雙醋瑞因?qū)Υ笫髮嶒炐匝乐苎椎闹委熥饔?/H1>
發(fā)布時間：2018-06-30 16:39
本文選題：牙周炎 + 雙醋瑞因�。� 參考：《河北醫(yī)科大學(xué)》2014年碩士論文
【摘要】：目的：通過對雄性wistar大鼠牙周結(jié)扎的方法建立牙周炎動物模型，觀察雙醋瑞因?qū)ρ乐苎状笫蟮闹委熥饔�，為雙醋瑞因作為牙周炎的藥物治療提供實驗性依據(jù)。 方法：選用純種3月齡雄性wistar大鼠40只，隨機分成四組：正常組10只、牙周炎組10只、4周用藥組10只、8周用藥組10只。牙周炎、牙周炎用藥組在10%水合氯醛腹腔（330mg/kg）麻醉下，用直徑為0.2mm的正畸不銹鋼結(jié)扎絲雙側(cè)結(jié)扎上頜第一磨牙，將結(jié)扎絲放入游離齦內(nèi)，不能損傷牙齦結(jié)合上皮，結(jié)扎當(dāng)天，即用高糖水代替飲用水喂養(yǎng)。牙周炎造模一個月后，隨機抽取正常組和牙周炎組大鼠，處死，取其上頜骨，制作上頜第一磨牙牙體牙周聯(lián)合切片，組織學(xué)觀察，鑒定牙周炎模型。牙周炎模型鑒定成功后，用藥組用雙醋瑞因膠囊用2ml生理鹽水按100mg/kg配制成為混懸液，每天一次2ml灌胃；牙周炎則每天一次2ml生理鹽水灌胃。服藥4、8周后，隨機抽取大鼠各10只，股動脈放血處死，留全血備用，，取大鼠上頜骨，立即放入10%甲醛溶液中固定，24h后采用10%的EDTA溶液脫鈣60天，制作牙體牙周聯(lián)合組織切片，進(jìn)行HE染色，光鏡觀察；抗酒石酸酸性磷酸酶染色，觀察破骨細(xì)胞的分布及數(shù)量情況并作統(tǒng)計學(xué)分析處理；IL-1β免疫組織化學(xué)染色，觀察IL-1β的表達(dá)，對觀察結(jié)果進(jìn)行統(tǒng)計學(xué)分析。 結(jié)果: 1牙周結(jié)扎后，牙周炎組與用藥組大鼠食欲減退，精神狀態(tài)不佳，體重較正常組減輕；造模一周后，大鼠飲食逐漸恢復(fù)正常，精神狀態(tài)良好。牙周炎造模成功后，牙周炎組及用藥組開始分別灌胃生理鹽水、雙醋瑞因，用藥組大鼠出現(xiàn)輕度腹瀉的癥狀，尿量增加，尿液顏色偏黃，進(jìn)食較少，精神較萎靡，一周后腹瀉癥狀減輕，精神逐漸恢復(fù)正常。灌胃8周后，各組大鼠體重均有增加，正常組幅度最大，牙周炎組最小。 2造模4周后，取其上頜骨，肉眼觀察，正常組無明顯炎癥反應(yīng)，探診無出血;牙周炎組則表現(xiàn)為牙齦紅腫，探診出血明顯，牙齦與牙面不貼合，食物殘屑堆積，形成較淺的牙周袋。制作上頜第一磨牙牙體牙周聯(lián)合組織切片，顯示正常組牙周結(jié)構(gòu)完整，無炎癥反應(yīng)。牙周炎組牙周炎癥明顯，牙齦上皮內(nèi)有大量炎性細(xì)胞浸潤，結(jié)合上皮與牙根結(jié)合不緊密，向根方移位，牙周膜內(nèi)毛細(xì)血管擴張出血，牙槽嵴頂有少量吸收。說明牙周炎模型建立成功。 3大鼠牙周組織HE染色組織形態(tài)學(xué)觀察：正常組：結(jié)合上皮位于釉牙骨質(zhì)界處，無炎癥細(xì)胞浸潤，牙周膜纖維有序排列，牙槽嵴頂無明顯吸收。牙周炎組：結(jié)合上皮向根方移位，周圍有大量的炎性細(xì)胞浸潤，形成較深的牙周袋，毛細(xì)血管擴張出血，牙周膜間隙增寬，牙周膜纖維排列紊亂，牙槽嵴頂吸收明顯。4周用藥組：牙周袋變淺，上皮下方仍有炎性細(xì)胞浸潤，牙槽嵴頂見少量成骨細(xì)胞，牙槽嵴頂周圍見少量破骨細(xì)胞。8周用藥組：牙周袋變淺，牙槽嵴頂見大量成骨細(xì)胞，有新骨形成。 4抗酒石酸酸性磷酸酶染色法（TRAP）測定大鼠牙周組織中破骨細(xì)胞的分布和表達(dá)水平：正常組：牙槽嵴頂邊緣可見個別散在分布的破骨細(xì)胞。牙周炎組：牙槽嵴周圍見大量的破骨細(xì)胞密集分布。4周用藥組：破骨細(xì)胞仍少量分布于牙槽嵴頂周圍。8周用藥組：牙槽嵴頂周圍破骨細(xì)胞罕見。觀察牙槽嵴頂冠1/3區(qū)域破骨細(xì)胞分布并進(jìn)行統(tǒng)計分析，牙周炎組明顯高于正常組及用藥組(P0.01)，8周用藥組與正常組未見明顯差異（P0.05），4周用藥組高于8周用藥組（P0.01）。 5免疫組化測定IL-1β在大鼠牙槽骨中的分布及表達(dá)水平，IL-1β存在上皮細(xì)胞、中性粒細(xì)胞、單核-巨噬細(xì)胞、血管內(nèi)皮細(xì)胞、成纖維細(xì)胞等細(xì)胞胞漿及牙周膜基質(zhì)，正常組：陽性細(xì)胞少，牙周炎組：成纖維細(xì)胞的胞漿,炎性細(xì)胞灶呈強陽性表達(dá)，4周用藥組：成纖維細(xì)胞等呈陽性表達(dá)，8周用藥組：陽性細(xì)胞少，呈弱陽性表達(dá)。光密度值測定牙周炎對照組明顯高于正常組和用藥組（P0.01），正常組與8周用藥組之間無明顯差異（P0.05）,4周用藥組高于8周用藥組(P0.01)。 結(jié)論： 1通過結(jié)扎大鼠上頜第一磨牙成功地建立了大鼠牙周炎的模型。 2雙醋瑞因膠囊顯著降低了IL-1β的表達(dá)，減輕了牙周局部炎癥，抑制大鼠牙周組織中破骨細(xì)胞的形成，誘導(dǎo)成骨，促進(jìn)了牙槽骨新骨的形成，對牙周炎有治療作用。
