| Objective: To evaluate the extent of periodontal damage in type Ⅱdiabetic rats by the intervention with coenzyme Q10 on the established type Ⅱ diabetic rat model. Test the matrix metalloproteinases(MMP-9) in the rat periodontal tissue by immunohistochemistry, observe the changes of expression of the rat serum osteoprotegerin(OPG) and receptor activator for nuclear factor-κB ligand(RANKL) through the ELISA. To explore the effect of coenzyme Q10 in the course of periodontitis of diabetic rats. To provide theoretical reference of coenzyme Q10 in clinic adjuvant treatment or prevention of diabetes periodontitis.Method: One hundred and twenty healthy male Wister rats which weigh about 180 g were divided into 6 groups randomly, and 20 rats in each group: normal group(O group),periodontitis group(P group),diabetes mellitus group(DM group),diabetes mellitus periodontitis group(DMP group),coenzyme Q10 on diabetes mellitus group(QDMP). The animals were housed separately, feeding and water intake freely. Weigh all the rats, monitor the concentration of blood glucose in rat tail veins.1 Establishment of rat model of type Ⅱ diabetes mellitusAfter DM,DMP,QDM,QDMP group had been feed with high-sucrose-fat diet for 4 weeks, low-dose STZ was injected into abdomen to establish rat model of type Ⅱ diabetes mellitus. Measure the blood glucose after one to two weeks’ injection, the rats whose fasting blood-glucose(FBG) reach 16.65 mol/L were identified as the diabetes mellitus rats.2 Feed Coenzyme Q10 solutions.Rats in QDM,QDMP groups were fed lavage daily with 10% water- soluble coenzyme Q10 in accordance with 9mg/kg, other groups were fed with the same dose of normal saline. Prepare to establish periodon- titis,diabetes mellitus rats model after perfusion of coenzyme Q10 for 5 weeks. Feed Coenzyme Q10 solutions till the end of the experiment.3 Establishment of rats model of periodontitisThe P,DMP,QDMP groups were replicated rat priodontitis model by silk ligation and drank the water with dung from themselves. Observe the changes of rats in six groups throughout the experiment procedure including activity, diet, urine volume. Check( once every two days) the oral gingival tissue on bleeding, smelling, erosion and periodontal pocket formation. According to the periodontal indexes and X-ray, compare the degree of periodontal destruction of P, DMP, QDMP group. The mean depth of periodontal pocket and total attachment loss of rats in each group were measured and analyzed statistically before administered(setting up the diabetes mellitus periodontitis rats model successfully) and 5th, 9th(4th weekends of rats’ periodontal ligation), 11th(6th weekends of rats’ periodontal ligation), 13 th weekends(8th weekends of rats’ periodontal ligation) after administered. Rats in each group were given excessive anesthesia and open chest quickly to take blood, 3500r/m centrifuge for 10 minutes, reserve the upper serum, after setting up the diabetes mellitus periodontitis rats model successfully, the 5th weekend after administrated, 4th, 6th, 8th weekends of rats’ periodontal ligation. Periodontal tissues of mandible of maxillary first permanent molar of rats were taken rapidly and placed 40g/ L paraformaldehyde.(1)The gum tissues were dehydrated, embedded in paraffin. Paraffin slices were cut, immunohistochemical stained after antigen retrieval, observed by microscopy. Then the inflammatory cytokine MMP-9 of periodontal tissue was measured.(2)The ELISA was used to detect the expression of OPG and RANKL in rats’ serum.Results:1 Establishment of rat model of type Ⅱ diabetes mellitusThe rats in DM, DMP, QDM, QDMP groups will be models when their FBG is bigger than 16.65 mol/L.The comparison of blood glucose-level among every group: Record the six groups’ blood glucose level separately after model establishment, perfusion of coenzyme Q10 for 5 weeks, 4th, 6th, 8th weeks of rat’s ligation. The difference between diabetes mellitus group(DM group),diabetes mellitus periodontitis group(DMP group),coenzyme Q10 on diabetes mellitus group(QDMP) and normal group(O group),periodontitis group(P group)was statistical significant(P<0.05),and the difference between other groups has no statistical significance(P>0.05).2 Evaluate every groups’ periodontal status after periodontal ligation.The rats’ gingival tissue in O, DM, QDM groups didn’t appear obvious swollen gums and probe bleeding, no periodontal pocket formation and alveolar resorption. When the rats in P, DMP, QDMP had periodontal ligature: the rats in DMP group appeared swollen gums on the first odontoprisis in their underjaw and bleeded very easily when