Expression Change of P2X Receptors in Gingival Tissues of Experimental Periodontitis Rats
Objective To investigate the expression and changes of P2X receptors in gingival tissues of experimental periodontitis rats and to find out the P2X receptor subtypes related to the development of periodontitis.Methods 40 male SD rats were randomly divided into 4 groups(n=10 for each group):control group,2-week periodontitis group(2WP group),4-week periodontitis group(4WP group),and 6-week periodontitis group(6WP group).The experimental periodontitis rat model was established by ligating the bilateral maxillary first molars with 0.2 mm orthodontic ligation wire,and the gingival tissues of the rats in each group were collected for observation at 2,4,and 6 weeks after periodontal ligation.The periodontal conditions of rats in each group were evaluated using gingival index(GI),jaw Micro-CT scanning,and methylene blue staining.Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of P2X1-7 subtype receptors in gingival tissues of rats in each group;western blot and immunohistochemistry were employed to assess the protein expression levels of P2X7 receptors in gingival tissues of rats in each group;the changes in the concentrations of TNF-α and IL-1β in serum in each group were measured by ELISA.Results With the extension of periodontal ligation time,the degree of inflammation increased and the absorption of alveolar bone gradually increased(P＜0.01).The mRNA expression levels of P2X1,P2X2,P2X5,P2X6 and P2X7 receptors in gingival tissues of the 6WP group were significantly increased compared to those of normal rats(P＜0.05),whereas the mRNA expression of P2X4 receptors was markedly reduced(P＜0.05)and the mRNA of P2X3 receptors was almost not detected.Compared to the control group,the protein contents of P2X7 receptors in gingival tissues of each periodontitis model group were significantly increased(P＜0.05),and P2X7 receptors were mainly expressed in the oral epithelium of gingival tissues.Compared to the control group,the serum concentrations of TNF-α and IL-1β in each periodontitis model group were significantly elevated(P＜0.01),and the concentrations of TNF-α and IL-1β increased with the extension of ligation time(P＜0.05).Conclusion Significant changes in the mRNA expression of P2X1-7 subtype receptor were observed in gingival tissues of experimental periodontitis rats,among which the increased expression of P2X7 receptor was most prominent,suggesting that the P2X7 receptor in the gingival tissue might play an important role in the progression of periodontitis.
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