APN-1在糖尿病牙周炎小鼠牙龈组织的表达
发布时间：2018-10-22 08:11
【摘要】：研究发现,低脂联素血症与T2DM发展过程中的高胰岛素血症和胰岛素抵抗有关。故有学者提出,脂联素也可作为探讨患者将来发生糖尿病的一个独立因子。在牙周炎和糖尿病的发生发展机制过程中,炎症反应失调和加剧是关键,有研究报道,对伴有糖尿病牙周炎患者进行牙周干预治疗后,患者血清脂联素水平上升,牙周及全身状况得到改善,有可能减少糖尿病及其并发症的发生和发展。然而,糖尿病伴牙周炎患者的局部炎症状况及其参与机体胰岛素抵抗的发生发展是否均与脂联素水平变化有关,这些问题都尚未得到解决。通过动物实验,建立伴有糖尿病的牙周炎动物模型,研究脂联素与血糖水平及炎症状态是否具有相关性,为研究糖尿病与牙周炎的相关性及日益增多的糖尿病伴有牙周炎患者的临床治疗及预防提供一些理论依据。 目的：通过检测健康、糖尿病及糖尿病伴牙周炎昆明系小鼠牙龈组织中脂联素基因及蛋白含量的变化,检测三组鼠的血浆中脂联素含量,探讨脂联素含量与血糖水平及牙周炎症状态是否具有相关性。 方法：45只3周龄的昆明系小鼠按不同造模方式分为三组： A组对照组(15只),标准颗粒饲料喂养。 B组糖尿病组(15只)。 15只健康鼠通过四氧嘧啶(Alloxan)注射法4周建立小鼠糖尿病模型(12mg/100g体重来诱导)。 C组糖尿病+牙周炎模型(15只)。 15只健康鼠先与B组同时建糖尿病模型,糖尿病组造模成功后左、右上颌第一磨牙及第二磨牙牙颈部丝线结扎联合高糖喂养6周建立小鼠糖尿病+牙周炎模型。 通过测检AB两组小鼠不同时间的血糖、空腹胰岛素水平(Fins)及胰岛素抵抗指数,并与空白对照组对比来确认小鼠糖尿病组造模成功。通过牙周组织病理组织学检查及CBCT扫描观察牙槽骨丧失程度判断小鼠糖尿病+牙周炎模型造模成功。 分别取三组小鼠的牙龈组织,用RT-PCR、western blot检测牙龈组织中脂联素基因及蛋白含量的变化,并于鼠处死时取三组鼠的血浆,通过ELISA方法测量血浆中脂联素含量进行对比和统计学分析。 结果： 1.本实验发现,B组造模后20天,小鼠血糖、空腹胰岛素水平(Fins)及胰岛素抵抗指数大于正常值,与空白对照组相比差异具有统计学意义(P0.01),小鼠糖尿病组造模成功。 2.牙龈组织病理组织学检测显示糖尿病伴牙周炎组小鼠牙龈炎症表现最明显、在单纯糖尿病组部分小鼠可见牙龈组织轻度炎症、健康组小鼠牙龈未见明显炎症；CBCT扫描结果显示糖尿病伴牙周炎组小鼠牙槽骨丧失程度最明显,糖尿病组小鼠次之,健康组小鼠未见明显骨丧失。小鼠糖尿病+牙周炎组造模成功。 3.RT-PCR、western blot检测小鼠牙龈组织中脂联素基因及蛋白表达水平在三组间比较均有差异,存在统计学意义(P0.05)。在健康组表达水平最强,在糖尿病组、糖尿病伴牙周炎组小鼠中表达水平依次减弱。 4. ELISA结果显示APN蛋白在健康组小鼠血浆中表达水平最强,在糖尿病组、糖尿病伴牙周炎组小鼠血浆中表达水平依次减弱;组间比较均有差异,存在统计学意义(P0.05)。 结论： 1.可通过四氧嘧啶(Alloxan)注射法有效建立小鼠糖尿病模型。可通过牙周局部结扎联合高糖喂养有效建立小鼠糖尿病伴牙周炎模型。 2.糖尿病伴牙周炎组小鼠与糖尿病组小鼠、健康组小鼠相比,牙龈炎症及骨丧失程度最明显,脂联素基因和蛋白含量在小鼠牙龈组织中表达显著降低。提示糖尿病可能通过降低脂联素水平促进牙龈局部炎症的发生、发展；与此同时,牙周炎可能协同降低脂联素水平,促进糖尿病的发展。 3.糖尿病后牙龈组织及血浆中脂联素表达降低可能是牙周炎发病及加重的重要因素之一,APN可能在糖尿病和牙周炎相互影响关系中起着一定作用,是糖尿病的保护性因子,且有一定的抗炎作用。为临床治疗和预防牙周炎提供理论依据。
[Abstract]:It has been found that hypolipoproteinemia is associated with hyperinsulinemia and insulin resistance in the progression of T2DM. Therefore, it is suggested that adiponectin can also be used as an independent factor to explore the future occurrence of diabetes in patients. In the pathogenesis of periodontitis and diabetes, inflammatory response and intensification are key, and studies have been conducted to improve serum adiponectin levels, periodontal and systemic conditions after periodontal intervention in patients with diabetic periodontitis. It is possible to reduce the occurrence and development of diabetes and its complications. However, the local inflammatory conditions of diabetic patients with periodontitis and their involvement in the development of body insulin resistance are related to changes in adiponectin levels, which have not been resolved. animal model of periodontitis with diabetes is established by animal experiment to study whether adiponectin is relevant to blood sugar level and inflammation state, To study the correlation between diabetes mellitus and periodontitis and the increasing number of diabetes patients with periodontitis, some theoretical bases are provided. Objective: To investigate the changes of adiponectin gene and protein content in gingival tissues of Kunming mice with periodontitis and periodontitis. Methods: Forty-five-week-old Kunming mice were treated with different models. The formula is divided into three groups: A group control group (15 only), Standard pellet feed. Group B diabetes mellitus group (15 rats). 15 healthy rats established the model of diabetes mellitus (1) in 4 weeks by Alloxin injection method. 2mg/ 100g body weight). Group C diabetes + periodontitis model (15 rats). 