摘要
目的探讨吡诺克辛钠液对H2O2诱导的人晶状体上皮SRA01/04细胞凋亡的抑制作用。方法将体外培养的SRA01/04细胞分为3组:先加药组:先加吡诺克辛钠液300μL培养23.5h后再加0.5μmol·L-1 H2O2培养30min;后加药组:先加0.5μmol·L-1 H2O2,30min后再加吡诺克辛钠液300μL培养23.5h;对照组:不加药培养24h。对3组细胞标本进行一氧化氮合酶活性及羟自由基水平检测,以免疫组织化学法检测Bcl-2、Bax及Caspase-3的表达和TUNEL法检测凋亡率。结果后加药组的一氧化氮合酶活性(165.17±1.20)nmol·mg-1及羟自由基检测值(0.205±0.010)μmol·L-1均高于先加药组的(161.39±1.83)nmol·mg-1和(0.174±0.010)μmol·L-1,更高于对照组的(146.98±6.19)nmol·mg-1和(0.144±0.010)μmol·L-1,经统计学分析各组一氧化氮合酶活性和各组羟自由基检测水平差异有统计学意义(F=32.10,P<0.01;F=211.20,P<0.01);Bax及Caspase-3表达的灰度值后加药组为188.50±5.27和198.04±2.44,先加药组为154.02±7.74和186.58±2.40,对照组为142.36±3.33和157.48±1.21,3组差异均有统计学意义(均为P<0.01)。凋亡率后加药组为50%、先加药组为25%、对照组为5%,各组凋亡率通过χ2检验差异有统计学意义(χ2=103.98,P<0.01);而Bcl-2表达的灰度值对照组为150.05±9.60,先加药组为138.23±4.78、后加药组为126.67±0.59,经统计学分析各组扫描灰度值差异有统计学意义(F=14.22,P<0.01)。结论吡诺克辛钠液可通过减低氧化应激反应抑制H2O2诱导的细胞凋亡效应,且先加药组抑制效应大于后加药组,提示应用氧化应激抑制药对白内障可能具有一定预防效应。
Objective To explore the anti-apoptotic effects of Pirenoxine solu- tion on the lens epithelial SRA01/04 cells induced by H2O2. Methods The SRA01/04 ceils cultured in vitro were divided into three groups. A pre-drug group ：After co-culture of the SRA01/04 cells with 300 μL Pirenoxine solution for 23.5 hours,0.5 μmol · L^-1 H2O2 was added and cultivated for 30 minutes. Post-drug group：After 0. 5μmol · L^-1 H2O2 was added to the cultured SRA01/04 ceils for 30 minutes,the cells were co-culti- vated with 300 μL Pirenoxine solution for 23.5 hours. Control group：No any drug was added. In each group,the detection of the· OH radicle and NOS activity, immunocytochemistry of Bax, Bcl-2 or Caspase-3 expression and TUNEL assay were carried out respectively. Results The detected level of NOS activity （ 155.17 ± 1.20 ） nmol· mg ^-1 or · OH radicle （0.205 ±0.010） μmol · L^-1 in post-drug group were significantly higher than those in pre-drug group and control group, which in pre-drug group were （ 151.39 ± 1.83 ） nmol · mg ^- 1 and （ 0. 174 ± 0.010 ） μmol · L^-1 , respectively, in control group were （ 145.98 ±5.19）nmol · mg-1 and （0. 144 ±0. 010） μmol · L^-1 ,respectively. There were significant differences among different groups by one-factor analysis of variance（F=211.20,P 〈0.01 ;F= 32. 10,P 〈0.01 ）. The scanned gray-scaled means of Bax or Caspase-3 in post-drug group were 188.50 ± 5.27 and 198.04± 2.44, respectively, predrug group were 154. 02 ±7.74 and 185.58 ± 2.40, respectively, control group were 142. 36 ± 3.33 and 157.48 ±1.21 ,respectively,there were significant differences among different groups by one-factor analysis of variance（ all P 〈0.01 ）. The apoptotic rates in post-drug group,pre-drug group and control group were 50% ,25% and 5% , there was significant difference（χ2 = 103.98 ,P 〈0.01 ）. The scaImed gray-scaled means of Bcl-2 in control group,pre-drug group and post-drug group were 150.05 ±9.60,138.23 ± 4.78 and 126. 67 ± 0. 59,
出处
《眼科新进展》
CAS
北大核心
2014年第9期821-825,共5页
Recent Advances in Ophthalmology
