小檗碱对糖尿病大鼠牙周炎影响的实验观察

Influence of Berberine on Periodontitis in Diabetic Rats

目的:验证小檗碱是否有助于治疗大鼠糖尿病伴发牙周炎。方法:30只雄性SD大鼠随机等分为3组,即对照组(C组)、小檗碱组(B组)和糖尿病组(D组)。B组和D组采用高糖饮食+链脲佐菌素构建大鼠糖尿病模型;建模后第3天,B组给予小檗碱灌胃(300 mg/kg,1次/d),C组和D组给予等量生理盐水灌胃。监测B组小檗碱血药浓度动态变化和所有大鼠的体重及空腹血糖变化。第8周所有大鼠安乐死,Micro-CT扫描并三维重建左侧上颌骨,测量磨牙牙槽骨吸收值和颌骨微观骨形态学参数值;制备磨牙硬组织切片+抗酒石酸酸性磷酸酶染色(TRAP),对破骨细胞进行计数;采集右侧第一、二磨牙龈下菌斑,行16S rDNA测序和菌群多样性分析;取游离龈(20 mg),ELISA法检测TNF-α和IL-1β的含量。结果:①血药浓度分别在给药后第1 h和3 h达到峰值。②C组体重不断增加,D组和B组则逐渐减轻;③血糖含量、炎症因子含量、破骨细胞数量、牙槽骨吸收值比较,C组B组>D组(P<0.05);骨小梁间隙则是D组>B组>C组(P<0.05);⑤龈下菌斑OTUs分析和Shannon多样性分析:C组>B组>D组(P<0.05);⑥主坐标分析:C组和B组无显著分界,但均与D组间具有显著分界;⑦龈下菌群结构:C组多样性良好;B组多样性降低;D组趋向单一化。结论:小檗碱可有效降低糖尿病大鼠血糖含量、炎症因子水平和破骨细胞数目,改变颌骨微观结构,并减少牙槽骨的吸收。

Objective: To verify whether berberine is helpful to the treatment of periodontitis in diabetic rats. Methods:Thirty male Sprague-Dawley rats were randomly divided into 3 groups: control group(group C), berberine group(group B)and diabetes group(group D). Group B and D were administrated with high glucose diet and streptozotocin to induce diabetes model. On the third day after modeling, animals in group B was given berberine through intragastric gavage(300 mg/kg, once daily), While the rats in group C and group D were given the same amount of saline. The dynamic concentration of berberine in plasma of group B and the body weight and fasting blood glucose of all rats were monitored.All rats were euthanized in the 8th week, by Micro-CT scan and 3D reconstruction of the left maxilla, the alveolar bone resorption value of the molar area and the microscopic morphological parameters of the jaw bone were measured. The hard tissue sections of the molar were prepared for tartrate-resistant acid phosphatase(TRAP) staining, and numbers of osteoclasts were counted. The plaques from the first and second molars on right jaw were collected, for 16S rDNA sequencing and microbial diversity analysis. Free gingiva was disected(20 mg from each site) for detection of TNF-α and IL-1βby ELISA. Results: ①Blood concentration of berberine group peaked at 1 h and 3 h after administration. ②Weight of group C increased continuously, while that of group B and group D decreased gradually. ③Blood glucose, inflammatory factor, number of osteoclasts and alveolar bone resorption sort: group C < group B < group D( P <0.05). ④ The sort of trabecular bone thickness and trabecular bone number: group C> group B > group D(P<0.05);space between trabecular sort: group D> group B > group C(P<0.05). ⑤OTUs analysis and Shannon diversity analysis sort: group C > group B >group D(P<0.05). ⑥Principal coordinate analysis showed there was no significant demarcation between group C and group B, but there was a significant demarcation between group D and these two groups. ⑦The component of subgingival flora showed the diversity of group C was better than that of group B, while the diversity of group D tend to be lost.Conclusion: Berberine can reduce the blood glucose level, inflammatory factor level and number of osteoclasts in diabetic rats, change the microstructure of the jaw bone, and prevent the absorption of alveolar bone.