慈姑多糖对过氧化氢诱导的体外培养兔晶状体浑浊的改善作用

Ameliorative effect of sagittaria sagittifolia polysaccharide on rabbit lens opacity induced by hydrogen peroxide in vitro

目的:探讨慈姑多糖(SSP)对过氧化氢(H2O2)诱导的体外培养兔晶状体浑浊模型的改善作用及可能机制。方法:取透明兔晶状体随机分为对照组、模型组(200、400、800、1 600、3 200μmol/L H2O2)和干预组(0.5、1 mg/m L SSP)离体培养48、72 h,观察晶状体浑浊度变化,并利用试剂盒检测谷胱甘肽(GSH)和丙二醛(MDA)含量及超氧化物歧化酶(SOD)活力,Western Blot法检测晶状体中血红素加氧酶-1(HO-1)、肿瘤坏死因子-α(TNF-α)、B淋巴细胞瘤-2基因(Bcl-2)家族相关蛋白Bcl-2和Bax,以及半胱氨酸蛋白酶-3(Caspase-3)的蛋白表达。结果:与模型组比较,干预组有助于改善晶状体浑浊度,缓解云雾状浑浊形成,1 m g/m L SSP作用24 h保护效果最好;且与模型组比较,干预组中HO-1、Bcl-2蛋白表达及GSH、SOD含量或活力显著上调(P<0.01),MDA含量及TNF-α、Bax、Caspase-3的蛋白表达显著下调(P<0.01,P<0.05)。结论:SSP可以明显改善H2O2诱导的体外培养兔晶状体浑浊,可能与缓解晶状体氧化应激和炎症反应,抑制晶状体细胞凋亡有关。

Objective:To investigate the ameliorative effect and possible mechanism of sagittaria sagittifolia polysaccharide(SSP)on hydrogen peroxide(H,O,)-induced rabbit lens opacity in vitro.Methods:Transparent rabbit lenses were randomly divided into control group,H,02 model group(200,400,800,1600,3200μmol/L H,02),and intervention groups(0.5 and 1 mg/mL SSP).All the groups were cultured in vitro for 48 or 72 hours,then the changes of lens opacity were observed,the contents of glutathione(GSH)and malondialdehyde(MDA),and the activity of superoxide dismutase(SOD)were detected by kit,while the heme oxygenase-1(HO-1),tumor necrosis factor alpha(TNF-α),the B-cell lymphoma-2(Bcl-2)family related proteins including Bcl-2 and Bax,and cysteine protease-3(Caspase-3)were detected by Western Blot.Results:Compared with the model group,the SsP intervention group was helpful to improve the lens opacity and alleviate the formation of cloud haze.The protective effect of 1 mg/mL SSP for 24 hours was the best.And compared with the model group,the content or expression of HO-1,GSH,SOD and Bcl-2 in intervention group were significantly up-regulated(P<0.01),while the protein content of MDA,the protein expression of TNF-α,Bax and Caspase-3 were significantly down regulated(P<0.01,P<0.05).Conclusion:SSP can significantly improve the opacity of rabbit lens induced by H,O,in vitro,which may be related to alleviating oxidative stress and inflammatory reaction of lens and inhibiting apoptosis of lens cells.