β-隐黄素对大鼠实验性牙周炎牙槽骨吸收的影响

发布时间：2018-05-07 10:58

本文选题：牙周炎 + 骨吸收　；参考：《郑州大学》2017年硕士论文

【摘要】：背景在牙周炎病变过程中,炎性细胞因子和化学因子的存在上调了核因子κB受体活化因子配体(Receptor activator of nuclear factor-kappa B ligand,RANKL)的表达,RANKL活化破骨细胞(Osteoclast,OC)前体细胞,随后成熟的多核OC吸附到矿化的骨基质表面进行骨吸收。牙槽骨吸收是导致牙齿松动和脱落的主要原因。预防牙槽骨的吸收,促进牙槽骨的修复和再生是治愈牙周炎的关键。β-隐黄素是一种发现于水果和蔬菜中的类胡萝卜素。β-隐黄素可以通过刺激成骨细胞(Osteoblast,OB)形成骨,抑制OC吸收骨来预防骨组织的丧失。以往的研究发现血清中低水平的β-隐黄素和牙周炎患病率增加有关,牙周炎患者血清中β-隐黄素水平明显较低;β-隐黄素可以抑制内毒素(Lipopolysaccharide,LPS)诱导的炎症反应,上调骨保护素(Osteoprotegerin,OPG)/RANKL的比值;注射β-隐黄素可以抑制LPS-诱导的小鼠牙槽骨骨密度(Bone mineral density,BMD)的降低。这些研究表明,β-隐黄素影响牙周炎的发生发展。然而,关于β-隐黄素对牙周炎牙槽骨吸收的影响研究较少,且其作用机理尚不十分明确。目的本研究的目的是通过检测牙槽骨丧失量(Alveolar bone loss,ABL)、OC数目、RANKL及OPG的表达来评估β-隐黄素对大鼠实验性牙周炎牙周组织的作用,检测其对牙周炎牙槽骨吸收的影响并分析其作用机制。方法30只雄性SD大鼠随机分为三组:1)正常对照组(N);2)牙周炎模型组(P);3)β-隐黄素干预组(E)。P组、E组进行实验性牙周炎造模,E组在造模的同时用β-隐黄素进行干预。实验性牙周炎模型的诱导:直径0.2mm的正畸结扎丝环绕在双侧上颌第二磨牙牙颈部,同时在颊侧牙龈龈沟内注射大肠杆菌LPS(30μl/鼠),每48h注射1次,共3次。作为对照,N组注射等量的生理盐水。β-隐黄素干预:E组在注射LPS后在相同位点注射β-隐黄素(12μl/鼠),每48h 1次,共3次。P组注射等量的玉米油代替。实验第8天大鼠腹腔注射过量的10%水合氯醛处死,取双侧上颌组织(含牙齿、牙龈、牙周膜、牙槽骨)。右侧上颌组织进行形态学分析,测量釉牙骨质界(Cemento-enamel junction,CEJ)到牙槽嵴顶(Alveolar bone crest,ABC)的距离。左侧上颌组织进行组织学和免疫组织化学分析,检测牙槽嵴顶附近RANKL及OPG的表达。用抗酒石酸酸性磷酸酶(Tartrate-resistant acid phosphatasem,TRAP)检测OC的数目。结果P组与N组相比,ABL增加、结合上皮(junctional epithelium,JE)根方迁移、牙槽骨吸收明显,表明P组牙周炎建模成功。E组与P组相比,ABL减少,炎症细胞浸润减少,RANKL免疫标记细胞及TRAP阳性多核细胞减少,OPG基因表达上调(P0.05)。而E组与N组相比,ABL和OC数目差异无统计学意义(P0.05)。结论β-隐黄素影响牙周炎的发生发展,抑制了牙周炎牙槽骨丧失。其可能的作用机制是通过下调RANKL/OPG的比值,减少OC的数目而发挥作用。
[Abstract]:Background in the process of periodontitis, the presence of inflammatory cytokines and chemical factors up-regulates the expression of nuclear factor 魏 B receptor activating factor ligand Receptor activator of nuclear factor-kappa B ligand RANKL and RANKL activated osteoclast (Osteoclastus) precursor cells. Then the matured polynuclear OC adsorbed on the surface of the mineralized bone matrix for bone resorption. Alveolar bone resorption is the main cause of tooth loosening and abscission. Preventing alveolar bone resorption and promoting alveolar bone repair and regeneration are the key to cure periodontitis. 尾 -Cryptoflavin, a carotenoid found in fruits and vegetables, can form bone by stimulating osteoblasts. Inhibition of OC absorption of bone to prevent loss of bone tissue. Previous studies have found that the low level of serum 尾 -cryptoflavin is associated with an increase in the prevalence of periodontitis, and that the serum level of 尾 -cryptoflavin is significantly lower in patients with periodontitis, and that 尾 -cryptoflavin can inhibit the inflammatory response induced by endotoxin lipopolysaccharide (LPSs). The ratio of osteoprotegerin / RANKL was up-regulated, and 尾 -cryptoflavin could inhibit the decrease of bone mineral density induced by LPS-induced bone density. These studies suggest that 尾-Cryptoflavin affects the occurrence and development of periodontitis. However, the effect of 尾 -Cryptoflavin on alveolar bone resorption in periodontitis is less studied, and its mechanism is not very clear. Objective to evaluate the effect of 尾 -cryptoflavin on periodontal tissue in rats with experimental periodontitis by detecting the number of alveolar bone loss and the expression of RANKL and OPG in alveolar bone. To detect the effect on alveolar bone resorption in periodontitis and analyze its mechanism. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: control group (n = 1) and control group (n = 2). Induction of experimental periodontitis model: orthodontic ligation of diameter 0.2mm was performed around the neck of bilateral maxillary second molar, and Escherichia coli LPS(30 渭 l / rat was injected into the gingival sulcus of buccal side, once every 48 hours, for 3 times. 尾 -Cryptoflavin injected 12 渭 l of 尾 -Cryptoflavin at the same site after injection of LPS, once every 48 hours, and the same amount of corn oil was injected into the group of 尾 -Cryptoflavin at the same site after the intervention of 尾 -Cryptoflavin once every 48 hours. Rats were killed with 10% chloral hydrate injected intraperitoneally on the 8th day of the experiment, and bilateral maxillary tissues (including teeth, gingiva, periodontal membrane and alveolar bone) were taken. The distance between Cemento-enamel junction (CEJ) and Alveolar bone crestus (ABC) was measured. The expression of RANKL and OPG near alveolar crest was detected by histological and immunohistochemical analysis in left maxillary tissue. The number of OC was detected by tartrate-resistant acid phosphatasemem TRAPs resistant to tartaric acid phosphatase. Results compared with group N, the ABL in group P was increased, the root side migration and alveolar bone resorption were obvious, which indicated that the ABL of periodontitis in group P was less than that in group P. Inflammatory cell infiltration decreased RANKL immunolabeled cells and TRAP positive multinucleated cells decreased the expression of OPG gene upregulated (P0.05). There was no significant difference in the number of ABL and OC between group E and group N (P 0.05). Conclusion 尾 -cryptoflavin affects the occurrence and development of periodontitis and inhibits the loss of alveolar bone in periodontitis. Its possible mechanism is to reduce the number of OC by down-regulating the ratio of RANKL/OPG.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781.42

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