| Objective:The aim of this study was to investigate the effects of calcitriol（1α,25-dihydroxyvitamin D3,1,25（OH）2D3,hereinafter referred to as 1,25D）on the inflammatory expression of gingival tissue in rats with type 2 diabetic periodontitis and the role of autophagy in this process.Methods:A total of 20 eight-week-old male SD rats（body weight 220±20 g）were selected,16 diabetes rats were fed with high fat combined with low dose streptozorcin（STZ injection）and 4 non-diabetes rats.The periodontitis model was established by silkcord combined with P.gingivalis.The rats was divided into non-diabetic blank group,diabetic blank group,diabetic 1,25D group,diabetic 3-MA group and diabetic1,25D+3-MA group,with 4 mice in each group.The solvent was prepared by propylene glycol:water:ethanol=60:30:10.The rats in different groups were respectively intraperitoneally injected with 0.2 ml of the solvent,1,25D at the ratio of 0.2μg/kg（1,25D/rat weight）in 0.2 ml solvent,3-MA at the ratio of 15μg/kg（3-MA/rat weight）in 0.2 ml solvent and 0.2μg/kg+3-MA every day for 1 week.The rats were executed after administration.We need to observe the following indicators:（1）Micro-CT was used to observe the alveolar bone loss in rats;（2）HE staining to observe the inflammation of tissue;（3）The expression of autophagy indexes LC3 and Beclin-1 and inflammatory indexes NFκB-p65 and TNF-αwere detected by immunohistochemical staining;（4）Western-Blot to detect the expression of LC3II/LC3I.Results:（1）The loss of alveolar bone in diabetic blank group was significantly higher than that in non-diabetic blank group（P﹤0.05）.（2）The expression of NFκB-p65 and TNF-αin gingival tissue of diabetic blank group was higher than that of non-diabetic blank group（P﹤0.05）.（3）The protein expressions of LC3,Beclin-1 and LC3II/LC3I in gingival tissue of diabetic blank group were lower than those of non-diabetic blank group（P﹤0.05）.（4）The expression levels of NFκB-p65,TNF-αand p-p65 in gingival tissue of diabetic 1,25D group were lower than those of diabetic blank group,and the expression levels of LC3,Beclin-1 and LC3II/LC3I were higher than those of diabetic blank group（P﹤0.05）.（5）The expression levels of NFκB-p65,TNF-αand p-p65protein in diabetic autophagy inhibitor 3-MA group were higher than those in diabetic1,25D group,and the staining intensity of LC3,Beclin-1 and the protein level of LC3II/LC3I in diabetic 1,25D group were lower than those in diabetic 1,25D group（P﹤0.05）.（6）The expression levels of NFκB-p65,TNF-αand p-p65 protein in gingival tissue of diabetic patients with 1,25D+autophagy inhibitor 3-MA were lower than those of diabetic patients with autophagy inhibitor 3-MA.The expression levels of LC3,Beclin-1 and LC3II/LC3I protein were higher than those of diabetic autophagy inhibitor3-MA group（P﹤0.05）.Conclusions:（1）In the type 2 diabetic periodontitis model,diabetic rats show increased inflammation;（2）The level of autophagy in type 2 diabetic rats is insufficient;（3）The inhibition of autophagy can aggravate the inflammatory response of diabetic rats;（4）1,25D inhibits the inflammatory response of gingival tissue in type 2 diabetic rats by promoting autophagy. |
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