[Abstract]:Objective: to establish an animal model of periodontitis by the method of periodontal ligature in male Wistar rats, and to observe the therapeutic effect of Diacetate on periodontitis rats, and provide experimental basis for the treatment of diacetate as a drug for periodontitis.
Methods: 40 purebred 3 month old male Wistar rats were randomly divided into four groups: 10 in the normal group, 10 in the periodontitis group, 10 in the 4 week drug group and 10 in the 8 week drug group. The group of periodontitis and periodontitis was anesthetized in 10% chloral abdominal cavity (330mg/kg), and the first molar of the upper jaw was ligated with a orthodontic stainless steel ligation with a diameter of 0.2mm. In the day of ligating the gingiva, the gingival combined epithelium can not be damaged, and the day of ligating the gingiva, that is, to replace the drinking water with high sugar water. After a month of periodontitis, the normal group and the periodontitis group were randomly selected and executed, and the upper jaw was taken, the periodontium section of the maxillary first molar was made, the periodontitis model was identified and the periodontitis model was identified. After the success of the type identification, the drug group used Diacerein Capsules to make a suspension of 100mg/kg with 2ml normal saline by using 2ml saline one time every day, while periodontitis was gavage with 2ml normal saline once a day. After taking the medicine for 4,8 weeks, 10 rats were randomly selected, the femoral artery was put to death, the whole blood was left, the maxilla of the rat was taken, and 10% Formaldehyde Solution was immediately put into the maxilla. After 24h, 10% EDTA solution was decalcified after 60 days, the joint tissue section of tooth body was made, HE staining, light microscope observation, anti tartaric acid acid phosphatase staining, observation of the distribution and quantity of osteoclast were observed, and IL-1 beta immunohistochemical staining was used to observe the expression of IL-1 beta, and the observation results were observed. Statistical analysis.
