| Objective: Diabetes mellitus and periodontitis are the two most common chronic diseases affecting human health,and periodontitis is listed as the sixth complication of diabetes mellitus,and the prevalence of periodontitis in diabetic patients is three times that of non-diabetic patients.Meanwhile,the state of periodontitis also affects the control of blood sugar.Quercetin is a kind of natural flavonoids which exists widely in nature and has multiple biological activities.This study aims to through the establishment of diabetic periodontitis and periodontitis rats model,selection of diabetes periodontitis quercetin lavage treatment in rats,observe each periodontal tissue pathological changes,the alveolar bone absorption situation,comparing each weight,blood sugar levels,periodontal tissue and serum low factor 1 alpha(Hypoxia inducible factor 1 alpha,HIF-1α)and its downstream factor vascular endothelial growth factor(vascular endothelial growth factor,VEGF)change,To investigate the interaction between diabetes and periodontitis and the effect of quercetin on diabetic periodontitis in rats.Methods: 50 SD rats were randomly divided into 5 groups.The experiment was carried out after feeding in a constant temperature environment without special pathogenic bacteria for 1 week.The experimental groups were as follows: normal group(C group);Periodontitis group(P group);Diabetes group(D group);Diabetes + Periodontitis group(D+P group);DP+Q:Diabetes + Periodontitis + quercetin group(DP+Q group).Group C did not interfere with normal feeding.The rats in group P,DP and DP+Q were fed adaptively for one week.The neck of the left and right maxillary first molars of anesthetized rats were ligated with 2 orthodontic ligature wire.The ligature wire was placed in the gingival sulcus and knotted near the middle buccal side to establish the model of periodontitis.After ligation,the rats in group P were fed with sugar water(glucose 100g/L)moistened common rat diet.D group,DP group and DP+Q group were fed with high glucose and high fat diet.With high sugar and high fat feed 6 weeks group D,intraperitoneal injection of DP and DP+Q group chain urea with cephalosporins(STZ)(60 mg/kg)preparation diabetes model,induction and induction in 3 days before the fasting blood glucose in rats were measured,the measured blood sugar value >16.7 3 times in a row tendency for diabetes building success.The weight of rats in each group was measured as baseline weight.After the establishment of diabetic model,the rats in DP+Q group were given 150mg/kg quercetin by oral administration at 10:00 every morning for 4 weeks,and the other rats without quercetin were replaced by the same amount of normal saline.The general state of the rats was recorded during feeding.Four weeks later,the body weight and fasting blood glucose of the rats were measured in each group.The head was severed and the serum was stored after centrifugation.The maxilla was fixed in 4% parafor-maldehyde for 48 hours,then cut along the middle palatal suture,and the right side was decalcified in 10%EDTA for 6weeks to make continuous periodontal sections,and HE staining was performed to observe the periodontal inflam-mation in each group.Electronic vernier calipers were used to measure the amount of alveolar left bone absorption.The expression levels of HIF-1α and VEGF in serum of rats were detected by enzyme-linked immunosorbent assay.The expression level of HIF-1α in periodontal tissues of rats was detected by immunohistochemical staining.Results:1.The general state and histology of rats in each group were observed1.1 Normal group: The rats had normal metabolism,quick action and dry padding in the feeding process.The gross observation of gingival tissue before the death: gingival shape is good,a small amount of soft scale can be seen on the tooth surface,and periodontal probe basically has no bleeding;Histological observation: the gingival epithelium was intact without loss of attachment,no infiltration of inflammatory cells in connective tissue,and the alveolar crest was regular in shape.1.2 Periodontitis group:The rats had normal metabolism,quick action and dry padding in the feeding process.The gross observation of gingival tissue before the death: soft scale was observed on the tooth surface of the first molar on both sides of the upper jaw,a large amount of calculus and food residue were observed on the tooth neck,bleeding was observed on probing examination,and periodontal pockets were found in red and swollen gingival tissues.Histological observation: erosion of gingival epithelium,proliferation of epithelial nail process and proliferation to the root,inflammatory cell infiltration in connective tissue,osteoclast formation in alveolar bone,destruction of intrinsic alveolar bone.1.3 Diabetes group: During feeding,the rats showed symptoms of polydipsia,polyphagia and polyuria,emaciation,slow action and slow reaction,damp and peculiar smell of padding,and retinopathy in a few rats.The gross view of gingival tissue before the death: soft scale can be seen on the tooth surface,gingival is slightly red and swollen,and the probing examination shows slight bleeding,without deep periodontal pocket.Histological observation:the gingival epithelium was intact without loss of attachment,there were mild inflammatory cell infiltration in connective tissue,and the alveolar crest was regular in shape.1.4 Diabetes+Periodontitis group: During feeding,the rats showed symptoms of polydipsia,polypia and polyuria.Compared with group D,the rats were emaciated,slow