15 healthy rats were first established with group B while the model of diabetes mellitus was constructed. The model of diabetic + periodontitis in mice was established by high sugar feeding for 6 weeks. The blood glucose, fasting insulin level (Fins) and insulin resistance index of two groups of mice were measured. and compared with the blank control group to confirm the success of the model of the diabetic group of the mice. The pathological histology examination of the periodontal tissue and the CBCT scanning observation teeth To determine the degree of loss of bone loss, the model of diabetic + periodontitis was established successfully. The gingival tissues of three groups of mice were taken respectively, and the changes of adiponectin gene and protein content in gingival tissues were detected by RT-PCR and Western blot. LIS The content of adiponectin in plasma was compared and analyzed by A method. Results: 1. The results showed that the blood glucose, fasting insulin level (Fins) and insulin resistance index of mice were more than normal in 20 days after group B modeling and blank Compared with the control group, the difference was statistically significant (P0.01), and the model of diabetic group was successful. There was no significant inflammation in the gum tissue in the healthy group of mice in the pure diabetic group, and the CBCT scan showed diabetes mellitus with periodontitis. The alveolar bone loss of the group was the most obvious, the sugar The mice showed no obvious bone loss in the group of urine disease group. The model of diabetic + periodontitis group was successful. RT-PCR and western blot were used to detect the gingival tissues of mice. There was a significant difference in the levels of adiponectin gene and protein expression among the three groups (P0.05). in diabetic group, that expression level of APN protein in healthy group of mice was the strongest, and the expression level of APN protein in healthy group was the strongest in diabetic group and diabetic group. periodontitis The levels of expression in plasma of group mice were decreased in turn; there was a difference between the groups, and there was statistical significance (P 0. 05). Conclusion: 1. Available through Alloxa n) The mice model of diabetes mellitus can be effectively established by injection method. The model of diabetic companion periodontitis can be effectively established by the combination of periodontal local ligation and high-sugar feeding. Compared with the group of mice and healthy mice, the degree of gingival inflammation and bone loss was most obvious, and the expression of adiponectin gene and protein in the gingival tissues of mice was significantly decreased. Decrease the level of adiponectin to promote local inflammation of the gum, develop; At the same time, periodontitis may work synergistically to reduce the level of adiponectin and promote the development of diabetes. One of the factors is that APN may be in diabetes and periodontal
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R781.42
本文编号：2286607
[Abstract]:It has been found that hypolipoproteinemia is associated with hyperinsulinemia and insulin resistance in the progression of T2DM. Therefore, it is suggested that adiponectin can also be used as an independent factor to explore the future occurrence of diabetes in patients. In the pathogenesis of periodontitis and diabetes, inflammatory response and intensification are key, and studies have been conducted to improve serum adiponectin levels, periodontal and systemic conditions after periodontal intervention in patients with diabetic periodontitis. It is possible to reduce the occurrence and development of diabetes and its complications. However, the local inflammatory conditions of diabetic patients with periodontitis and their involvement in the development of body insulin resistance are related to changes in adiponectin levels, which have not been resolved. animal model of periodontitis with diabetes is established by animal experiment to study whether adiponectin is relevant to blood sugar level and inflammation state, To study the correlation between diabetes mellitus and periodontitis and the increasing number of diabetes patients with periodontitis, some theoretical bases are provided. Objective: To investigate the changes of adiponectin gene and protein content in gingival tissues of Kunming mice with periodontitis and periodontitis. Methods: Forty-five-week-old Kunming mice were treated with different models. The formula is divided into three groups: A group control group (15 only), Standard pellet feed. Group B diabetes mellitus group (15 rats). 