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研究生:蔡凰慈
研究生(外文):Tsai, Huang-Tzu
論文名稱:丹參對大鼠實驗性牙周炎之改善效果
論文名稱(外文):Ameliorative Effect Of Danshen On The Experimental Periodontitis In Rats
指導教授:傅鍔傅鍔引用關係
指導教授(外文):Fu, Earl
口試委員:聶鑫江正陽沈一慶張溫良涂筱培
口試委員(外文):Nieh, ShieChiang, Cheng-YangShen, E-ChinChang, Wen-LiangTu, Shiao-Pei
口試日期:2012-05-31
學位類別:碩士
校院名稱:國防醫學院
系所名稱:牙醫科學研究所
學門:醫藥衛生學門
學類:牙醫學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:60
中文關鍵詞:丹參大鼠實驗性牙周病發炎
外文關鍵詞:danshenratexperimental periodontitisinflammationSalvia miltiorrhiza
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背景:丹參特指唇形科植物Salvia miltiorrhiza Bunge (鼠尾草屬) 之乾燥根莖部分,為數百年來中國傳統醫學中普遍且常用的中藥材之一,研究發現丹參抽取物具有多重之藥理、生理作用,臨床上更是被廣泛地使用在治療多種慢性病、尤以心血管疾病為主;研究亦證實丹參具有抗發炎之效果,不論是活體外或活體內的實驗,皆能有效抑制發炎物質產生。
實驗目的:利用脂多醣注射牙齦組織、或以絲線圈結牙齒之兩種不同的大鼠實驗性牙周炎模型,觀察丹參對其所造成之齒槽骨吸收破壞的現象有何影響,並探討丹參抗牙周炎的效果。
材料與方法:實驗動物為6~7週大之Spraque-Dawley (S.D.) 雄性大白鼠;投藥時間為促發牙周炎的前一天起、每天給予一次,至實驗結束共八天為止,並分別以兩種方式誘發牙周炎:一、大鼠分為三組;將磷酸緩衝溶液 (PBS) 注射於其上顎左右兩側大臼齒間的顎側牙齦組織,以做為控制組,其餘兩組注射細菌之脂多醣 (LPS,5 μg/ml),且實驗組前一天即開始腹腔注射施打丹參酒精萃取液300 mg/kg;第八天犧牲所有動物,以酒石酸具耐受性酸性磷酸酵素 (TRAP) 染色,檢視牙齦組織中的蝕骨細胞表現量。二、大鼠分六組,除控制組外,以黑絲線將大鼠左右兩側下顎第一大臼齒、上顎第二大臼齒分別圈綁線結;實驗組前一天開始即給予丹參,劑量及投藥方式則分為:口服市售丹參單味中藥粉末200 mg/kg、500 mg/kg,或是腹腔注射丹參酒精萃取液100 mg/kg、300 mg/kg;所有動物於第八天犧牲後,利用牙科數位影像、微電腦斷層掃描與切片染色..等方法進行量測,觀察牙齦軟硬組織之之破壞量。
實驗結果: 蝕骨細胞數量在脂多醣注射後明顯增加,若合併給予丹參,則蝕骨細胞數量明顯較少;於綁線-誘發牙周炎之實驗,在數位X光片、微電腦斷層掃描或組織切片中,均可發現大鼠齒槽骨有明顯喪失,而若給予丹參,其骨頭破壞的現象則可獲得改善,組織學下發炎的結締組織與浸潤細胞也較不明顯。
結論:綜合本實驗結果,證實中藥丹參確具有減緩大鼠因脂多醣-誘發與綁線-誘發牙周炎後之牙周組織破壞現象,而具有作為緩解牙周炎、甚或治療其他發炎疾病之開發潛力;惟因其有效成分較複雜且多樣化,且其吸收有效濃度會隨投藥途徑不同而有所改變,故此部分未來仍尚待進一步探討。

Background: Danshen, the dried root of Salvia miltiorrhiza Bunge, is one of the most popular and commonly used traditional Chinese medicine (TCM) that have been used for thousands of years. The use of danshen has been mostly in the treatment of cardiovascular diseases, but its anti-inflammatory effect on dental aspect has never been explored and investigated yet. The purpose of this study is to examine the effect of danshen on different models of experimental periodontitis in rats by several approaches for assessing periodontal destruction.
Materials and methods:Maxillary palatal alveoli of the fifteen Spraque-Dawley (S.D.) rats were injected with daily lipopolysaccharide (LPS) injections (5 μg/mL) for 3 days. Danshen was given each day by intra-peritoneal injection (i.p.) 300 mg/kg on rats from the day before LPS injection. Five days later after finishing LPS injection, the animals were sacrificed and the osteoclasts on bony surfaces were counted under tartrate-resistant acid phosphatase staining. Moreover, the other thirty-six rats were divided in the following six groups:control, ligation and ligation plus danshen (oral administration 200 mg/kg and 500 mg/kg; i.p. 100 mg/kg and 300 mg/kg) given each day before the day of ligation. The effect of danshen on the ligature-induced periodontitis around maxillary second and mandibular first molars was evaluated by histometric, digital radiographic and micro-computed tomographic analysis on day 8.
Results:Osteoclasts occurred and increased after LPS injections. However, the groups dosed with danshen were found less osteoclasts formation. On the other hand, by using dental digital radiography and micro-computed tomography, the alveolar bone crest levels at ligation sites were found to be significantly more apical than that at the control group on days 8. However, the apically located bone crests rebounded in rats from the danshen-plus-ligation groups. Histometry demonstrated significantly more coronal alveolar crest bone levels, less inflammatory cells infiltration and less connective tissue attachments loss in experimental groups compared with those in the ligation group.
Conclusion:Danshen could reduce both LPS-induced and ligature-induced periodontal inflammatory response and tissue breakdown in rats. By histometric, radiographic and micro-computed tomographic analysis, we suggest that danshen may have an ameliorative effect on periodontal destruction.

第一章 緒言................................................................................................................1
第一節 牙周組織與牙周病之分類................................................................................................1
第二節 牙周病的致病機轉.....................................................................................................3
第三節 丹參及其成分.........................................................................................................6
第四節 研究目的............................................................................................................8
第二章 材料與方法...........................................................................................................9
第一節 主要儀器與藥品試劑...................................................................................................10
第二節 主要實驗方法與步驟...................................................................................................12
第三章 結果...............................................................................................................27
第一節 脂多醣-誘發實驗性牙周炎模式...........................................................................................28
第二節 結紮線-誘發實驗性牙周炎模式...........................................................................................30
第四章 討論...............................................................................................................40
第一節 丹參之相關探討......................................................................................................41
第二節 丹參作用於脂多醣-誘發實驗性牙周炎.....................................................................................45
第三節 丹參作用於結紮線-誘發實驗性牙周炎.....................................................................................46
第五章 結論...............................................................................................................48
第六章 參考文獻...........................................................................................................50
附錄......................................................................................................................54

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