Result:
After 1 periodontal ligations, the appetite of the periodontitis group and the drug group was reduced, the mental state was poor and the weight was less than the normal group. After a week, the diet of rats gradually recovered to normal, and the mental state was good. After the success of the periodontitis model, the periodontitis group and the drug group began to irrigate the normal saline, double vinegar reiin, and the rats in the drug group had mild diarrhea. After 8 weeks of gavage, the weight of the rats in each group increased, the normal group was the largest, and the periodontitis group was the smallest.
2 after 4 weeks, the upper jaw and the naked eye were observed with the naked eye. There was no obvious inflammatory reaction in the normal group and no bleeding was found in the normal group. The periodontitis group was characterized by red and swollen gums, the bleeding of the gums and the teeth, the accumulation of food debris and the formation of a shallow periodontal pocket. The joint tissue section of the first molar teeth of the upper jaw was made to show the normal periodontal structure of the group. There were obvious periodontitis in the periodontitis group, a large number of inflammatory cells infiltrated in the gingival epithelium, the combination of the epithelium and the root of the teeth was not close, the root was shifted to the root, the blood capillary in the periodontal membrane was dilated and bleed, and the crest of the alveolar ridge had a small amount of absorption. It was a successful model of periodontitis.
3 rat periodontal tissue HE staining histomorphology observation: the normal group: the normal group: the epithelium is located at the enamel cementum boundary, no inflammatory cell infiltration, the periodontal ligament is arranged in order, the alveolar crest has no obvious absorption. The periodontitis group is shifted to the root side, and there are a large number of inflammatory cells infiltrating around, forming a deeper periodontal pocket and capillary expansion. The gap between the periodontal membrane and the periodontal ligament was disordered and the alveolar crest was absorbed by.4 weeks. The periodontal pocket became shallow, the alveolar ridge still had inflammatory cell infiltration, a small amount of osteoblasts were found at the top of the alveolar ridge, and a small amount of osteoclasts around the alveolar crest were seen around the top of the alveolar ridge for.8 weeks: the periodontal pocket became shallow and a large number of osteoblasts were found at the top of the alveolar ridge. There were a large number of osteoblasts. New bone formation.
4 anti tartaric acid acid phosphatase staining (TRAP) was used to determine the distribution and expression of osteoclast in the periodontal tissue of rats: in the normal group, a few scattered osteoclasts were found on the top of the alveolar ridge. In the periodontitis group, a large number of osteoclasts were distributed around the alveolar ridge for.4 weeks, and the osteoclasts were still small in the alveolus. .8 weeks around the crest: osteoclasts around the alveolar crest were rare. The distribution of osteoclasts in the 1/3 region of the alveolar crest was observed and statistically analyzed. The periodontitis group was significantly higher than the normal group and the drug group (P0.01). There was no significant difference between the 8 week group and the normal group (P0.05), and the 4 week medication group was higher than the 8 week medication group (P0.01).
5 immunohistochemistry was used to determine the distribution and expression level of IL-1 beta in the alveolar bone of rats. IL-1 beta had epithelial cells, neutrophils, mononuclear macrophages, vascular endothelial cells, fibroblasts, and periodontal membrane matrix. The normal group was few positive cells, the periodontitis group was the cytoplasm of fibroblasts, and the inflammatory cytoplasm showed a strong positive table. 4 weeks of medication group: fibroblast positive expression, 8 weeks of drug group: the positive cells were less, showed weak positive expression. The light density value of periodontitis control group was significantly higher than the normal group and the drug group (P0.01), there was no significant difference between the normal group and the 8 week medication group (P0.05), 4 weeks medication group was higher than the 8 week medication group (P0.01).
Conclusion:
1 a rat periodontitis model was successfully established by ligating the maxillary first molar in rats.
