论文部分内容阅读
目的:探索黄岑苷对H_2O_2诱导人牙周膜细胞氧化损伤后的保护作用。方法:原代培养人牙周膜细胞(human periodontal ligament cells,hPDLCs)并鉴定。CCK-8法检测细胞增殖及活性,选择适合的H_2O_2浓度,建立人牙周膜细胞氧化损伤模型。采用Hoechst33342荧光染色法观察损伤后细胞核形态,同时应用流式细胞术检测各组人牙周膜细胞对H_2O_2诱导的细胞毒性反应变化。此外采用分光光度法检测各组细胞及其上清液的LDH、MDA含量,从而阐明黄岑苷对人牙周膜细胞氧化损伤后的保护作用。结果:CCK-8法显示在H_2O_2浓度为200μmol/L时,其光密度(optical density,OD)值与其他浓度组有差异(F=24.113,P=0.001);荧光染色法观察到细胞凋亡典型形态;流式细胞术显示1~10μg/m L的黄岑苷对H_2O_2处理引起的人牙周膜细胞损伤有保护作用;检测LDH、MDA含量和漏出率提示1μg/m L黄岑苷能减少H_2O_2损伤后细胞的氧化损伤产物及炎症产物。结论:适宜浓度的H_2O_2可导致人牙周膜细胞凋亡率增高,同时细胞内的LDH、MDA漏出率上升,造成氧化损伤,适当浓度的黄岑苷可以保护人牙周膜细胞经H_2O_2的氧化损伤。
Objective: To investigate the protective effect of baicalin against oxidative damage of human periodontal ligament cells induced by H 2 O 2. Methods: Primary cultured human periodontal ligament cells (hPDLCs) were identified and identified. CCK-8 method was used to detect cell proliferation and activity, and H 2 O 2 concentration was selected to establish a model of human periodontal ligament cell oxidative damage. Hoechst33342 staining was used to observe the morphological changes of nuclei. At the same time, the changes of cytotoxicity induced by H_2O_2 in human periodontal ligament cells were detected by flow cytometry. In addition, the contents of LDH and MDA in cells and supernatants of each group were detected by spectrophotometry, and then the protective effect of baicalin on the oxidative damage of human periodontal ligament cells was elucidated. Results: The CCK-8 method showed that the optical density (OD) value of H 2 O 2 was different from that of other concentrations (F = 24.113, P = 0.001) when the concentration of H 2 O 2 was 200 μmol / Flow cytometry showed that baicalin at the concentration of 1 ~ 10μg / mL had a protective effect on human periodontal ligament cells induced by H_2O_2 treatment. Detection of LDH, MDA content and leakage rate suggested that baicalin at 1μg / m L could reduce H_2O_2 Oxidative damage products and inflammatory products of cells after injury. CONCLUSION: H 2 O 2 with appropriate concentration can induce the apoptosis of human periodontal ligament cells and increase the leakage rate of LDH and MDA in the cells, resulting in oxidative damage. Appropriate concentration of baicalin can protect human periodontal ligament cells from H 2 O 2 oxidation damage.