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

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。

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 [150 mg/（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 [150 mg/（kg · d）] not only improved the periodontal tissue and decreased alveolar bone loss ,but also reduced the contents of TNF‐α,IL‐1βand 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 .