双醋瑞因对大鼠实验性牙周炎的治疗作用

发布时间：2018-06-30 16:39

本文选题：牙周炎 + 双醋瑞因　；参考：《河北医科大学》2014年硕士论文

【摘要】：目的：通过对雄性wistar大鼠牙周结扎的方法建立牙周炎动物模型，观察双醋瑞因对牙周炎大鼠的治疗作用，为双醋瑞因作为牙周炎的药物治疗提供实验性依据。方法：选用纯种3月龄雄性wistar大鼠40只，随机分成四组：正常组10只、牙周炎组10只、4周用药组10只、8周用药组10只。牙周炎、牙周炎用药组在10%水合氯醛腹腔（330mg/kg）麻醉下，用直径为0.2mm的正畸不锈钢结扎丝双侧结扎上颌第一磨牙，将结扎丝放入游离龈内，不能损伤牙龈结合上皮，结扎当天，即用高糖水代替饮用水喂养。牙周炎造模一个月后，随机抽取正常组和牙周炎组大鼠，处死，取其上颌骨，制作上颌第一磨牙牙体牙周联合切片，组织学观察，鉴定牙周炎模型。牙周炎模型鉴定成功后，用药组用双醋瑞因胶囊用2ml生理盐水按100mg/kg配制成为混悬液，每天一次2ml灌胃；牙周炎则每天一次2ml生理盐水灌胃。服药4、8周后，随机抽取大鼠各10只，股动脉放血处死，留全血备用，，取大鼠上颌骨，立即放入10%甲醛溶液中固定，24h后采用10%的EDTA溶液脱钙60天，制作牙体牙周联合组织切片，进行HE染色，光镜观察；抗酒石酸酸性磷酸酶染色，观察破骨细胞的分布及数量情况并作统计学分析处理；IL-1β免疫组织化学染色，观察IL-1β的表达，对观察结果进行统计学分析。结果: 1牙周结扎后，牙周炎组与用药组大鼠食欲减退，精神状态不佳，体重较正常组减轻；造模一周后，大鼠饮食逐渐恢复正常，精神状态良好。牙周炎造模成功后，牙周炎组及用药组开始分别灌胃生理盐水、双醋瑞因，用药组大鼠出现轻度腹泻的症状，尿量增加，尿液颜色偏黄，进食较少，精神较萎靡，一周后腹泻症状减轻，精神逐渐恢复正常。灌胃8周后，各组大鼠体重均有增加，正常组幅度最大，牙周炎组最小。 2造模4周后，取其上颌骨，肉眼观察，正常组无明显炎症反应，探诊无出血;牙周炎组则表现为牙龈红肿，探诊出血明显，牙龈与牙面不贴合，食物残屑堆积，形成较浅的牙周袋。制作上颌第一磨牙牙体牙周联合组织切片，显示正常组牙周结构完整，无炎症反应。牙周炎组牙周炎症明显，牙龈上皮内有大量炎性细胞浸润，结合上皮与牙根结合不紧密，向根方移位，牙周膜内毛细血管扩张出血，牙槽嵴顶有少量吸收。说明牙周炎模型建立成功。 3大鼠牙周组织HE染色组织形态学观察：正常组：结合上皮位于釉牙骨质界处，无炎症细胞浸润，牙周膜纤维有序排列，牙槽嵴顶无明显吸收。牙周炎组：结合上皮向根方移位，周围有大量的炎性细胞浸润，形成较深的牙周袋，毛细血管扩张出血，牙周膜间隙增宽，牙周膜纤维排列紊乱，牙槽嵴顶吸收明显。4周用药组：牙周袋变浅，上皮下方仍有炎性细胞浸润，牙槽嵴顶见少量成骨细胞，牙槽嵴顶周围见少量破骨细胞。8周用药组：牙周袋变浅，牙槽嵴顶见大量成骨细胞，有新骨形成。 4抗酒石酸酸性磷酸酶染色法（TRAP）测定大鼠牙周组织中破骨细胞的分布和表达水平：正常组：牙槽嵴顶边缘可见个别散在分布的破骨细胞。牙周炎组：牙槽嵴周围见大量的破骨细胞密集分布。4周用药组：破骨细胞仍少量分布于牙槽嵴顶周围。8周用药组：牙槽嵴顶周围破骨细胞罕见。观察牙槽嵴顶冠1/3区域破骨细胞分布并进行统计分析，牙周炎组明显高于正常组及用药组(P0.01)，8周用药组与正常组未见明显差异（P0.05），4周用药组高于8周用药组（P0.01）。 5免疫组化测定IL-1β在大鼠牙槽骨中的分布及表达水平，IL-1β存在上皮细胞、中性粒细胞、单核-巨噬细胞、血管内皮细胞、成纤维细胞等细胞胞浆及牙周膜基质，正常组：阳性细胞少，牙周炎组：成纤维细胞的胞浆,炎性细胞灶呈强阳性表达，4周用药组：成纤维细胞等呈阳性表达，8周用药组：阳性细胞少，呈弱阳性表达。光密度值测定牙周炎对照组明显高于正常组和用药组（P0.01），正常组与8周用药组之间无明显差异（P0.05）,4周用药组高于8周用药组(P0.01)。结论： 1通过结扎大鼠上颌第一磨牙成功地建立了大鼠牙周炎的模型。 2双醋瑞因胶囊显著降低了IL-1β的表达，减轻了牙周局部炎症，抑制大鼠牙周组织中破骨细胞的形成，诱导成骨，促进了牙槽骨新骨的形成，对牙周炎有治疗作用。
[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.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.4

【参考文献】