黄岑苷对H2O2诱导人牙周膜细胞损伤后作用的研究

Response of H_2O_2 stimulated human periodontal ligament cells to baicalin

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 responses of baicalin through oxidative damage of human periodontal ligament cells（h PDLCs）after exposed to hydrogen perioxide（H2O2）. Methods：The h PDLCs were cultured from healthy adult premolar teeth. Cell counting kit-8（CCK-8）was used to detect cell proliferation and activities. Appropriate concentration and time of H2O2 was chosen to establish the model. Hoechst33342 fluorescent staining was used to observe the cell nucleus of apoptosis. Fluorescent staining technique was combined with cytometry to investigate the influence of baicalin on the apoptosis and necrosis of H2O2 exposed cells. The leakage rates of LDH and MDA were calculated by spectrometer thus to elucidate the protective effect of baicalin on human periodontal ligament cells.Results：Results of CCK-8 showed that at a concentration of 200 μmol/L,the optical density of H2O2 was different from other groups（F=24.113,P=0.001）. The typical cell nucleus of apoptosis was revealed by fluorescent staining. Flow cytometry showed that 1 to 10μg/m L baicalin can protect h PDLCs exposed to H2O2. At the concentration of 1 μg/m L,baicalin can decrease the amount of the total cell LDH MDA leakage rates than the model group cells,even than the other concentration groups. Conclusion：H2O2at the concentration of 200 μmol/L renovate oxidative damage on h PDLCs. Baicalin at a concentration of 1 μg/m L protect h PDLCs of H2O2 damage.