Research On The Effect Of BUSHENHUOXUEGUCHIFANG On The Periodontal OPG And RANKL Of Experimental Rats With Periodontitis

Objective: Using periodontal ligation combined with feeding sticky soft foods and high-sugar water to build rat periodontitis model, then treating experimental rats with periodontitis with BUSHENHUOXUEGUCHIFANG for four weeks and making tissue sections with specimens takenfrom periodontal ligament, which are undergone HE and immunohistochemical staining, so as to observe pathological and morphological changes thereby and their impacts on periodontal OPG and RANKL levels and research the functional mechanism of BUSHENHUOXUEGUCHIFANG in treating periodontitis.Methods: Randomly divide 24 male SD experimental rats into three groups: periodontitis group, periodontal treatment group, and control group. Anesthetizing experimental rats through peritoneal injection of pentobarbital sodium 0.3%(1ml/100g), peeling the gums, and then ligating the maxillary first molar with orthodontic stainless steel in diameter of 0.2mm. On the day after the operation, feeding them with soft foods and high-sugar water to build models of the periodontitis group and the periodontal treatment group. Rats of the control group were also anesthetized through peritoneal injection, but weren't undergone periodontal processing. They were fed with regular foods and water after the operation. Four weeks after the modeling, making sections of periodontal specimens from the first molars of two rats randomly selected from each group, and then indentifying whether the modeling was successful or not through HE staining. After the modeling was confirmed, removing the stainless steel ligatures of six rats from the periodontitis group and the periodontal treatment group. Treating rats of the periodontal treatment group with BUSHENHUOXUEGUCHIFANG for four weeks since the following day, while feeding rats of the periodontitis group and the control group with same amounts of normal saline. Four weeks later, 3 cutting periodontal tissue from maxillary first molar teeth of rats of the three groups, fixed with 4% neutral formalin, decalcified with 10% EDTA and embedded with paraffin wax, then making tissue sections. Applying HE staining to observe pathological and morphological changes of their periodontal tissues, and applying immunohistochemical staining to observe changes in mean OD value of OPG and RANKL. All data were analyzed and processed with SPSS19.0 statistical software.Results: 1 Four weeks after the model was built, the periodontal tissues of experimental rats in the periodontitis group and periodontal treatment group appeared inflamed, soft, and easy to bleed upon observation with naked eyes. The HE staining sections show that inflammatory cells of periodontal epitheliums increased, with capillaries enlargement, epithelium shift, periodontal pockets, osteoclasts increased or alveolar bone absorption. Upon observation with naked eyes, rats in the control group had no obvious periodontal inflammation and hard to bleed when touched. The HE staining sections show that their periodontal tissues were comparatively complete, without obvious inflammation, epithelium shift, osteoclasts or alveolar bone absorption; 2 Observe pathological and morphological changes of experimental rats' periodontal tissues through HE staining. The control group: Their periodontal tissues were comparatively complete, without obvious inflammation, epithelium shift, osteoclasts or alveolar bone absorption; The periodontitis group: Their periodontal tissues had obvious inflammation, with capillaries enlargement, epithelium shift, periodontal pockets, osteoclasts increased or alveolar bone absorption. The periodontal treatment group: Their inflammation was relieved, their periodontal epitheliums were recovered, periodontal pockets got better, and their osteoclasts were reduced, while their osteoblasts were increased; 3 Apply immunohistochemical staining to measure the mean OD value of RANKL. The mean OD value of RANKL of the periodontitis group was higher than that of the control group and the periodontal treatment group, despite the statistical error(P<0.05). No statistical difference existed between the control group and the periodontal treatment group(P>0.05); 4 Apply immunohistochemical staining to measure the mean OD value of OPG. The mean OD value of OPG of the periodontal treatment group was higher than that of the control group and the periodontitis group, despite the statistical error(P<0.05). No statistical difference existed between the control group and the periodontal treatment group(P>0.05).Conclusions:1 The animal periodontitis model can be successfully built through applying periodontal ligation while feeding sticky soft foods and high-sugar water.2 The BUSHENHUOXUEGUCHIFANG is effective in lowering RANKL expression in periodontal ligament.3 The BUSHENHUOXUEGUCHIFANG is effective in lowering OPG expression in periodontal ligament.4 The BUSHENHUOXUEGUCHIFANG can restrain the absorption of old bones, boost the growth of new bones, thus being effective for reconstruction of alveolar bones.