15 healthy rats established the model of diabetes mellitus (1) in 4 weeks by Alloxin injection method. 2mg/ 100g body weight). Group C diabetes + periodontitis model (15 rats). 15 healthy rats were first established with group B while the model of diabetes mellitus was constructed. The model of diabetic + periodontitis in mice was established by high sugar feeding for 6 weeks. The blood glucose, fasting insulin level (Fins) and insulin resistance index of two groups of mice were measured. and compared with the blank control group to confirm the success of the model of the diabetic group of the mice. The pathological histology examination of the periodontal tissue and the CBCT scanning observation teeth To determine the degree of loss of bone loss, the model of diabetic + periodontitis was established successfully. The gingival tissues of three groups of mice were taken respectively, and the changes of adiponectin gene and protein content in gingival tissues were detected by RT-PCR and Western blot. LIS The content of adiponectin in plasma was compared and analyzed by A method. Results: 1. The results showed that the blood glucose, fasting insulin level (Fins) and insulin resistance index of mice were more than normal in 20 days after group B modeling and blank Compared with the control group, the difference was statistically significant (P0.01), and the model of diabetic group was successful. There was no significant inflammation in the gum tissue in the healthy group of mice in the pure diabetic group, and the CBCT scan showed diabetes mellitus with periodontitis. The alveolar bone loss of the group was the most obvious, the sugar The mice showed no obvious bone loss in the group of urine disease group. The model of diabetic + periodontitis group was successful. RT-PCR and western blot were used to detect the gingival tissues of mice. There was a significant difference in the levels of adiponectin gene and protein expression among the three groups (P0.05). in diabetic group, that expression level of APN protein in healthy group of mice was the strongest, and the expression level of APN protein in healthy group was the strongest in diabetic group and diabetic group. periodontitis The levels of expression in plasma of group mice were decreased in turn; there was a difference between the groups, and there was statistical significance (P 0. 05). Conclusion: 1. Available through Alloxa n) The mice model of diabetes mellitus can be effectively established by injection method. The model of diabetic companion periodontitis can be effectively established by the combination of periodontal local ligation and high-sugar feeding. Compared with the group of mice and healthy mice, the degree of gingival inflammation and bone loss was most obvious, and the expression of adiponectin gene and protein in the gingival tissues of mice was significantly decreased. Decrease the level of adiponectin to promote local inflammation of the gum, develop; At the same time, periodontitis may work synergistically to reduce the level of adiponectin and promote the development of diabetes. One of the factors is that APN may be in diabetes and periodontal
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R781.42
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相关期刊论文 前2条
1 沙月琴 ,耿素芳 ,陈智滨 ,张京华 ,李葳;牙周基础治疗对糖化血红蛋白及胶原酶的影响[J];现代口腔医学杂志;2003年01期
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