Response Of H₂O₂ Stimulated Human Periodontal Ligament Cells To Baicalin

Oxidative stress (OS) is characterized by an accumulation of oxygen species (ROS) and play a key role in the progression of inflammatory diseases.We hypothesize that inflammatory events induce oxidative stress in the periodontal ligament cells during clinical periodontitis. Some studies provided evidence for benefit effects, but the links between certain antioxidant species and pathogenic mechanisms are still missing[6] Oxidative stress is gaining interest as an important factor in the etiology and pathogenesis of different dental disease including periodontal or mucosal inflammation, tumor development, or inflammation processes like periodontitis. Persistent and prolonged oxidative stress causes various pathological disorders such as stroke, heart attack, aging, and several degenerative diseases[7] ROS can be also produced continuously by periodontal-pathogenic bacteria or their by-products. ROS causes oxidative damage of periodontal tissue and cells and eventually mediates periodontitis[8]. Hydrogen peroxide (H2O2) is generated by almost all sources of oxidative stress and can penetrate cellular membranes[9].Baicalin is bioactive flavonoid compound extracted from the dried root of Scutellaria baicalensis Georgi (common name in China:Huangcen; a traditional Chinese medicine).It has been demonstrated to possess many pharmacological properties such as antimicrobial and anti-inflammatory activities[101.In addition, it has been demonstrated to significantly enhance the synthesis of both collagen and total protein in gingival fibroblasts[11]. Thus, in the present work we evaluated whether baicalin exert a modulatory effect on continuously provided H2O2 in primary cell cultures of hPDLCs.1 Objective:1.1 Use H2O2 for establishing damage modle of human periodontal ligament cells (hPDLCs)1.2 To investigate the protective effect of baicalin oxidative damage of hPDLCs after injury of H2O2.2 Methods:2.1 The original training and identification of hPDLCs.2.2 Using hPDLCs, cytotoxicity and H2O2 damage model were measured by the Cell Counting Kit-8 (CCK-8) assay.2.3 Hoechst33342 fluorescet staining method was used to observe the shape after injurying.2.4 FCM (flow cytometry method) detectcted cells apoptosis rate, contrastive analysis of incubated baicalin groups.2.5 LDH and MDA was texted intracellular and extracellular of each hPDLCs groups.3 Results:3.1 The cell proliferation activity decline in by the H2O2 concentration of 200μmol/L processing 2h,the OD value differences with others(P<0.05).3.2 The apoptosis rate increased by the H2O2 concentration of 200μmol/L processing 2h, the apoptosis rate differences with others (P<0.05).3.3 The H2O2 damaged cells treated by different concentrations of baicalin for 24h, 1-10μg/ml can effectively decrease the rate of early and lately apoptosis than others (P<0.05).3.4 The H2O2 damaged cells treated by different concentrations of baicalin for 24h,1 μg/ml can effectively decrease the intracellular and extracellular LDH、MDA contents & leakage rates than others (P<0.05).4 Conclusion:4.1 H2O2 (200μmol/L,2h) renovated oxidative stress damage on human periodontal ligament cells.4.2 Baicalin at a concentration of 1μg/ml renovated the apoptosis in on human periodontal ligament cells.