车前子对晶状体氧化损伤所致LEC凋亡抑制作用的实验研究

目的　研究归肝经明目中药车前子对实验性晶状体氧化损伤所致晶体上皮细胞 (LEC)凋亡及其凋亡小体形成的影响。方法　将SD大鼠的透明晶状体温育在含有 3 0 0 μmol/L过氧化氢的培养液中复制LEC凋亡的模型 ,同时加入终浓度为 15 %的车前子水提液共同孵育 2 4h后 ,取晶状体前囊膜采用TUNEL法 (末端脱氧核苷酸转移酶介导的生物素化脱尿苷三磷酸末端标记 )检测LEC凋亡率 ;透射电子显微镜观察LEC超微结构改变和凋亡小体形成。结果　车前子可明显抑制LEC凋亡 ,且抑制能力均强于白内停。过氧化氢对照组LEC凋亡率 (92± 2 .5 5 )显著高于空白对照组 (3 .5± 1.84) ,(t =62 .97,P <0 .0 1) ;车前子组LEC凋亡率为 16± 2 .8与过氧化氢组有非常显著差异 (t=62 .2 6,P <0 .0 1)。透射电镜观察到过氧化氢组绝大多数LEC均处于细胞凋亡的各阶段。车前子组多数晶状体上皮细胞呈凋亡早期改变 ,少数晶状体上皮细胞呈凋亡改变 ,未见有凋亡后继发性坏死的LEC。结论　车前子可明显抑制LEC凋亡 ,其显著抑制LEC凋亡的作用可能是其防止和延缓白内障发生发展的细胞学机制。

Experimental Study on Semen Plantaginis Inhibiting LEC Apoptosis Induced by Oxidative Damage of Lens

Objective To study the influence of Semen plantaginis(SP) belonging to liver-meridian eyesight-improving traditional Chinese herbs on lens epithelial cell(LEC) apotosis and the formation of apoptosis body caused by oxidative damage.Method Lenses of Sprague-Dawley(SD) rats were incubated with 300μmol/l H 2O 2 to induce apoptosis model.The membrance of anterior lens capsule was taken with TUNET(Terminal deoxynucleotidy transferase mediated biotin-dUTP end label) method to determine LEC apoptosis rate after putting 15% final concentration of SP water extract in culture fluid and incubating for 24 hours.The ultrastrunctural change and apoptosis body formation of LEC were observed with transmission electron microscope.Results SP could obviously inhibit LEC apoptosis.Its effect was stronger than that of Baineiting group.LEC apoptosis rate in H 2O 2 control group(92±2.55) was significantly higher than that in blank control group(3.5±1.84)( t =62.97, P <0.01).There were significant differences between SP group(16±2.8) and H 2O 2 group( t =62.26, P <0.01).It was observed that most of LEC in H 2O 2 group were at different stages of apoptosis by electron microscope.Most of apotosis changes in LEC of SP were at early stage,a little of LEC appeared apoptosis change,there were no secondary necrosis of apoptosis.Conclusions SP can evidently inhibit LEC apoptosis,which may be the cellular and molecular mechanisms of SP preventing and delaying the development of the catarat.