2 Diacerein Capsules significantly reduced the expression of IL-1 beta, alleviated local inflammation in periodontitis, inhibited the formation of osteoclasts in the periodontal tissue of rats, induced osteogenesis, promoted the formation of new bone in the alveolar bone, and had a therapeutic effect on periodontitis.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R781.4
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本文選題：牙周炎 + 雙醋瑞因�。� 參考：《河北醫(yī)科大學(xué)》2014年碩士論文
【摘要】：目的：通過對雄性wistar大鼠牙周結(jié)扎的方法建立牙周炎動物模型，觀察雙醋瑞因?qū)ρ乐苎状笫蟮闹委熥饔�，為雙醋瑞因作為牙周炎的藥物治療提供實驗性依據(jù)。 方法：選用純種3月齡雄性wistar大鼠40只，隨機分成四組：正常組10只、牙周炎組10只、4周用藥組10只、8周用藥組10只。牙周炎、牙周炎用藥組在10%水合氯醛腹腔（330mg/kg）麻醉下，用直徑為0.2mm的正畸不銹鋼結(jié)扎絲雙側(cè)結(jié)扎上頜第一磨牙，將結(jié)扎絲放入游離齦內(nèi)，不能損傷牙齦結(jié)合上皮，結(jié)扎當(dāng)天，即用高糖水代替飲用水喂養(yǎng)。牙周炎造模一個月后，隨機抽取正常組和牙周炎組大鼠，處死，取其上頜骨，制作上頜第一磨牙牙體牙周聯(lián)合切片，組織學(xué)觀察，鑒定牙周炎模型。牙周炎模型鑒定成功后，用藥組用雙醋瑞因膠囊用2ml生理鹽水按100mg/kg配制成為混懸液，每天一次2ml灌胃；牙周炎則每天一次2ml生理鹽水灌胃。服藥4、8周后，隨機抽取大鼠各10只，股動脈放血處死，留全血備用，，取大鼠上頜骨，立即放入10%甲醛溶液中固定，24h后采用10%的EDTA溶液脫鈣60天，制作牙體牙周聯(lián)合組織切片，進(jìn)行HE染色，光鏡觀察；抗酒石酸酸性磷酸酶染色，觀察破骨細(xì)胞的分布及數(shù)量情況并作統(tǒng)計學(xué)分析處理；IL-1β免疫組織化學(xué)染色，觀察IL-1β的表達(dá)，對觀察結(jié)果進(jìn)行統(tǒng)計學(xué)分析。 結(jié)果: 1牙周結(jié)扎后，牙周炎組與用藥組大鼠食欲減退，精神狀態(tài)不佳，體重較正常組減輕；造模一周后，大鼠飲食逐漸恢復(fù)正常，精神狀態(tài)良好。牙周炎造模成功后，牙周炎組及用藥組開始分別灌胃生理鹽水、雙醋瑞因，用藥組大鼠出現(xiàn)輕度腹瀉的癥狀，尿量增加，尿液顏色偏黃，進(jìn)食較少，精神較萎靡，一周后腹瀉癥狀減輕，精神逐漸恢復(fù)正常。灌胃8周后，各組大鼠體重均有增加，正常組幅度最大，牙周炎組最小。 2造模4周后，取其上頜骨，肉眼觀察，正常組無明顯炎癥反應(yīng)，探診無出血;牙周炎組則表現(xiàn)為牙齦紅腫，探診出血明顯，牙齦與牙面不貼合，食物殘屑堆積，形成較淺的牙周袋。制作上頜第一磨牙牙體牙周聯(lián)合組織切片，顯示正常組牙周結(jié)構(gòu)完整，無炎癥反應(yīng)。牙周炎組牙周炎癥明顯，牙齦上皮內(nèi)有大量炎性細(xì)胞浸潤，結(jié)合上皮與牙根結(jié)合不緊密，向根方移位，牙周膜內(nèi)毛細(xì)血管擴張出血，牙槽嵴頂有少量吸收。說明牙周炎模型建立成功。 3大鼠牙周組織HE染色組織形態(tài)學(xué)觀察：正常組：結(jié)合上皮位于釉牙骨質(zhì)界處，無炎癥細(xì)胞浸潤，牙周膜纖維有序排列，牙槽嵴頂無明顯吸收。牙周炎組：結(jié)合上皮向根方移位，周圍有大量的炎性細(xì)胞浸潤，形成較深的牙周袋，毛細(xì)血管擴張出血，牙周膜間隙增寬，牙周膜纖維排列紊亂，牙槽嵴頂吸收明顯。4周用藥組：牙周袋變淺，上皮下方仍有炎性細(xì)胞浸潤，牙槽嵴頂見少量成骨細(xì)胞，牙槽嵴頂周圍見少量破骨細(xì)胞。8周用藥組：牙周袋變淺，牙槽嵴頂見大量成骨細(xì)胞，有新骨形成。 4抗酒石酸酸性磷酸酶染色法（TRAP）測定大鼠牙周組織中破骨細(xì)胞的分布和表達(dá)水平：正常組：牙槽嵴頂邊緣可見個別散在分布的破骨細(xì)胞。牙周炎組：牙槽嵴周圍見大量的破骨細(xì)胞密集分布。4周用藥組：破骨細(xì)胞仍少量分布于牙槽嵴頂周圍。8周用藥組：牙槽嵴頂周圍破骨細(xì)胞罕見。觀察牙槽嵴頂冠1/3區(qū)域破骨細(xì)胞分布并進(jìn)行統(tǒng)計分析，牙周炎組明顯高于正常組及用藥組(P0.01)，8周用藥組與正常組未見明顯差異（P0.05），4周用藥組高于8周用藥組（P0.01）。 5免疫組化測定IL-1β在大鼠牙槽骨中的分布及表達(dá)水平，IL-1β存在上皮細(xì)胞、中性粒細(xì)胞、單核-巨噬細(xì)胞、血管內(nèi)皮細(xì)胞、成纖維細(xì)胞等細(xì)胞胞漿及牙周膜基質(zhì)，正常組：陽性細(xì)胞少，牙周炎組：成纖維細(xì)胞的胞漿,炎性細(xì)胞灶呈強陽性表達(dá)，4周用藥組：成纖維細(xì)胞等呈陽性表達(dá)，8周用藥組：陽性細(xì)胞少，呈弱陽性表達(dá)。光密度值測定牙周炎對照組明顯高于正常組和用藥組（P0.01），正常組與8周用藥組之間無明顯差異（P0.05）,4周用藥組高于8周用藥組(P0.01)。 結(jié)論： 1通過結(jié)扎大鼠上頜第一磨牙成功地建立了大鼠牙周炎的模型。 2雙醋瑞因膠囊顯著降低了IL-1β的表達(dá)，減輕了牙周局部炎癥，抑制大鼠牙周組織中破骨細(xì)胞的形成，誘導(dǎo)成骨，促進(jìn)了牙槽骨新骨的形成，對牙周炎有治療作用。
