中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2013年
9期
817-821
,共5页
徐国兴%崔丽金%林媛%吴雅冰
徐國興%崔麗金%林媛%吳雅冰
서국흥%최려금%림원%오아빙
石决明%晶体%上皮细胞%细胞,培养的%超氧化物歧化酶
石決明%晶體%上皮細胞%細胞,培養的%超氧化物歧化酶
석결명%정체%상피세포%세포,배양적%초양화물기화매
Concha haliotidis%Oxidative damage%Superoxide dismutase (SOD)%Glutathione (GSH)%Malondialdehyde(MDA)
目的 研究石决明提取液对氧化损伤的人晶状体上皮细胞(HLEC)的作用.方法 实验研究.采用双氧水(H2O2)干预体外培养HLEC氧化损伤模型,同时加入不同浓度(0.001％、0.01％、0.1％、0.3％)的石决明提取液,分成空白对照组、阳性对照组即H2O2组和不同浓度的石决明干预组,于1、3、5 d时用CCK-8检测各组HLEC的活力,倒置相差显微镜下观察细胞增殖和形态改变,3d后用化学比色法检测超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)和丙二醛(MDA)的水平.采用单因素方差分析进行样本均数间的多重比较,组间两两比较采用LSD-t检验.结果 不同时间点时各实验组间HLEC细胞活力的吸光度(A)值活力存在变化,第1、3、5天各实验组间HLEC细胞活力的A值分别是:空白对照组0.88、1.28、1.32；阳性对照组0.73、1.02、1.06；0.001％石决明提取液组0.73、1.03、1.06;0.01％石决明提取液组0.76、1.10、1.13;0.1％石决明提取液组0.79、1.22、1.21；0.3％石决明提取液组0.79、1.21、1.21；组间比较差异具有统计学意义(1d时F=23 922.42,p =0.05;3 d时,F=120 605.86,P＜0.05；5d时,F=150 939.45,P＜0.05).H2O2使细胞的活力降低,石决明提取液提高其活力,且在一定的时间和浓度范围内具有依赖性,其中第3天时0.1％石决明提取液组HLEC细胞活力的A值为最高(1.22).H2O2组损伤后,HLEC中SOD、GSH的水平降低(SOD 158.05 U/mgprot、GSH 15.05 mg/1000 mgprot)、而MDA量增高到18.11 nmol/mgprot,石决明提取液组使氧化损伤所致HLEC中抗氧化水平提高(SOD 188.64 U/mgprot、GSH 21.05 mg/ 1000 mgprot),脂质过氧化物升高有下降(MDA 14.16 nmol/mgprot),组间比较差异均具有统计学意义(P＜0.05).改变具有统计学意义(SOD:F=983.04,P＜0.05; GSH:F =444.44,P＜0.05; MDA:F=830.52,P＜0.05);阳性对照组中可见细胞核染色质浓缩、聚集,石决明组中细胞染色质的聚集有不同程度的减轻.结论 石决明提取液对体外培养HLEC氧化损伤具有保护作用,可提高细胞内抗氧化水平,减少有害产物的生成.
目的 研究石決明提取液對氧化損傷的人晶狀體上皮細胞(HLEC)的作用.方法 實驗研究.採用雙氧水(H2O2)榦預體外培養HLEC氧化損傷模型,同時加入不同濃度(0.001％、0.01％、0.1％、0.3％)的石決明提取液,分成空白對照組、暘性對照組即H2O2組和不同濃度的石決明榦預組,于1、3、5 d時用CCK-8檢測各組HLEC的活力,倒置相差顯微鏡下觀察細胞增殖和形態改變,3d後用化學比色法檢測超氧化物歧化酶(SOD)、還原型穀胱甘肽(GSH)和丙二醛(MDA)的水平.採用單因素方差分析進行樣本均數間的多重比較,組間兩兩比較採用LSD-t檢驗.結果 不同時間點時各實驗組間HLEC細胞活力的吸光度(A)值活力存在變化,第1、3、5天各實驗組間HLEC細胞活力的A值分彆是:空白對照組0.88、1.28、1.32；暘性對照組0.73、1.02、1.06；0.001％石決明提取液組0.73、1.03、1.06;0.01％石決明提取液組0.76、1.10、1.13;0.1％石決明提取液組0.79、1.22、1.21；0.3％石決明提取液組0.79、1.21、1.21；組間比較差異具有統計學意義(1d時F=23 922.42,p =0.05;3 d時,F=120 605.86,P＜0.05；5d時,F=150 939.45,P＜0.05).H2O2使細胞的活力降低,石決明提取液提高其活力,且在一定的時間和濃度範圍內具有依賴性,其中第3天時0.1％石決明提取液組HLEC細胞活力的A值為最高(1.22).H2O2組損傷後,HLEC中SOD、GSH的水平降低(SOD 158.05 U/mgprot、GSH 15.05 mg/1000 mgprot)、而MDA量增高到18.11 nmol/mgprot,石決明提取液組使氧化損傷所緻HLEC中抗氧化水平提高(SOD 188.64 U/mgprot、GSH 21.05 mg/ 1000 mgprot),脂質過氧化物升高有下降(MDA 14.16 nmol/mgprot),組間比較差異均具有統計學意義(P＜0.05).改變具有統計學意義(SOD:F=983.04,P＜0.05; GSH:F =444.44,P＜0.05; MDA:F=830.52,P＜0.05);暘性對照組中可見細胞覈染色質濃縮、聚集,石決明組中細胞染色質的聚集有不同程度的減輕.結論 石決明提取液對體外培養HLEC氧化損傷具有保護作用,可提高細胞內抗氧化水平,減少有害產物的生成.
