吉林大学学报(医学版) ›› 2023, Vol. 49 ›› Issue (1): 31-38.doi: 10.13481/j.1671-587X.20230105

• 基础研究 • 上一篇    下一篇

葡萄籽原花青素提取物对糖尿病牙周炎大鼠牙周组织炎症的缓解作用及其对牙周组织中TLR4和NF-κB表达水平的影响

朱晓娟,代海涛,李艳,崔灵欣,王亚,徐江,仵楠()   

  1. 石河子大学医学院第一附属医院口腔科,新疆 石河子 832000
  • 收稿日期:2022-02-20 出版日期:2023-01-28 发布日期:2023-02-03
  • 通讯作者: 仵楠 E-mail:195773220@qq.com
  • 作者简介:朱晓娟(1995-),女,安徽省亳州市人,住院医师,医学硕士,主要从事牙周病防治方面的研究。
  • 基金资助:
    国家自然科学基金项目(82160572);石河子大学科研基金项目(ZZZC201819A)

Improvement effect of grape seed proanthocyanidin extract on periodontal inflammation in diabetic periodontitis rats and its influence on expression levels of TLR4 and NF-κB in periodontal tissue

Xiaojuan ZHU,Haitao DAI,Yan LI,Lingxin CUI,Ya WANG,Jiang XU,Nan WU()   

  1. Department of Stomatology,First Affiliated Hospital,Shihezi Medical University,Shihezi 832000,China
  • Received:2022-02-20 Online:2023-01-28 Published:2023-02-03
  • Contact: Nan WU E-mail:195773220@qq.com

摘要:

目的 探讨葡萄籽原花青素提取物(GSPE)对糖尿病牙周炎大鼠牙周炎症的改善作用,并阐明其作用机制。 方法 40只雄性SD大鼠随机分为对照组、糖尿病牙周炎模型组(模型组)、低剂量(100 mg·kg-1)GSPE组和高剂量(200 mg·kg-1)GSPE组,每组10只,除对照组外,其他各组大鼠采用牙周结扎法联合一次性腹腔注射链脲佐菌素(STZ)建立糖尿病牙周炎模型。模型建立成功后,低和高剂量GSPE组大鼠分别按100和200 mg·kg-1GSPE灌胃给药,对照组和模型组大鼠给予等量生理盐水灌胃,每天1次,连续8周。观察大鼠一般情况和血糖水平变化,采用HE染色观察各组大鼠牙周组织病理形态表现,测量各组大鼠牙槽骨吸收量,采用相关试剂盒检测各组大鼠牙周组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和活性氧(ROS)及丙二醛(MDA)水平,酶联免疫吸附试验(ELISA)法检测各组大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)水平, Western blotting法检测各组大鼠牙周组织中Toll样受体4(TLR4)和核因子κB(NF-κB)蛋白表达水平。 结果 与对照组比较,模型组大鼠出现明显“三多一少”症状,血糖水平明显升高(P<0.05),牙周结缔组织中炎性细胞浸润,牙周组织损伤,牙槽骨吸收量明显增加(P<0.05),牙周组织中SOD和CAT活性明显降低(P<0.05),ROS和MDA水平明显升高(P<0.05),血清中TNF-α、IL-1β和IL-6水平明显升高(P<0.05),牙周组织中TLR4和NF-κB蛋白表达水平明显升高(P<0.05);与模型组比较,低和高剂量GSPE组大鼠一般情况均明显好转,血糖水平明显降低(P<0.05),牙周组织损伤明显减轻,牙槽骨吸收量减少(P<0.05),牙周组织中SOD和CAT活性明显升高(P<0.05),ROS和MDA水平明显降低(P<0.05),血清中TNF-α、IL-1β和IL-6水平明显降低(P<0.05),牙周组织中TLR4和NF-κB 蛋白表达水平明显降低(P<0.05);与低剂量GSPE组比较,高剂量GSPE组大鼠牙周组织中SOD和CAT活性明显升高(P<0.05),ROS和MDA水平明显降低(P<0.05),血清中TNF-α、IL-1β和IL-6水平明显降低(P<0.05),牙周组织中TLR4和NF-κB 蛋白表达水平明显降低(P<0.05)。 结论 GSPE可改善糖尿病牙周炎大鼠牙周组织炎症,其作用机制与调节牙周组织中TLR4/NF-κB信号通路有关。

