The Empirical Study About The Effect Of Youguiwan Prescription On The Periodontitisof Orchidectomizedrats With Osteoporosis

Objective: Animal model of osteoporosis and periodontitis was established through the male wistar rat orchidectomized and periodontal ligation. The influence of Youguiwan and Androgen to the experimental osteoporosis periodontitis and the bone metabolism was compared, to observed the therapy effection Youguiwan on experimental periodontitis in rats with osteoporosis. To explored the relations between masculine osteoporosis and the development of periodontitis, and provide experiment basis on clinical treats the masculine osteoporosis periodontitis.Method: Sixty 6-month-old male Wistar rats were randomly divided into six groups: normal group 10(i.e. group Normal), sham-operated periodontitis group 10(i.e. group Sham-PD), orchidectomy group 10(i.e. group ORX), orchidectomized periodontitis group 10(i.e. group ORX-PD), orchidectomized periodontitis with Youguiwan prescription therapy group 10(i.e. group YGT), orchidectomized periodontitis with Testosterone Undecamoate replacement therapy group 10(i.e. group ART). Group ORX, group ORX-PD, group YGW, group ART resection orchidectomy in rats, group Sham-PD only made a incision don't excise the testicle; 4 weeks after surgery, rats of group Sham-PD, group ORX-PD, group YGT, group ART were ligated bilaterally of their first maxillary molar teeth with 0.2mm steel-wire; after 3 months of orchidectomy (periodontal ligation 2 months), group ORX and group Normal were randomly selected and killed, take the rats'femur and maxillary, Make femur and maxillary tissue sedions and observe under light microscope, to identify osteoporosis model; Group Sham-PD and group Normal were randomly selected,killed,taken on jaw, the Maxillary First Molar tooth periodontal United biopsy was processed for histological observation, to identify periodontitis model. osteoporosis model and periodontitis model was established, group YGT rats were treated with Youguiwan prescription, according to the proportion of 5mg/kg body weight 2ml(equal to crude drugs 2g) daily gavage, group ART rats were treat with Testosterone Undecamoate, according to the proportion of 8mg/kg body weight match suspension with 2ml physiological saline, 1ml two times daily gavage;the remaining four groups was treated with oral administration of normal saline 2ml daily; after 3 months medication and rats were bled to death by femoral arteries bledding, the rats mandible, hemi-maxillary were immediately putted into 10% formaldehyde buffer solution for fixation, then by 10%EDTA decalcification solution, the production of dental Week biopsy were processed for HE staining to observe, light microscope observation; Tartate resistant acid phosphatase(TRAP) staining to observe osteoclast's quantity and distribution, and make statistics analysis processing; Immunohistochemisty was used to observe bone morphogenetic protein-2(BMP-2), observes BMP-2 the expression and makes statistics analysis processing. The other side of mandible, the check of the periodontal tissue between maxillary first molar and maxillary second molar, the manufacture organization refining, taking the supernatant after centrifugation, determining in gingival tissues the Alkaline phosphatase(ALP) the level, and makes statistics analysis processing.Results:1 Normal femoral and maxillary trabecular thickness, density were evaluated, Shum rules with the force direction; desexualization group manifested smaller femur trabecular bone, sparse, disorder and irregular. The results indicates flat desexualization method has been successfully in setting up animal models of osteoporosis.2 Histological observation, revealed gingival inflammation, periodontal ligament junctional epithelium vasodilator combination is not close, or has the proliferation to the root side.Indicate that periodontal ligation has been successfully set up animal model of periodontitis.3 Rat Periodontal Tissue Morphology HE staining, after 3 months medication, group Normal: junctional epithelium is located in cementum enamel boundary, gingival periodontal ligament fibers were arranged regularly, no absorption of alveolar crest.group Sham-PD: gingival epithelium has inflammatory cell infiltration,combined with epithelial proliferation or aversion to the root side, the formation of deep