壳寡糖对骨质疏松伴牙周炎大鼠骨代谢及IKK/NF-κB通路的影响

doi:10.19439/j.sjos.2023.02.005

摘要/Abstract

摘要： 目的: 探讨壳寡糖对骨质疏松伴牙周炎模型小鼠骨代谢和IKK/NF-κB通路的影响。方法: 将30只大鼠随机分为3组,每组10只,分为对照组、去势牙周炎组和壳寡糖治疗组。除对照组外,其余2组大鼠均切除卵巢并涂抹牙龈卟啉单胞菌液,建立骨质疏松伴牙周炎模型。结扎手术后4周,以200 mg/kg灌胃壳寡糖治疗组大鼠,另2组灌胃等体积生理盐水,每天1次,连续90 天。观察各组牙周组织变化,给药前、给药结束后检测大鼠骨密度。给药结束后,尾静脉采血,采用酶联免疫吸附法测定各组血清碱性磷酸酶（ALP）、骨钙素（BGP）和抗酒石酸酸性磷酸酶5b（TRACP5b）的含量。采用视诊和探诊相结合的方法得到各组大鼠的牙龈指数和牙周附着丧失;处死大鼠,取出上颌骨,测量釉-牙骨质界到牙槽嵴顶的距离,得到牙槽骨吸收值。采用H-E染色,对各组大鼠上颌骨牙体、牙周组织进行病理学观察。采用RT-PCR和蛋白免疫印迹法分别检测各组大鼠牙周组织中核因子κB（NF-κB）、IκB激酶（IKK）mRNA和蛋白表达水平。采用SPSS 22.0软件包对数据进行统计学分析。结果: 对照组大鼠牙龈呈粉红色,探诊不出血;去势牙周炎组大鼠牙龈红肿,轻触出血;壳寡糖治疗组大鼠牙龈轻微泛红,探诊不出血。给药结束后,与对照组相比,去势牙周炎组大鼠的骨密度、血清ALP和BGP显著降低（P<0.05）,TRACP5b、牙龈指数、牙周附着丧失和牙槽骨吸收、牙周组织NF-κB和IKK mRNA和蛋白表达水平显著上升（P<0.05）。与去势牙周炎组相比,壳寡糖治疗组大鼠的骨密度、血清ALP和BGP显著升高（P<0.05）,TRACP5b、牙龈指数、牙周附着丧失和牙槽骨吸收、牙周组织NF-κB和IKKmRNA和蛋白表达水平显著下降（P<0.05）。去势牙周炎组大鼠牙周组织结合上皮与牙面分离,牙周袋明显且深,牙槽骨高度降低;壳寡糖治疗组大鼠牙周组织虽可观察到牙周袋,但不明显,并且牙槽骨周围出现新骨。结论: 壳寡糖可诱导骨代谢生化指标趋于正常,缓解牙周炎症状,可能与壳寡糖抑制IKK/NF-κB通路有关。

关键词: 骨质疏松, 牙周炎, 壳寡糖, 骨代谢, 牙槽骨吸收

Abstract: PURPOSE: To investigate the effects of chitosan oligosaccharide on bone metabolism and IKK/NF-κB pathway in mice with osteoporosis and periodontitis. METHODS: Thirty rats were randomly divided into 3 groups, with 10 rats in each group. They were divided into control group, ovariectomized periodontitis group and chitosan oligosaccharide treatment group. Except for the control group, the other two groups were ovariectomized and smeared with Porphyromonas gingivalis fluid to establish the model of osteoporosis with periodontitis. Four weeks after ligation, the rats in chitosan oligosaccharide treatment group were gavaged with 200 mg/kg chitosan oligosaccharide, and the other two groups were gavaged with equal volume of normal saline once a day for 90 days. The periodontal tissues of each group were observed before administration, and the bone mineral density of rats was detected by dual energy X-ray animal bone mineral density and body composition analysis system. After 90 days of administration, the bone mineral density was detected again. After administration, blood was collected from tail vein, and the contents of serum alkaline phosphatase (ALP), bone Gla protein (BGP) and tartrate resistant acid phosphatase 5b (TRACP5b) were measured by enzyme-linked immunodeficient assay. The gingival index and periodontal attachment loss of rats in each group were obtained by visual examination and exploratory examination. The maxilla was removed, and the distance from the enamel cementum boundary to the alveolar crest was measured to obtain alveolar bone absorption value. H-E staining was used to observe the pathology of maxilla in each group. RT-PCR and Western blot were used to detect the nuclear factors in periodontal tissue of rats in each group. SPSS 22.0 software package was used for statistical analysis. RESULTS: Before administration, the gums of the control group were pink without bleeding, and the gums of the other two groups were red and swollen with slight bleeding. After administration, compared with the control group, the bone mineral density, serum ALP, BGP of ovariectomized periodontitis group decreased significantly(P<0.05); while TRACP5b, gingival index, loss of periodontal attachment and alveolar bone resorption, NF-κB and IKK mRNA and protein expression in periodontal tissue increased significantly(P<0.05). Compared with the ovariectomized periodontitis group, the bone mineral density, serum ALP, BGP were significantly increased(P<0.05); while TRACP5b, gingival index, periodontal attachment loss and alveolar bone resorption, NF-κB and IKK mRNA and protein expression in periodontal tissue were significantly decreased (P<0.05). In the ovariectomized periodontitis group, the periodontal tissue combined with epithelium was separated from the tooth surface, the dental pocket was obvious and deep, and the height of alveolar bone decreased. Although dental pocket could be observed in the periodontal tissue of rats treated with chitosan oligosaccharide, it was not obvious, and new bone appeared around the alveolar bone. CONCLUSIONS: Chitosan oligosaccharide can induce biochemical indexes of bone metabolism to become normal, alleviate the symptoms of periodontitis, this may be related to the inhibition of IKK/NF-κB pathway by chitosan oligosaccharide.

Key words: Osteoporosis, Periodontitis, Chitosan oligosaccharide, Bone metabolism, Alveolar bone resorption

中图分类号:

R781.4

俞晓霏, 汪信洋, 李斌, 陈耀忠. 壳寡糖对骨质疏松伴牙周炎大鼠骨代谢及IKK/NF-κB通路的影响[J]. 上海口腔医学, 2023, 32(2): 137-142.

YU Xiao-fei, WANG Xin-yang, LI Bin, CHEN Yao-zhong. Effects of chitosan oligosaccharide on bone metabolism and IKK/NF-κB pathway in rats with osteoporosis and periodontitis[J]. Shanghai Journal of Stomatology, 2023, 32(2): 137-142.

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