P38在体外培养鸡胚胎晶状体半乳糖性白内障形成中的作用

目的:探讨P38参与的晶状体上皮细胞凋亡的调节在体外培养的鸡胚胎晶状体半乳糖性白内障中的作用。方法:用30mmol/L半乳糖诱导体外培养的鸡胚胎晶状体建立半乳糖性白内障模型,在培养10d里,通过每日观察、照相、计算晶状体的混浊面积,观察MAPK-P38抑制剂SB203580对半乳糖引起的晶状体混浊程度的影响,并用TUNEL原位凋亡细胞染色方法,观察晶状体冰冻组织切片上细胞凋亡情况。结果:半乳糖30mmol/L能诱导体外培养的鸡胚胎晶状体产生周边皮质性混浊,随着半乳糖暴露时间的延长,晶状体的混浊面积增加,P38抑制剂SB203580能有效减轻半乳糖性晶状体的混浊。TUNEL检测显示,在培养2,5,10d的半乳糖性白内障晶状体切片上,凋亡细胞的百分数分别为1.7%,4.5%和12%,而SB203580处理的晶状体,凋亡细胞数明显减少,在相应点分别是0.7%,1.5%和1.4%。结论:P38可能通过参与晶状体上皮细胞凋亡调控在半乳糖性白内障的形成中起作用。抑制P38激活能阻止晶状体细胞凋亡,减轻白内障形成。

Attenuation of galactose cataract formation with inhibition of P38 activity in vitro

AIM:To determine the role of MAPK-P38 in development of galactose cataract by mediating cell apoptosis of chicken embryonic lenses in vitro.METHODS:The cataract model was developed by 30 mmol/L galactose in cultured chicken embryonic lenses.The lenses were grown for 10 days with or without inhibitor of P38,SB203580.The lens was observed and photographed daily,and the degree of opacification was quantified by using image analysis software.Apoptosis in the frozen section of cultured lenses was examined by TUNEL assay.RESULTS:The cultured lens developed opacification at the periphery cortex with 30mmol/L galactose,and the degree of opacification increased with time during the 10-day-culture period.SB203580-treated lenses developed similar cortical opacity,but the degree of opacification was attenuated.In the section of galactose cataract lenses,the percentage of apoptotic cells was 1.7%,4.5% and 12% at cultured day 2,5 and 10 respectively,but in SB203580-treated lenses it decreased dramatically to 0.7%,1.5% and 1.4% respectively.CONCLUSION:The activation of P38 may be involved in formation of galactose cataract by mediating the signaling pathway of lens epithelial cell apoptosis.Inhibition of P38 activity can prevent apoptosis of lens cells and attenuate cataract progress.