periodontal pocket,vasodilator hyperemia, alveolar crest was lower, absorption, and osteoclasts. Group ORX: junctional epithelium is located in cementum enamel boundary, with the tooth body union close, infiltrates not inflammatory, the osteoclast occasionally can be seen in the alveolar crest edge, but alveolar has no obvious resorption. Group ORX-PD: deep periodontal pocket was formatted, and some deep to apical 1/3, and some involved furcation, trench beneath the epithelium and lamina propria have a large number of inflammatory cell infiltration, blood vessel expansion of congestive exposed teeth alveolar bone and the surface has cubstantial absorption lacuna, more osteoclasts and less osteoblasts. Group YGT: periodontal pocket exist, but the bag is shallow, some involved furcation, trench beneath the epithelium and lamina propria have few inflammatory cell infiltration, at the alveolar ridge and cementum lacuna absorption edge more osteoblasts can be seen, more new bone formation can also be observed. Group ART: periodontal pocket exist, trench beneath the epithelium and lamina propria have a small amount of inflammatory cell infiltration,alveolar ridge and cementum lacuna absorption edge shows that osteoblasts existed, new bone formation can be seen.4 The change of the periodontal tissues of rat ALP level in every groups, group Sham-PD and group ORX were significantly higher than the group Normal (P<0.01), group ORX-PD was significantly higher than group Sham-PD and group ORX (P<0.05), group YGT and group ART were significantly lower than group ORX-PD (P<0.01), and group Normal had no significant difference (P>0.05), group YGT was significantly lower than group ART (P<0.01).5 Tartate resistant acid phosphatase(TRAP) dyeing determines osteoclast's distribution and the expression level in the periodontal tissues of rat, normal group: less osteoclast can be seen in the alveolar crest area, only individual osteoclast existence in the alveolar crest edge. Group Sham-PD: many osteoclast existences in the alveolar crest edge. Group ORX: In alveolar crest edge absorption dimple obviously few osteoclast existence. Group ORX-PD: in the alveolar crest area massive osteoclast existence. group YGT: in the alveolar crest edge osteoclast are rare. Group ART: in the alveolar crest edge osteoclast are few. The alveolar crest area osteoclast counting results, group Sham-PD was higher than the normal group significantly (P<0.01), group ORX-PD was higher than group Sham-PD obviously (P<0.01), group ORX and group Normal had no significant difference (P>0.05), group YGT and group ART were significantly lower than group ORX-PD (P<0.01), and group Normal had no significant difference (P>0.05).6 Immuno histochemistry determination of BMP-2 distribution of expression in the periodontal tissues of rat 's: BMP-2 mainly locates in the osteoblast, fibroblases the bone cell, normal group: The positive cell number are few, and, in the partial osteoblasts and fibroblasts to assume the weak positive expression, in the bone cell assumes the negative expression. Group Sham-PD, group ORX, group ORX-PD positive cell number increased, assumes positive expression. Group YGT and group ART positive cell number increases obviously, and in the osteoblast, fibroblasts and the partial bone cells assumes the strong positive expression. BMP-2 average light density value (OD) determination result, group Sham-PD, group ORX, group ORX-PD, group YGT, group ART obviously to be higher than the normal group (P<0.01); group YGT and group ART were significantly higher than group ORX-PD (P<0.01) And between two treatment groups there was no obvious difference (P>0.05).Conclusion: 1.Orchidectomized rats ligation of maxillary first molar successfully established periodontitis rat model of osteoporosis. 2. Osteoporosis, while the body also appeared jaw bone osteoporosis, and the osteoporosis may increase the severity of periodontal disease. 3.Youguiwan and androgen significantly reduced periodontal tissue osteoclast's expression quantity. 4.Significantly increased periodontal tissue BMP-2 expression, induces the bone, promoted mandibular bone new bone's formation that Youguiwan and Androgen have obvious therapeutic effect on periodontitis of osteoporosis. 5.Youguiwan may be reduces periodontal inflammation through down-requlating the periodontal tissue's ALP level.