in action and slow in reaction,and the bedding material was moist and with peculiar smell.Compared with group D,the rats in DP group were emaciated,slow in action and reaction,and the padding was moist with odor.Macroscopic observation of gingival tissue before the death: Soft scale could be seen on the tooth surface of the first molar on both sides of the rats,and a large amount of calculus and food residue could be seen on the tooth neck.It was easy to bleed when probing,and gingival was red and swollen with deep periodontal pockets.Histological observation: erosion of gingival epithelium,proliferation of epithelial nail process and proliferation to the root,a large number of inflammatory cell infiltration can be seen in the connective tissue,periodontal membrane fibers and collagen fibers are disordered,a large number of bone filling pits can be seen in the alveolar bone,and the height of alveolar ridge is significantly reduced.1.5 Diabetes + Periodontitis + quercetin group : During feeding,the symptoms of polydipsia and polypsia were improved.Compared with the DP group,the rats were stronger and more active,and the bedding material was damp with peculiar smell.Compared with the DP group,the rats in DP+Q group had stronger body shape,more active movement,and damp bedding with odor.Macroscopic observation of gingival tissue before the death : soft scale was observed on the dental surface of the first molar on both sides of the rats,a large amount of calculus and food residue were observed on the tooth neck,and bleeding was observed on exploratory examination.Compared with DP group,gingival redness and swelling were less severe,and periodontal pockets were formed.Histological observation: the gingival epithelium was relatively complete,there was mild inflammatory cell infiltration in the periodontal pocket,the periodontal membrane was continuous,the collagen fiber hyperplasia appeared,and the height of alveolar bone was reduced.2.Blood glucose level and weight test results2.1 Blood glucose monitoring Fasting blood glucose of rats in each group before STZ induction: C:3.5±0.16;P:3.6±0.18;D:3.6±0.16;DP:3.7±0.17;DP+Q:3.6±0.21。Fasting blood glucose of rats in each group before STZ induction: C:3.7±0.13;P:3.7±0.20;D:24.5±1.54;DP:25.9±1.86;DP+Q: 27.6±1.60。Blood glucose before death: C:3.7±0.18;P:3.8±0.15;D:25.3±1.44;DP:28.9±2.16;DP+Q: 27.4±2.01。Before induction,there was no significant difference in fasting blood glucose(P>0.05),after STZ induction,the fasting blood glucose of group D,group DP and group DP+Q was significantly different from that before STZ induction(P<0.05);Before death,the fasting blood glucose of rats in DP group was higher than that in D group(P<0.05),the fasting blood glucose of rats in DP group was higher than that in DP+Q group(P>0.05).2.2 Body weight monitoring Baseline weight:C:310±15.0;P:305±12.0;D:300±16.0DP:290±20.0;DP+Q:295±18.0。weight before death: C:320±18.0;P:311±13.0;D:230±14.0;DP:225±20.0;DP+Q:220±12.0。Before death,the body weight of rats in groups D,DP and DP+Q was significantly different from that in groups D,DP and DP+Q at baseline(P<0.05).The body weight of rats in DP group was significantly lower than that in P group(P<0.05).3.Alveolar bone resorption((?) ± s,mm)C:0.530±0.11;P:0.986±0.06;D:0.566±0.10;DP:1.146±0.20;DP+Q:0.846±0.21.Alveolar bone uptake in P and DP groups was significantly higher than that in C group(P<0.05);Alveolar bone uptake in DP group was higher than that in P group(P<0.05);The alveolar bone uptake in DP+Q group was significantly lower than that in DP group(P<0.05);The amount of alveolar bone uptake was similar between group C and group D,but no significant difference was found(P<0.05).4.Detection of serum HIF-1α and VEGF levels((?) ± s,mm)Serum HIF-1α content:C:1285.29±36.87;P:1438.63±37.39;D:1917.91±88.85;DP:2115.61±70.37;DP+Q: 1787.72±100.98.The serum HIF – 1α cont ent in groups P,D,DP and DP+Q was higher than that in group C(P<0.05);The serum HIF–1α content in DP+Q group was lower than that i n DP group(P<0.05);The serum content of D group and DP group was higher than that of P group(P<;0.05);Serum VEGF content:C:65.37±3.47;P:68.54±2.19;D:70.42±3.43;DP:75.92±3.60;DP+Q:73.25±3.54.The expression of VEGF was the lowest in group C and the highest in group DP.The DP+Q group was smaller tha n the DP group,and the difference was not statistically significant.Immu nohistochemical results of HIF-1α in periodontal membrane5.Immunohistochemical results of HIF-1α in periodontal membraneThe average optical density value of HIF-1α expression: C:0.110± 0.0063;P:0.135±0.0055;D: 0.189±0.0088;DP:0.221±0.0084;DP+Q: 0.147± 0.0085.Immunohistochemical detection of positive signal mainly appeared in the periodontal membrane cytoplasm or nucleus,only a few cells in C group showed weak positive expression of HIF-1α,significantly lower than the other groups(P<0.05);The positive expression of HIF-1α was strongest in DP group(P<0.05),the positive expression of HIF-1α in DP+Q group was lower than that in DP group(P<0.05).Conclusions:1.Ⅱ diabetes rats,periodontitis rats,Ⅱ diabetes periodontitis rats model success.2.There is an interaction between diabetes and periodontitis.Diabetes aggravates periodontal tissue inflammation,which also leads to the increase of fasting blood glucose and serum HIF-1α content in diabetic rats.3.Quercetin can relieve Ⅱ diabetes periodontitis periodontal tissue i nflammation in rats,and helps reduce HIF-1α content in serum and perio dontal membrane. |
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