1.High glucose exacerbates the inflammatory response in gingival fibroblasts through oxidative stress and mitochondrial DNA release
GENG Yiran ; ZANG Xiaoying ; LIU Jia ; LUAN Qingxian
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(12):1030-1040
Objective:
To investigate if high glucose (HG) exacerbates Porphyromonas gingivalis (P.g) lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs) and to explore the underlying mechanisms. To provide a basis for the mechanism of diabetes aggravating periodontitis.
Methods:
HGFs were divided into four groups: the control group (basal medium), the LPS group (treated with 5 μg/mL P.g-LPS for 24 h), the HG group (treated with 25 mmol/L glucose for 24 h), and the HG+LPS group (treated with 25 mmol/L glucose + 5 μg/mL P.g-LPS for 24 h). After culturing for 24 h in the respective media, the cells were harvested for experiments. Intracellular reactive oxygen species (ROS) and mitochondrial reactive oxygen species (mtROS) were detected using 2 ', 7' - dichlorodihydrofluorescein diacetate (DCFH-DA) and MitoSOX Red staining, respectively. Fluorescence intensity was analyzed by confocal fluorescence microscopy and directly measured in cell suspension. Immunofluorescence was used to detect changes in mitochondrial DNA (mtDNA) content of HGFs. Real-time fluorescence quantitative PCR was used to detect the content of mtDNA in cytoplasm and cell supernatant. Protein expression of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway was assessed by western blot, while mRNA expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were detected by PCR.
Results:
Compared to the control group, both the LPS group and the HG group exhibited a significant increase in ROS and mtROS, with a more pronounced elevation in the HG+LPS group, demonstrating a synergistic effect (ROS: F = 396.5, P < 0.001; mtROS: F = 29.38, P < 0.001, CI < 1). The cytoplasmic mtDNA content was significantly elevated in the LPS group, with a more marked increase in the HG+LPS group (F = 27.85, P < 0.001). The supernatant mtDNA levels were significantly higher in both the LPS and HG groups, with a more pronounced elevation in the HG+LPS group (F = 15.26, P < 0.001). The phosphorylated proteins p-STING, p-TBK1, and p-P65 in the cGAS-STING pathway showed varying degrees of activation in the LPS and HG groups, reaching the highest levels in the HG+LPS group (p-STING: F = 52.67, P < 0.001; p-TBK1: F = 15.67, P = 0.001; p-P65: F = 9.83, P = 0.005), while p-IRF3 showed no significant differences among the groups (P = 0.072). Pro-inflammatory cytokine TNF-α was significantly higher in the HG+LPS group compared to the control group (F = 15.05, P < 0.001), and IL-1β increased in both the LPS and HG groups, with a more pronounced rise in the HG+LPS group (F = 30.98, P < 0.001). IL-6 showed no significant differences among the groups (P = 0.847).
Conclusion
High glucose and LPS act synergistically to enhance oxidative stress, accompanied by increased mtDNA release, which activates the cGAS-STING pathway, thereby amplifying the inflammatory response in HGFs.
2.Rotational displacement on the sagittal plane in unstable pelvic fractures: a clinical case study
Shenglu CAO ; Yiran WANG ; Kai FENG ; Wei LING ; Geng PENG ; Gang WANG
Chinese Journal of Orthopaedic Trauma 2016;18(2):115-119
Objective To investigate the clinical significance and distribution of different rotational displacements on the sagittal plane in unstable pelvic fractures radiologically.Methods We retrospectively reviewed 82 patients with unilateral pelvic fracture who had been treated at our department from June 2005 to June 2015.They were 48 men and 34 women,with an average age of 43 ± 18 years.By Tile classification,there were 48 cases of type B and 34 cases of type C.Their rotational displacements on the sagittal plane of the pelvic ring were determined on the anteroposterior X-ray films of the pelvis by measuring the changed distances from the top of iliac crest to the lowest point of ischial tuberosity and to the pubic tubercle.The data were recorded for analysis of different displacements and their clinical significance.Results Of all the 82 patients,based on the preoperative radiographic evidence,36 (43.9%) showed sagittal plane rotational displacement.Of them,28 (77.8%) showed supination displacement and 8 (22.2%) pronation displacement.The preoperative CT three-dimensional reconstruction confirmed the findings from the X-ray films.Twenty-one of the 48 cases of Tile type B (43.8%) and 15 of the 34 cases of Tile type C (44.1%) showed sagittal plane rotational displacement.The success rate of closed reduction was,respectively,66.7% (24/36) in the 36 cases with sagittal plane rotational displacement,62.5% (5/8) in those with pronation displacement and 67.9% (19/28) in those with supination displacement,all significantly lower than the total success rate of closed reduction in this cohort(84.1%).Conclusions The sagittal plane rotational (pronation and supination) displacement of pelvic fracture can be determined by measuring special points and lines on the anteroposterior radiographs of the pelvis.The supination displacement on the sagittal plane is more common.The unstable pelvic fracture with sagittal plane rotational displacement is more difficult to reduce.


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