关键词: 葡萄籽原花青素, 糖尿病, 牙周炎, Toll样受体4, 核因子κB

Abstract:

Methods Forty male SD rats were randomly divided into control group, diabetic periodontitis model group(model group), low dose (100 mg·kg-1) of GSPE group and high dose (200 mg·kg-1) of GSPE group; there were 10 rats in each group. Except for control group, the rats in other groups were used to establish the diabetic periodontitis models by periodontal ligation combined with intraperitoneal injection of streptozotocin (STZ). After the successful establishment of the models, the rats in low and high doses of GSPE groups were given 100 and 200 mg·kg-1 GSPE, respectively, while the rats in control group and model group were given the same amount of normal saline once a day for 8 weeks. The general situation of the rats and the changes of blood glucose levels were observed, the pathomorphology of periodontal tissue of the rats in various groups were observed by HE staining, the alveolar bone absorption of the rats in various groups was measured, the activities of superoxide dismutase (SOD) and the catalase (CAT) and the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in periodontal tissue of the rats in various groups were detected by related kits.Enzyme-linked immunosorbent assay(ELISA) method was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) in serum of the rats in various groups. The expression levels of Toll-like receptor 4(TLR4) and nuclear factor-κB(NF-κB) proteins in periodontal tissue of the rats in various groups were detected by Western blotting method. Results Compared with control group, the rats in model group showed obvious symptoms of “three more and one less”;the level of blood glucose of the rats was increased obviously(P<0.05), the inflammatory cells were infiltrated in periodontal connective tissue, periodontal tissue was damaged, and the alveolar bone absorption amount was increased(P<0.05);the activities of SOD and CAT in periodontal tissue were significantly decreased (P<0.05), the levels of ROS and MDA were significantly increased (P<0.05), the levels of TNF-α, IL-1β and IL-6 in serum were significantly increased(P<0.05),and the expression levels of TLR4 and NF-κB proteins were significantly increased (P<0.05). Compared with model group, the general condition of the rats in low and high doses of GSPE groups was obviously improved,the levels of blood glucose were reduced(P<0.05), periodontal tissue damage was obviously reduced,the alveolar bone absorption amounts were reduced(P<0.05), the activities of SOD and CAT in periodontal tissue were significantly increased (P<0.05), the levels of ROS and MDA were significantly decreased (P<0.05), the levels of TNF-α, IL-1β and IL-6 in serum were significantly decreased(P<0.05),and the expression levels of TLR4 and NF-κB proteins were significantly decreased (P<0.05). Compared with low dose of GSPE group, the activities of SOD and CAT in periodontal tissue of the rats in high dose of GSPE group were significantly increased (P<0.05), the levels of ROS and MDA were significantly decreased (P<0.05), the levels of TNF-α, IL-1β and IL-6 in serum were significantly decreased(P<0.05),and the expression levels of TLR4 and NF-κB proteins were significantly decreased (P<0.05). Conclusion GSPE can improve the periodontal inflammation in the diabetic periodontitis rats, and its mechanism may be related to regulating the TLR4 and NF-κB signaling pathway in periodontal tissue. Objective To explore the improvement effects of grape seed proanthocyanidin extract (GSPE) on the periodontal inflammation in the diabetic periodontitis rats, and to clarify its mechanism.

Key words: Grape seed proanthocyanidin extract, Diabetes mellitus, Periodontitis, Toll-like receptor 4, Nuclear factor-κB

中图分类号: 

  • R781.4