| Objective: To observe the periodontal tissue morphology,blood glucose,alveolar bone resorption and the differences in the content of serum advanced glycation end products(AGEs)through establishing a periodontitis rat model,a diabetes rat model,a diabetic periodontitis rat model and diabetic periodontitis rats trested with quercetin.And explore the interaction between periodontitis and diabetes,and the therapeutic effect of quercetin on experimental diabetic periodontitis rats.Methods: Total seventy male SD rats weighing 130-150 g were randomly divided into 6 groups.There were 10 rats in the control group(C group)and the remaining 60 rats were randomly divided into 5 groups.They were periodontitis model group(P group),diabetes model group(D group),diabetic periodontitis model group(DP group),diabetic periodontitis non-treatment group(DP + N group),diabetic periodontitis quercetin treatment group(DP + Q group),there were 12 animals in each group.The C group did not interfere with normal feeding.Rats in group D were adaptively fed for one week.After feeding with high-sugar and high-fat diet for 6 weeks,a single intraperitoneal injection of 1% streptozotocin was injected at a dose of 50 mg / kg to induce a diabetic rat model.After 72 hours,fasting blood glucose was measured,and the blood glucose values were consecutively > 16.7 mmol / L for 3 consecutive times to obtain the successful modeling.After successful modeling,rats in D group continued to feed with high sugar and fat.After the adaptive feeding for one week,bilateral maxillary first molars were ligated with 0.2mm orthodontic ligature under anesthesia with 10% chloral hydrate in P,DP,DP + N and DP + Q group.Rats in P group were fed with ordinary food moistened with sugar water(glucose 100 g / L),and rats in DP group,DP + N group and DP + Q group were fed with high-sugar and high-fat rat food moistened with sugar water to induce a periodontitis model.Six weeks after ligation,one rat was randomly selected from each group to determine whether the periodontitis model was successful.Subsequently,rats in DP group,DP + C group and DP + Q group were injected with 1% streptozotocin according to the protocol of diabetic modeling.The rats in DP + Q group were gavaged with quercetin liposomes at 150 mg /(kg * d)every day at 10 am.In DP + N group,the same amount of blank liposomes were orally gavaged for 6 weeks.After 6 weeks,blood glucose values were measured,and femoral arterial blood was collected under anesthesia before the rats were sacrificed.Maxillary bone was taken off after sacrificing.Serum was collected by centrifugation of arterial blood,and AGEs contents was measured by ELISA.The maxillary bone was fixed in 4% paraformaldehyde for 48 hours,then cut along the zygomatic suture,and the right parts were decalcified in 10% EDTA for 6 weeks.HE staining was used to detect the morphological changes of the periodontal tissues.The left alveolar bone resorption was measured with vernier calipers after vertical peeling.Data were analyzed with statistical-software.Result: 1.Modeling results 1.1 Results of diabetes rat modelingAfter injection of streptozotocin,the symptoms gradually appeared,such as polydipsia,polydipsia and polyuria in the diabetes model group(D group).The dull hair and slow activity were appeared.The litter became dirty easily,daily replacement 2-3 times.And the blood glucose was always greater than 16.7 mmol / L.These showed successful diabetes modeling.1.2 Results of periodontitis rat modelingIn periodontitis model group(P group),rats were ligated with ligature for 6 weeks.After that,there was a large amount of calculus,plaque and food residues in the cervical surface of the first molars on both sides of the maxilla.The gum tissue was red and swollen with a deep periodontal pocket,the depth of the pocket exceeds 0.6mm,and the bleeding was detected.Teeth were loose(?°-?