摘要
目的通过菊花影响过氧化氢(H_2O_2)氧化损伤的晶状体上皮细胞(LEC)内钙(Ca^(2+))、环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)信号转导的实验研究,探讨归肝经明目中药防治白内障的细胞和分子机制。方法将传代培养的牛LEC与H_2O_2共同孵育复制氧化损伤模型,同时加入菊花水提取液孵育后,四甲基偶氮唑蓝法(MTT)检测LEC活性、荧光分光光度法检测LEC内的钙离子浓度([Ca^(2+)]i)、放射免疫分析法检测LEC内cAMP和cGMP含量,探讨不同浓度菊花及孵育12 h、24 h和36 h的影响。结果①H_2O_2组LEC活性下降,菊花使氧化损伤的LEC活性显著增高(P<0.01),呈浓度依赖关系和时间依赖关系。②H_2O_2组LEC的[Ca^(2+)]_i升高,菊花使氧化损伤所致的LEC的[Ca^(2+)]_i显著降低(P<0.01),并呈时间-效应关系。③H_2O_2组LEC的cAMP浓度升高、cGMP浓度下降,菊花可使氧化损伤的LEC的cAMP水平显著降低(P<0.01)、cGMP水平显著升高(P<0.01),也呈时间-效应关系。结论通过[Ca^(2+)]_i、cAMP和cGMP信号转导系统及其相互作用调节生物学效应,可能是菊花防护LEC氧化损伤及减少细胞凋亡的细胞和分子生物学机制。
Objective To investigate the prevention of Flos Chrysanshemi （FC） on lens epithelial cell （LEC） damaged by oxidative stress and the mechanism of signal transduction. Methods Sub-cuhured bovine LEC were incubated with hydrogen peroxide （H2O2） and deferent concentrations of FC （0.8 mg/ml, 0.4 mg/ml, 0.2 mg/ml） for 12 h, 24 h and 36 h respectively. The LEC activities were observed by methyl thiazolyl tetrazolium assay （M3T）. LEC [Ca^2＋]i were detected by spectrofluorophotometer, intracellular cyclic adenosine monophosphate （cAMP） and cyclic guanine monophosphate （cGMP） of LEC were determined by radioimmunoassay. Results ① The activities of LEC increased in deferent concentrations of FC groups compared with H2O2 group （P〈0.01）, and showed time-dependent and dose-dependent relationship. ② The [Ca^2＋]i of LEC decreased in FC groups compared with H2O2 group （P〈0.01）, showed time-dependent. ③ The LEC cAMP decreased and the LEC cGMP increased in FC groups compared with H2O2 group （P〈0.01）, showed time-dependent. Conclusion ① FC may protect LEC against oxidative damages and increase activity of LEC. ② The calcium, cAMP and cGMP signal transduction system and the cross-talk may be the mechanisms of preventing and delaying cataract formation of FC.
出处
《福建中医学院学报》
2008年第6期26-29,共4页
Journal of Fujian College of Traditional Chinese Medicine
基金
福建省医药卫生重点项目(W2002505)