[Abstract]:Objective: to establish an animal model of periodontitis by the method of periodontal ligature in male Wistar rats, and to observe the therapeutic effect of Diacetate on periodontitis rats, and provide experimental basis for the treatment of diacetate as a drug for periodontitis.
Methods: 40 purebred 3 month old male Wistar rats were randomly divided into four groups: 10 in the normal group, 10 in the periodontitis group, 10 in the 4 week drug group and 10 in the 8 week drug group. The group of periodontitis and periodontitis was anesthetized in 10% chloral abdominal cavity (330mg/kg), and the first molar of the upper jaw was ligated with a orthodontic stainless steel ligation with a diameter of 0.2mm. In the day of ligating the gingiva, the gingival combined epithelium can not be damaged, and the day of ligating the gingiva, that is, to replace the drinking water with high sugar water. After a month of periodontitis, the normal group and the periodontitis group were randomly selected and executed, and the upper jaw was taken, the periodontium section of the maxillary first molar was made, the periodontitis model was identified and the periodontitis model was identified. After the success of the type identification, the drug group used Diacerein Capsules to make a suspension of 100mg/kg with 2ml normal saline by using 2ml saline one time every day, while periodontitis was gavage with 2ml normal saline once a day. After taking the medicine for 4,8 weeks, 10 rats were randomly selected, the femoral artery was put to death, the whole blood was left, the maxilla of the rat was taken, and 10% Formaldehyde Solution was immediately put into the maxilla. After 24h, 10% EDTA solution was decalcified after 60 days, the joint tissue section of tooth body was made, HE staining, light microscope observation, anti tartaric acid acid phosphatase staining, observation of the distribution and quantity of osteoclast were observed, and IL-1 beta immunohistochemical staining was used to observe the expression of IL-1 beta, and the observation results were observed. Statistical analysis.
