,研究該漱口水的毒性及對牙周疾病的功效,為進一步開發(fā)出安全有效的復方金銀花中藥漱口水提供科學依據(jù)。研究方法1.采用揮發(fā)油提取法對茵陳、野菊花、金銀花、細辛、白芷5種藥物進行提取,制備茵陳、野菊花、金銀花單味藥液及細辛+白芷混合藥液,其中白芷與細辛的比例為33:17。2.采用瓊脂稀釋法測定各藥液的最低抑菌濃度。將P.gingivaaisAlTCC33277接種于含藥瓊脂平板上,37℃厭氧培養(yǎng)(80%N2、10%CO2、10%H2)72h,觀察菌落形成情況。3.收集健康牙齦組織,采用改良組織塊酶消化法進行人牙齦成纖維細胞原代培養(yǎng),并通過免疫組化法鑒定細胞來源。4.以茵陳、野菊花、金銀花及白芷+細辛為因素,以對牙齦卟啉單胞菌抑菌活性、生物膜形成抑制率及對人牙齦成纖維細胞毒性為考察指標,通過均勻設計法,篩選出最優(yōu)配比,并以此配比制備復方金銀花中藥漱口水原液。5.將漱口水原液作用于大鼠口腔內,每天3次,每次40μL。陰性對照組為滅菌水。每天觀察大鼠大體表現(xiàn)及記錄體重。作用4周后,處死大鼠,取大鼠肝、脾、肺、腎組織,制作HE染色切片進行組織學觀察。6.將30只大鼠隨機分為藥物作用組(T組)、陰性對照組(N組)及牙周炎造模組(P組)三組,采用絲線結扎+涂布牙齦卟啉單胞菌+高糖飲食法構建大鼠實驗性牙周炎模型,造模期間觀察大鼠口腔內牙周情況,第4周時從各組中隨機選擇處死大鼠,制作HE染色切片觀察造模情況。7.將漱口水原液涂布于T組大鼠左側上頜第二磨牙齦緣,每天3次,每次20μL。N組涂布滅菌水。P組不作任何處理。持續(xù)用藥20 d后,處死所有大鼠,取造模牙及牙周組織制作HE染色切片進行觀察。研究結果1.各單味藥液對牙齦卟啉單胞菌最低有效抑菌濃度分別為:茵陳40 mg/mL,野菊花 10mg/mL,金銀花 10mg/mL,白芷+細辛(33:17)40mg/mL。2.通過二項式逐步回歸得出5種藥物的最佳配比為:茵陳55 mg/mL,野菊花45 mg/mL,金銀花 40 mg/mL,白芷+細辛(33:17)40mg/mL。3.毒性實驗結果顯示:實驗組與陰性對照組大鼠體重無明顯差異,大鼠肝、脾、肺、腎組織未見明顯異常。4.成功構建大鼠實驗性牙周炎模型。大鼠口腔檢查及組織HE染色切片結果顯示:與P組及N組相比,T組牙周組織炎癥細胞浸潤減少,牙周膜纖維排列整齊。研究結論1.通過均勻設計法篩選出的復方金銀花中藥漱口水無明顯急性毒性作用,且短期內連續(xù)使用無毒性作用。2.復方金銀花中藥漱口水能有效減輕大鼠實驗性牙周炎的牙齦紅腫、探診出血等炎癥反應,促進牙周組織愈合。
[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.

【學位授予單位】：南方醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R781.42

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