Effect Of Glucocorticoids On Periodontal Tissue In Rats With Adriamycin-induced Nephropathy

ObjectiveEstablishment rat model of adriamycin nephrotic syndrome and by high-dose glucocorticoid intervention.Interleukin-6(IL-6),tumor necrosis factor-?(TNF-?)and subgingival Porphyromonas(P.g)were used to investigate the effects of periodontitis,nephritis and glucocorticoid application,to provide experimental evidence for oral treatment of patients with nephropathy.Sixty SPF healthy and male WISTAR rats were randomly divided into two groups: normal group,nephropathy group,30 rats in each group.The rats in the nephropathy group were treated with doxorubicin hydrochloride 4mg / kg at the interval of 2 weeks.The normal rats were injected with normal saline(4mg / kg).Four weeks after the second injection,30 rats were injected into the metabolic cage to collect 24 h urine,and the protein was detected by BCA protein quantitative kit,urine protein quantification> 100 mg / 24 h,the rat model of nephrotic syndrome was established successfully.After the success of the model,the rats of the two groups were sacrificed randomly and the kidneys were collected to make the pathological sections.The remaining 40 rats,20 rats in the normal group were randomly divided into blank control group,glucocorticoid group,10 rats in each group,nephropathy group 20 were divided into adriamycin nephropathy group,adriamycin nephropathy + hormone group,10 rats in each group.Glucocorticoid group and adriamycin nephropathy + hormone group were given methylprednisolone 30 mg /(kg · d)for 10 weeks,blank control group and adriamycin nephropathy group were given equal volume of normal saline.At the end of the intubation,the rats in each group were sacrificed and sampled:1.The levels of IL-6 and TNF-? in gingival crevicular fluid were measured by ELISA.2.The periodontal tissues of maxillary molars in rats were cut and make the sections of periodontal tissues.3.The post-subgingival plaque of rats was used to detect the detection rate and detection quantity of P.g in samples by using SYBR Green I real-time quantitative PCR(qRT-PCR)technique.Results1.IL-6 and TNF-? results:The levels of IL-6 and TNF-? in gingival crevicular fluid of glucocorticoid group,adriamycin nephropathy group and adriamycin-induced nephropathy group were higher than those in blank control group(P<0.05),The levels of IL-6 and TNF-? in adriamycin nephropathy + hormone group were significantly higher than those in the other three groups(P<0.01).The levels of IL-6 and TNF-? in glucocorticoid group and adriamycin nephropathy group no statistical difference(P>0.05).2.Observation of rat periodontal tissue sections by light microscope: the blank control group combined with the epithelium was located in the enamel dental bone,the periodontal ligament fibers of the gums were neatly arranged,no inflammatory reaction,no alveolar crest,adriamycin nephrotic group periodontal tissue only a small amount of inflammatory cell infiltration,periodontal fiber arrangement slightly disorder,glucocorticoid group ditch intraepithelial and lamina propria visible inflammatory cell infiltration,periodontal fiber disorder,the number of capillaries increased,expansion of congestion,adriamycin nephropathy + hormone gavage group of periodontal fibrous disorder,alveolar crest can be seen osteoclasts,relative to other groups of the most severe periodontitis.3.qRT-PCR result: The detection quantity of P.g in adriamycin nephropathy + hormone was higher than that in other three groups(P<0.05),and the detection quantity of P.g in adriamycin nephropathy group was higher than in control group and glucocorticoid group(P<0.05),there was no significant difference in the detection of P.g between the control group and the glucocorticoid group(P>0.05).The detection rate: the detection rate of P.g in blank control group and glucocorticoid group was 40% and 50% respectively.The detection rate of Pg in adriamycin nephropathy group and adriamycin nephropathy + hormone group was 90%,higher than that in control group and glucocorticoid group(P<0.05).Conclusions1.The rat model of this experiment was chronic kidney disease model,meet the requirements of this experiment,the manufacturing method is successful.2.Adriamycin nephropathy in rats with periodontal abnormalities,high-dose glucocorticoid treatment will increase the periodontal abnormalities,there is a correlation between periodontitis nephropathy and the use of glucocorticoid.3.Nephrotic patients should be regularly on the periodontal health check,used to guide the treatment and medication.