六味地黄丸对糖尿病伴牙周炎大鼠牙周组织中OPG与RANKL的影响

目的通过研究六味地黄丸对糖尿病伴牙周炎大鼠牙周组织中骨保护素(OPG)与核因子-κB受体活化子配体(RANKL)的影响,探索六味地黄丸对牙周炎治疗的作用机制。方法选用清洁级6周龄雄性SD大鼠30只,采用腹腔注射链脲佐菌素(STZ)诱导大鼠糖尿病模型,采用钢丝结扎法建立牙周炎模型。动物模型建立后,随机分成实验组和对照组,实验组进行六味地黄丸灌胃,对照组采用蒸馏水灌胃;并于灌药后8周处死动物,解剖上颌骨,制作3μm厚连续切片,荧光免疫组化检测牙周组织中RANKL、OPG表达。激光共聚焦显微镜(LSCM)采集图像,行灰度值分析,比较OPG和RANKL在糖尿病实验组和对照组大鼠牙周组织中的表达差异。结果灌药后8周,实验组的OPG灰度值比对照组增加,且差异有统计学意义(P0.05),而实验组RANKL灰度值比对照组明显降低,差异有统计学意义(P0.05)。结论六味地黄丸对糖尿病伴牙周炎大鼠的牙周组织炎症有明显抑制作用,六味地黄丸对牙周炎治疗机制可能是通过RANKL-细胞核因子κB受体活化因子(RANK)-OPG系统来实现。

Impacts of Liuwei Dihuang Pills on OPG and RANKL in Periodontal Tis- sue of the Rats with Diabetes Complicated with Periodontitis

Objective To study the impacts of Liuwei Dihuang Pills on OPG （osteoprotegerin）and RANKL（ receptor activator of nuclear factor - KB ligand）in periodontal tissue of the rats with diabetes com- plicated with periodontitis so as to explore the effective mechanism of Liuwei Dihuang Pills on periodontitis. Methods Thirty clean- grade,male SD rats of 6 weeks old were collected. The intraperitoneal injection of STZ （streptozotocin）was used to induce diabetes models in the rats. The ligation with steel wire was applied to set up periodontitis model. After modeling, the animal models were randomized into an experimental group and a control group. In the experimental group, the gastric infusion of Liuwei Dihuang Wan was used. In the con- trol group, the gastric infusion of distilled water was used. The animals were sacrificed in 8 weeks after infu- sion. The maxillary bone was dissected and prepared continuously as the slices 3 p.m in thickness. Fluores- cence immunohistochemical detection was adopted to determine the expressions of RANKL and OPG in perio- dontal tissue. The laser scanning confoeal microscope （LSCM）was used for image collection and grey level a- nalysis. The differences in RANKL and OPG expressions in periodontal tissue were compared between the ex- perimental group and the control group. Results In 8 weeks after gastric infusion, OPG grey value in the ex- perimental group was increased as compared with the control group,indicating the significant difference （P 〈 0.05 ）. RANKL grey value in the experimental group was reduced apparently as compared with the control group, indicating the significant difference （ P 〈 0.05 ）. Conclusion Liuwei Dihuang Pills has the obvious suppression on periodontal inflammatory symptom in the rats of diabetes complicated with periodontitis. The effective mechanism is achieved probably via RANKL- RNK- OPG system.