probing, and the periodontal pocket arised successively since the 2th weekend of rats’ periodontal ligature, the time is earlier than the QDMP and P groups, the periodontal pocket’s depth fortified faster than the QDMP group.The comparison between the probing depth and vertical alveolar bone absorption amount has no statistical significances(P>0.05) in the diabetic model establishment and the 5th weekend’s perfusion of Q10.The mean depth of periodontal pocket and total attachment loss on the first molar in the rats’ mandible at the 4th,6th,8th weekend of rats’ periodontal ligation: DMP group>QDMP group> P group. The measurement’s comparison between DMP, QDMP groups and O, DM, QDM groups was different at 4 weeks after ligation(P< 0.05), and the contrast between other groups has no statistical significance(P>0.05). The probing depth and alveolar bone absorption amount is greater than O, DM, QDM group, but P > 0.05, so it doesn’t have statistical significance.The differences between P, DMP, QDMP group and O, DM, QDM group were significant(P< 0.01) 6 weeks after periodontal ligation, the measurement is different between P group and OMP group(P<0.05). The differences between P, DMP, QDMP group and O, DM, QDM group were significant(P < 0.01) 8 weeks after periodontal ligation, the contrast between P group and DMP group, DMP group and QDMP group were statistically significant(P< 0.05).3 HE staining was observed in rat tissue morphologyO group, DM group, QDM rat gingival tissue very little inflammatory cell infiltration, collagen fibers arranged in order. 8 weeks after ligation, P group gingival epithelial hyperplasia in rats, lamina propria shows moderate inflammatory cell infiltration, fibroblast proliferation and collagen fiber bundles derangement.DMP group were significantly gingival epithelial hyperplasia seen a large number of inflammatory cells in the lamina propria infiltration, fibroblast proliferation and capillary proliferation, collagen fiber breakage most disintegration.QDMP rat gingival hyperplasia epithelium, lamina propria visible inflammatory cell infiltration, fibroblast proliferation and capillary proliferation, part of the cleavage of collagen fibers.4 The expression of MMP-9 in rat’s gingival tissue.The expression of MMP-9 in gingival tissue of rats in O group had no obvious change. In DM, DMP, QDM, QDMP group, the expressions were slightly higher than those of O, P group at the diabetic model establishment and the 5th weekend’s perfusion of coenzyme Q10, so it has statistical significance(P<0.05). The difference between DMP, QDMP group and O, P,DM, QDM group was statistically significant(P<0.05) at the 4th,6th,8th weekend of rats’ periodontal ligation, the contrast between P group and DMP group was statistically significant(P< 0.05),the DMP group and QDMP group were statistically significant(P< 0.05).5 The OPG, RANKL level in the rat’s serumTest the expression of OPG, RANKL in serum at 5 timings after the establishment of diabetic rat model, the difference between DM, DMP, QDM, QDMP group and P, O group was significant(P<0.01) The expression of OPG, RANKL enhanced after periodontal ligature, but the contrast between P group and DMP group was statistically significant(P<0.05). With the prolonged time of administration, the expression of OPG, RANKL in serum was significantly different at the 4th,6th,8th weekend of rats’ periodontal ligature(P<0.01).Compare the ratio of RANKL/OPG between O, P group and DM,DMP,QDM,QDMP group respectively after the diabetic model establishment, 5 weeks after the treatment, 4 weeks after periodontal ligation, 6 weeks after periodontal ligation, 8 weeks after periodontal ligation, the differences were significant(P<0.01). Compare DM, QDM group with QDM, DMP, QDMP group at 8th weekend of rats’ periodontal ligation, the difference is obvious(P<0.01).Conclusion:1 Build the rat model of type Ⅱdiabetes successfully by injecting low-dose STZ in rats’ abdominal. Successfully copied the rat periodontitis model through the silk ligation and drank the water with dung from themselves2 The study confirmed the negative regulation of diabetic periodontitis again by subintrant periodontitis of diabetic rats and aggravated damage of periodontal tissue.3 A dose of coenzyme Q10 may reduce the extent of the application of pathological periodontal tissue damage in diabetic rats periodontitis.4 The effect that the coenzyme Q10 on delaying or blocking the periodontitis of diabetic rats may be localized by rat periodontal tissue MMP-9, serum OPG, RANKL expression differences to achieve. |
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