盐酸小檗碱调节大鼠实验性牙周炎牙龈组织MMPs/TIMP表达变化及其机制
Berberine regulates MMPs/TIMP via inhibition of P38 MAPK/NF-κB phosphorylation and anti-inflammation in rat experimental periodontitis

摘要：目的 探讨盐酸小檗碱对牙周炎基质金属蛋白酶/基质金属蛋白酶组织抑制物(MMPs/TIMP)的调控及其机制。方法 采用丝线结扎+菌液注射法建立大鼠牙周炎模型后，给予150mg/(kg？d)盐酸小檗碱或生理盐水干预。显微X射线断层计算机扫描(μ-CT)和苏木素-伊红(HE)染色观察牙周组织学改变，Western blot检测大鼠牙龈组织MMP-2、MMP-9、TIMP-2的蛋白表达及P38 MAPK/NF-κB磷酸化水平，ELISA测定牙龈组织肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)、白介素-1β(interleukin-1β, IL-1β)的含量。结果 牙周检查发现牙周炎组大鼠牙龈红肿明显，触之易出血，牙周袋较深，μ-CT显示牙槽骨吸收明显，牙龈组织TNF-α和IL-1β含量显著增加，MMP-2和MMP-9表达增多，并伴有磷酸化P38 MAPK/NF-κB上调。盐酸小檗碱治疗后可明显改善牙周情况，减少牙槽骨吸收，降低牙龈组织TNF-α和IL-1β含量，抑制P38 MAPK/NF-κB磷酸化，最终降低MMP-2和MMP-9的过度表达并提高TIMP-2的表达。结论 盐酸小檗碱可能通过抑制P38 MAPK/NF-κB磷酸化，降低牙龈组织TNF-α和IL-1β，调节牙龈组织MMPs/TIMP的过量表达而发挥对大鼠牙周炎的治疗作用。
ABSTRACT: Objective To investigate the effects of berberine on matrix metalloproteinases/tissue inhibitors of matrix metalloproteinases (MMPs/TIMPs) in rat periodontitis. Methods Adult male Sprague-Dawley rats were subjected to thread ligation and porphyromonas gingivalis (P. gingivalis) inoculation resulting in periodontitis. Berberine \[150mg/(kg？d)\] or vehicle was administered by oral gavage for 4 weeks. Alveolar bone loss and soft-tissue destruction were determined by micro-computed tomography (μ-CT) and HE staining. The contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) of gingival tissue were examined by ELISA. The expressions of MMP-2, MMP-9 and TIMP-2 as well as the phosphorylation of P38 MAPK and NF-κB were determined by Western blot. Results Significant increases were observed in alveolar bone loss and periodontal soft-tissue destruction with higher levels of TNF-α, IL-1β and MMP-2/9 expressions and the activities of P38 MAPK and NF-κB in the experiment periodontitis group. Berberine of \[150mg/(kg？d)\] not only improved the periodontal tissue and decreased alveolar bone loss, but also reduced the contents of TNF-α, IL-1β and MMP-2/9 and increased TIMP-2. In addition, berberine inhibited the phosphorylation of P38 MAPK/NF-κB. Conclusion Berberine protects against periodontal tissue damage via decreasing MMP-2 and MMP-9 expressions but increasing TIMP-2 expression, which may be induced by inhibition of P38 MAPK/NF-κB and the anti-inflammatory effect on rat periodontitis