°).HE staining showed that the combined epithelium proliferated to the root,alveolar bone resorption,infiltration of inflammatory cells in the periodontal pockets,and edema degeneration of collagen,showing successful periodontitis modeling.1.3 Model of diabetic periodontitis ratRats in diabetic periodontitis model group(DP group),diabetic periodontitis non-treatment group(DP + N group),and diabetic periodontitis quercetin treatment group(DP + Q group)meet the above two results,proving periodontitis and diabetes model was induced successfully.2.After successful modeling,continued to feed for 6 weeks in each group.2.1 General condition of ratsThe rats in C group and P group were normal in metabolism,healthy in shape,shiny hair,quick in action,and the litter could be replaced once a day.In DP group,DP + N group,and D group rats,symptoms such as polydipsia,polydipsia,and polyuria aggravated,the body was thin,the hair was more dull,the action was slow,and the response was dull.Individual rats had retinopathy,and the litter was dirty with wet odor,needed to be changed 2-3 times a day.In DP + Q group,symptoms such as polydipsia,polydipsia and polyuria were improved.The body was thinner,the hair was dull,but the action was more active,the litter was moist and smelly,and it needed to be changed twice a day.2.2 Histological observationThe gingival epithelium of the rats in C group was intact,and the combined epithelium root was located at the enamel cementum.No adhesion loss was found.There was no obvious infiltration of inflammatory cells in connective tissue.The periodontal ligament fibers were arranged normally.No obvious bone resorption was observed.Gingival epithelium of rats in P group,DP and DP + N appeared erosion,combined with epithelium proliferation to the root,gingival epithelial spikes proliferated,a large number of inflammatory cells infiltrated under the epithelium and lamina propria,periodontal membrane fibrous and collagen fibers were edematous and deranged,and the arrangement was disordered.The alveolar bone height was significantly reduced,and a large number of bone resorption pits can be seen.Among them,the inflammation state of P group was lighter.The inflammatory cells were infiltrated,the periodontal ligament was continuous,and collagen fibers were proliferated.There were a large number of osteoblasts and new bone in alveolar bone.2.3 Blood glucose test results(± s,mmol/L)C group: 3.6 ± 0.16;P group: 3.7 ± 0.17;D group: 25.3 ± 1.44;DP group: 28.9 ± 2.16;DP + N group: 28.8 ± 2.25;DP + Q group: 20.6 ± 1.60.There was no statistically significant difference between C and P group;the blood glucose in D,DP,DP + N and DP + Q groups were significantly higher than those of C group(P <0.05);the blood glucose level in the DP + N group was higher than that in the DP + Q group(P <0.05).2.4 Alveolar bone resorption(± s,mm)After 6 weeks of treatment,the alveolar bone resorption was compared among groups.The alveolar bone resorption was more severe on the palatal side than on the buccal side,but the buccal mesial alveolar bone resorption was the most obvious.C group: 0.530 ± 0.11;P group: 0.986 ± 0.06;D group: 0.566 ± 0.10;DP group: 1.146 ± 0.20;DP + N group: 1.129 ± 0.21;DP + Q group: 0.846 ± 0.21.Alveolar bone resorption was significantly greater in P group,DP,and DP + N than in C group(P <0.05);the value of DP was greater than the value of P(P <0.05);the value of DP + N was greater than DP + Q(P < 0.05).2.5 Results of measurement of serum AGEs(± s,ng/L)C group: 100.3 ± 9.37;P group: 102.3 ± 3.20;D group: 115.3 ± 15.80;DP group: 128.8 ± 5.90;DP + N group: 127.6 ± 3.50;DP + Q group: 111.6 ± 4.33.The content of serum AGEs in D group,DP group,DP + N group and DP + Q group was significantly higher than that in C group(P <0.05);the values in P group and C were similar(P> 0.05);the value of DP group was higher than that in D group(P <0.05);the value of DP + N group was higher than DP + Q group(P <0.05).Conclusion:1.The periodontitis model,type 2 diabetes model and diabetic periodontitis model were successfully induced.2.Diabetes and periodontitis affect each other.Diabetes can lead to aggravated inflammation of periodontitis,and periodontal inflammation can increase blood glucose and serum AGEs in diabetic rats.3.Quercetin has a certain protective effect on the periodontal tissue of diabetic periodontitis rats,and helps reduce levels of blood sugar and serum AGEs. |
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