硫化氢对氧应激下大鼠肝星状细胞的收缩和黏附作用

THE CONTRACTION AND ADHESION EFFECTS OF HYDROGEN SULFIDE ON OXIDATIVE STRESS OF HEPATIC STELLATE CELL IN RATS

目的探讨硫化氢(H2S)含量的变化对氧应激下大鼠肝星状细胞(HSC)收缩和黏附作用影响。方法将HSC分为空白组(B组,HSC-T6);对照组(C组,HSC-T6+500μmol/L FeN-TA);NaHS组(N组,N1:HSC-T6+500μmol/L FeNTA+100μmol/L NaHS;N2:HSC-T6+500μmol/L FeNTA+200μmol/L NaHS;N3:HSC-T6+500μmol/L FeNTA+300μmol/LNaHS;N4:HSC-T6 500μmol/L FeNTA+400μmol/LNaHS);GLBN组(G组,G1:HSC-T6+500μmol/L FeNTA+200μmol/LGLBN;G2:HSC-T6+500μmol/L FeNTA+400μmol/LGLBN)。应用500μmol/L次氮基三乙酸铁(FeNTA)制备氧应激模型。采用聚硅酮膜法直观检测氧应激模型大鼠肝星状细胞收缩情况、甲苯胺兰染色法测定细胞黏附率、Boyden Chamber小室法测定HSC跨膜迁移数量。给予不同浓度的外源性H2S供体NaHS和KATP通道阻断剂格列本脲(GLBN),在不同刺激条件下,观察上述指标。结果 HSC被NaHS处理后可见聚硅酮膜由于细胞收缩而引起明显皱纹,能够明显促进HSC收缩,而且随H2S浓度的升高和作用时间的延长,HSC收缩程度增强,这些作用能够被格列本脲阻止。加入200、400μmol/L H2S的HSC黏附抑制率分别为62.4%、81.7%,与对照组相比有显著差异;100、200、300、400μmol/L 4种不同浓度NaHS的HSC细胞迁移数量为6.43×103、8.85×103、11.34×103、14.86×103,与对照组(2.52×103)比较存在显著差异。结论 H2S在氧化应激下能促进HSC的收缩。H2S通过抑制细胞黏附、增加HSC跨膜迁移数量发挥细胞保护作用。

Objective To research the contraction and adhesion effects of Hydrogen sulfide on oxidative stress of hepatic stellate cell（HSC）in rats.Methods HSC were divided into four groups：Blank group（B group）;HSC-T6,Control group（C group）;HSC-T6＋500 μmol/LFeNTA,NaHSgroups（N group）;N1：HSC-T6＋500 μmol/LFeNTA＋100 μmol/LNaHS;N2：HSC-T6＋500 μmol/LFeNTA＋200 μmol/L NaHS;N3：HSC-T6＋500 μmol/LFeNTA＋300 μmol/LNaHS;N4： HSC-T6＋500 μmol/LFeNTA＋400 μmol/L NaHS.GLBN groups（G group）;G1：HSC-T6＋500 μmol/L FeNTA＋200 μmol/L GLBN;G2：HSC-T6＋500 μmol/LFeNTA＋400 μmol/LGLBN.Hepatic stellate cell with oxygen stress model in rats were prepared under the action of 500 μmol/L Fe-NTA,Use the method of assembling silicone m-embrane intuitive to detect the contract of hepatic stellate cells in rats.Armour aniline dyes blue was used to determine the cells aniline adhesion rate.The transmembrane migration numbers was determined by BoydenChamber.Different concentrations of exogenous H2S were given donor sulfur hydrogenated NaHS and KATP channels blocking agents of sodium blockers glyburide（GLBN）,under the condition of different stimuli,observe the indexes mentioned above.Results After HSC treated By NaHS,the silicone membrane to be found that cell shrinkage causing obvious wrinkles,can significantly promote HSC contraction,and with the H2S concentration increases and the time prolonged,the contraction degree of HSC enhanced.These effects can inhabit by glyburide.Join 200 μmol/H2S L and 400 μmol/L of adhesion,HSC inhibition rate were 62.4%,81.7%,respectively.compared with the control have significant difference.NaHS 100 μmol/L,200 μmol/L,300 μmol/L,400 μmol/L respectively were given to detect the HSC cell migration.The migration numbers were 6.43×103,8.85×103,11.34×103 and 14.86×103,respectively and compared with the control group 2.52×103 was significantly different.Conclusion H2S can promote the contraction of HSC in oxidative stress condition.HSC may plays a part to protect cell throuth inhibiting cell adhesion and increasing the number of HSC transmembrane migration.