Mechanism of Klotho's inhibition ofs H2O2-induced oxidative stress and apoptosis in periodontal ligament stem cells by regulating UCP2 expression
AIM:The aim of this study is to evaluate the levels of Klotho(Klotho protein)and uncoupling protein 2(UCP2)in the gingival crevicular fluid and gingival tissues of patients with chronic periodontitis,and to investigate the mechanism by which Klotho regulates UCP2 expression to inhibit H2O2-induced oxidative stress and cell apoptosis in periodontal ligament stem cells(PDLSCs).and to provide new therapeutic targets for the treatment of periodontitis.METHODS:Samples were collected from patients seeking treatment at the Changsha Stomatology Hospital.Gingival crevicular fluid(GCF)and periodontal ligament stem cells(PDLSCs)from patients with chronic periodontitis(n=12)and healthy controls(n=10)were divided into groups.The healthy control group PDLSCs were further divided into the H2O2 group,H2O2+Klotho group,H2O2+NAC group(positive control),and H2O2+DMSO group(blank control)according to the treatment.The concentration of Klotho in GCF of the periodontitis group was detected using ELISA,and the concentration of Klotho and the level of UCP2 in periodontal tissues of periodontitis patients were evaluated using Western Blot.The oxidative stress level was assessed by determining measuring the expression levels of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in the four groups of PDLSCs according to Western Blot.Cell apoptosis and necrosis were evaluated by PI staining and TUNEL assay to detect the expression levels of apoptosis-related proteins such as Caspase-3,BAX,Bel,MLKL,RIP1,and R1P3.Finally,rescue analysis was conducted by inhibiting UCP2 expression.RESULTS:Klotho showed relatively low levels in GCF and gingival tissues of patients with chronic periodontitis.The UCP2 levels in gingival tissues of patients with chronic periodontitis were significantly lower compared to the healthy control group.The analysis of the correlation between Klotho and UCP2 expression in periodontitis patients and healthy controls revealed a positive correlation.In the healthy control group,UCP2 expression was downregulated in PDLSCs treated with H2O2 alone,while UCP2 expression was upregulated in PDLSCs treated with Klotho+H2O2.This indicates that Klotho inhibits oxidative stress and cell apoptosis induced by H2O2 in PDLSCs.The rescue analysis showed that knockdown of the UCP2 gene can inhibit the effects of Klotho on PDLSCs.CONCLUSION:The expression of Klotho and UCP2 were positively correlated,and Klotho inhibited oxidative stress and cell apoptosis in PDLSCs induced by H2O2 by regulating the expression of UCP2.
NETL
NSTL
万方数据
