片仔癀对糖尿病牙周炎大鼠Keap1/Nrf2通路及牙槽骨丢失的影响

Effects of PienTzeHuang on Keap1/Nrf2 pathway and alveolar bone loss in diabetic periodontitis rats

目的:探讨片仔癀对糖尿病牙周炎大鼠Kelch样ECH相关蛋白1(Keap1)/核因子红血球相关因子2(Nrf2)通路及牙槽骨丢失的影响。方法:建立糖尿病牙周炎大鼠模型,随机分为模型组、牙周基础治疗组、片仔癀低剂量(0.75 g/kg)+牙周基础治疗组、片仔癀中剂量(1.5 g/kg)+牙周基础治疗组、片仔癀高剂量(3.0 g/kg)+牙周基础治疗组,每组12只,另取12只设为对照组。分组治疗后,以牙龈指数评估各组大鼠牙周组织炎症情况;通过断层扫描检测各组大鼠牙槽骨吸收值;通过HE染色检测各组大鼠牙周组织病理变化;通过试剂盒检测各组大鼠牙周组织超氧化物歧化酶(SOD)和丙二醛(MDA)水平;通过ELISA法检测各组大鼠血清中IL-6和TNF-α水平;通过Western blot检测各组大鼠牙周组织Keap1/Nrf2通路相关蛋白(Keap1、Nrf2)表达水平。结果:与对照组相比,模型组大鼠牙周组织牙周膜结构紊乱,纤维束排列异常,出现断裂,且伴随炎症细胞大量浸润等病理损伤,牙龈指数、牙槽骨吸收值、MDA、IL-6、TNF-α、Keap1蛋白表达水平明显升高,同时SOD水平、Nrf2蛋白表达水平明显降低,差异有统计学意义(P<0.05);与模型组相比,牙周基础治疗组、片仔癀低剂量+牙周基础治疗组、片仔癀中剂量+牙周基础治疗组、片仔癀高剂量+牙周基础治疗组大鼠牙周组织病理损伤均减轻,牙龈指数、牙槽骨吸收值、MDA、IL-6、TNF-α、Keap1蛋白表达水平均降低,同时SOD水平、Nrf2蛋白表达水平均升高,差异有统计学意义(P<0.05);与牙周基础治疗组相比,片仔癀低剂量+牙周基础治疗组、片仔癀中剂量+牙周基础治疗组、片仔癀高剂量+牙周基础治疗组大鼠牙周组织病理损伤均减轻,牙龈指数、牙槽骨吸收值、MDA、IL-6、TNF-α水平、Keap1蛋白表达水平均降低,同时SOD水平、Nrf2蛋白表达水平均升高,且片仔癀各组呈剂量依赖性,差异有统计学意义(P<0.05)。结论:片仔癀可下调Keap1蛋白表达,上调Nrf2蛋白表达,抑制糖尿病牙周炎大鼠牙周氧化应激及炎症反应,改善其牙槽骨丢失症状。

Objective:To investigate the effect of PienTzeHuang on Keap1/Nrf2 pathway and alveolar bone loss in rats with diabetic periodontitis.Methods:Diabetic periodontitis rat model was established and randomly divided into model group,periodontal basic treatment group,low-dose(0.75 g/kg)PienTzeHuang+periodontal basic treatment group,medium-dose(1.5 g/kg)PienTzeHuang+periodontal basic treatment group,high-dose(3.0 g/kg)PienTzeHuang+periodontal basic treatment group,with 12 rats in each group,another 12 rats were set as control group.After treatment,the gingival index was used to evaluate periodontal inflammation;the alveolar bone resorption of rats in each group were detected by tomography;the pathological changes of periodontal tissue were detected by HE staining;the levels of SOD and MDA in periodontal tissue were detected by kit;the serum levels of IL-6 and TNF-αwere detected by ELISA;the expression levels of Keap1/Nrf2 pathway related proteins(Keap1,Nrf2)in periodontal tissues were detected by Western blot.Results:Compared with control group,the periodontal ligament structure of the model group was disordered,the fiber bundles were arranged abnormally and fractured,accompanied by a large number of inflammatory cells infiltration and other pathological damage,the gingival index,alveolar bone resorption,MDA level,IL-6 level,TNF-αlevel and Keap1 protein expression level were significantly higher,at the same time,SOD level and Nrf2 protein expression level were significantly lower(P<0.05);compared with model group,the pathological damage of periodontal tissue was reduced in the periodontal basic treatment group,the low-dose PienTzeHuang+basic periodontal treatment group,the medium-dose PienTzeHuang+basic periodontal treatment group,and the high-dose PienTzeHuang+basic periodontal treatment group,the gingival index,alveolar bone resorption,MDA,IL-6,TNF-αlevel and Keap1 protein expression level were significantly lower,at the same time,SOD level and Nrf2 protein expression level were significantly higher(P<0.05);compared with periodontal basic treatment group,the pathological damage of periodontal tissue was reduced in the low-dose PienTzeHuang+basic periodontal treatment group, the medium-dose PienTzeHuang+basic periodontaltreatment group, and the high-dose PienTzeHuang+basic periodontal treatment group, the gingival index, alveolar bone resorption,MDA level, IL-6 level, TNF- α level and Keap1 protein expression level were significantly lower, at the same time, SOD level andNrf2 protein expression level were significantly higher, and the PienTzeHuang groups were dose-dependent( P<0.05). Conclusion:PienTzeHuangcan down-regulate the expression of Keap1 protein and up-regulate the expression of Nrf2 protein, inhibit periodontal oxidativestress and inflammatory reaction, and improve the symptoms of alveolar bone loss in diabetic periodontitis rats.