Result:
After 1 periodontal ligations, the appetite of the periodontitis group and the drug group was reduced, the mental state was poor and the weight was less than the normal group. After a week, the diet of rats gradually recovered to normal, and the mental state was good. After the success of the periodontitis model, the periodontitis group and the drug group began to irrigate the normal saline, double vinegar reiin, and the rats in the drug group had mild diarrhea. After 8 weeks of gavage, the weight of the rats in each group increased, the normal group was the largest, and the periodontitis group was the smallest.
2 after 4 weeks, the upper jaw and the naked eye were observed with the naked eye. There was no obvious inflammatory reaction in the normal group and no bleeding was found in the normal group. The periodontitis group was characterized by red and swollen gums, the bleeding of the gums and the teeth, the accumulation of food debris and the formation of a shallow periodontal pocket. The joint tissue section of the first molar teeth of the upper jaw was made to show the normal periodontal structure of the group. There were obvious periodontitis in the periodontitis group, a large number of inflammatory cells infiltrated in the gingival epithelium, the combination of the epithelium and the root of the teeth was not close, the root was shifted to the root, the blood capillary in the periodontal membrane was dilated and bleed, and the crest of the alveolar ridge had a small amount of absorption. It was a successful model of periodontitis.
3 rat periodontal tissue HE staining histomorphology observation: the normal group: the normal group: the epithelium is located at the enamel cementum boundary, no inflammatory cell infiltration, the periodontal ligament is arranged in order, the alveolar crest has no obvious absorption. The periodontitis group is shifted to the root side, and there are a large number of inflammatory cells infiltrating around, forming a deeper periodontal pocket and capillary expansion. The gap between the periodontal membrane and the periodontal ligament was disordered and the alveolar crest was absorbed by.4 weeks. The periodontal pocket became shallow, the alveolar ridge still had inflammatory cell infiltration, a small amount of osteoblasts were found at the top of the alveolar ridge, and a small amount of osteoclasts around the alveolar crest were seen around the top of the alveolar ridge for.8 weeks: the periodontal pocket became shallow and a large number of osteoblasts were found at the top of the alveolar ridge. There were a large number of osteoblasts. New bone formation.
4 anti tartaric acid acid phosphatase staining (TRAP) was used to determine the distribution and expression of osteoclast in the periodontal tissue of rats: in the normal group, a few scattered osteoclasts were found on the top of the alveolar ridge. In the periodontitis group, a large number of osteoclasts were distributed around the alveolar ridge for.4 weeks, and the osteoclasts were still small in the alveolus. .8 weeks around the crest: osteoclasts around the alveolar crest were rare. The distribution of osteoclasts in the 1/3 region of the alveolar crest was observed and statistically analyzed. The periodontitis group was significantly higher than the normal group and the drug group (P0.01). There was no significant difference between the 8 week group and the normal group (P0.05), and the 4 week medication group was higher than the 8 week medication group (P0.01).
5 immunohistochemistry was used to determine the distribution and expression level of IL-1 beta in the alveolar bone of rats. IL-1 beta had epithelial cells, neutrophils, mononuclear macrophages, vascular endothelial cells, fibroblasts, and periodontal membrane matrix. The normal group was few positive cells, the periodontitis group was the cytoplasm of fibroblasts, and the inflammatory cytoplasm showed a strong positive table. 4 weeks of medication group: fibroblast positive expression, 8 weeks of drug group: the positive cells were less, showed weak positive expression. The light density value of periodontitis control group was significantly higher than the normal group and the drug group (P0.01), there was no significant difference between the normal group and the 8 week medication group (P0.05), 4 weeks medication group was higher than the 8 week medication group (P0.01).
Conclusion:
1 a rat periodontitis model was successfully established by ligating the maxillary first molar in rats.
2 Diacerein Capsules significantly reduced the expression of IL-1 beta, alleviated local inflammation in periodontitis, inhibited the formation of osteoclasts in the periodontal tissue of rats, induced osteogenesis, promoted the formation of new bone in the alveolar bone, and had a therapeutic effect on periodontitis.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R781.4
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