목적 연구석결명제취액대양화손상적인정상체상피세포(HLEC)적작용.방법 실험연구.채용쌍양수(H2O2)간예체외배양HLEC양화손상모형,동시가입불동농도(0.001％、0.01％、0.1％、0.3％)적석결명제취액,분성공백대조조、양성대조조즉H2O2조화불동농도적석결명간예조,우1、3、5 d시용CCK-8검측각조HLEC적활력,도치상차현미경하관찰세포증식화형태개변,3d후용화학비색법검측초양화물기화매(SOD)、환원형곡광감태(GSH)화병이철(MDA)적수평.채용단인소방차분석진행양본균수간적다중비교,조간량량비교채용LSD-t검험.결과 불동시간점시각실험조간HLEC세포활력적흡광도(A)치활력존재변화,제1、3、5천각실험조간HLEC세포활력적A치분별시:공백대조조0.88、1.28、1.32；양성대조조0.73、1.02、1.06；0.001％석결명제취액조0.73、1.03、1.06;0.01％석결명제취액조0.76、1.10、1.13;0.1％석결명제취액조0.79、1.22、1.21；0.3％석결명제취액조0.79、1.21、1.21；조간비교차이구유통계학의의(1d시F=23 922.42,p =0.05;3 d시,F=120 605.86,P＜0.05；5d시,F=150 939.45,P＜0.05).H2O2사세포적활력강저,석결명제취액제고기활력,차재일정적시간화농도범위내구유의뢰성,기중제3천시0.1％석결명제취액조HLEC세포활력적A치위최고(1.22).H2O2조손상후,HLEC중SOD、GSH적수평강저(SOD 158.05 U/mgprot、GSH 15.05 mg/1000 mgprot)、이MDA량증고도18.11 nmol/mgprot,석결명제취액조사양화손상소치HLEC중항양화수평제고(SOD 188.64 U/mgprot、GSH 21.05 mg/ 1000 mgprot),지질과양화물승고유하강(MDA 14.16 nmol/mgprot),조간비교차이균구유통계학의의(P＜0.05).개변구유통계학의의(SOD:F=983.04,P＜0.05; GSH:F =444.44,P＜0.05; MDA:F=830.52,P＜0.05);양성대조조중가견세포핵염색질농축、취집,석결명조중세포염색질적취집유불동정도적감경.결론 석결명제취액대체외배양HLEC양화손상구유보호작용,가제고세포내항양화수평,감소유해산물적생성.
Objective To study the influence of haliotidis extractive on the oxidative damage in the human lens epithelial cells cultured in vitro.Methods Experimental study.Cultured human lens epithelial cells in vitro were intervened with hydrogen peroxide caused oxidative damage model,at the same time added different concentrations of concha haliotidis extractive.With control experiment research cells were divided into the blank control group,positive control group hydrogen peroxide group and hydrogen peroxide and different concentrations of concha haliotidis group,and on the first,third,fifth day the activity of Cultured human lens epithelial cells were detected with Cell Counting Kit-8 (CCK-8),cellular proliferation and morphological changes were observed with interred phase contrast microscope,and then on the third day chemical colorimetric were used to detect the homogenates superoxide dismutase(SOD),glutathione(GSH) and malondialdehyde(MDA)level.Results (1)At different time points there were variations between the activity of HLEC in each experimental group,Among each experimental group HLEC OD value of the cell vitality at 1 d,3 d,5 d,respectively were blank control group:0.88,1.28,1.32 ;Positive control group:0.73,1.02,1.06; 0.001％ concha haliotis extract group:0.73,1.03,1.06; 0.01％ concha haliotis extract group:0.76,1.10,1.13 ; 0.1％ concha haliotis extract group:0.79,1.22,1.21 ; 0.3％ concha haliotis extract group:0.79,1.21,1.21 ; the difference between groups was statistically significant (P ＜ 0.05) (1 d,F=23922.42,P＜0.05;3 d,F=120605.86,P＜0.05;5 d,F=150939.45,P＜0.05).H2O2 made the vitality of the cells reduce,concha haliotidis enhance its vitality,and in a certain range of time and concentrations there was dependence,with which the third day and 0.1％ was the best.(2)After adding H2O2,the SOD and GSH level of HLEC reduced,(SOD 158.05 U/mgprot,GSH 15.05 mg/gprot) but MDA increased to 18.11 nmol/mgprot,concha haliotidis groups made the increase of antioxidant level (SOD 188.64 U/mgprot,GSH 21.05 mg/1000 mgprot)and the decrease of lipid peroxidation in oxidative damaged HLECs(MDA 14.16 nmol/mgprot),change had a statistical significance (P ＜ 0.05) (SOD:F =983.04,P ＜0.05; GSH:F =444.44,P ＜0.05; MDA:F =830.52,P ＜0.05).(3) The chromatin of the positive control group concentrated and aggregated obviously,the aggregation of chromatin in coneha haliotidis group lightened.Conclusion The concha haliotidis can protect the cultured human lens epithelial cells in vitro which are oxidative injured,increased intracellular antioxidant levels,reduce the generation of hazardous products.
