波动性高糖对人视网膜色素上皮细胞氧化应激的影响
파동성고당대인시망막색소상피세포양화응격적영향
The effect of intermittent high glucose on oxidative stress in human retinal pigment epithelial cells
  • 万方数据
    中国综合临床 중국종합림상
    CLINICAL MEDICINE OF CHINA
    2014年 2期 177-180 ,共4页

    李立琴%李志红%张云良%赵玮%郭淑芹%王翯%马文彬%刘莉芳%程晓东

    리립금%리지홍%장운량%조위%곽숙근%왕학%마문빈%류리방%정효동

    高血糖波动%人视网膜色素上皮细胞%氧化应激 高血糖波動%人視網膜色素上皮細胞%氧化應激
    고혈당파동%인시망막색소상피세포%양화응격
    Intermittent high glucose%Human retinal pigment epithelial cell%Oxidative stress
    目的 对比研究波动性高糖与恒定高糖对人视网膜色素上皮(HRPE)细胞氧化应激的影响.方法 培养HRPE细胞株,根据培养条件不同分为4组:(1)正常对照组:5.5 mmol/L葡萄糖;(2)恒定高糖组:33.0 mmol/L葡萄糖;(3)波动性高糖组:5.5 mmol/L和33.0 mmol/L葡萄糖波动组,日间每3小时高糖、2小时低糖交替3次,高糖过夜;(4)渗透压控制组:33.0 mmol/L甘露醇.每组均设6个复孔,置于37℃、5％ CO2孵箱中,分别于培养24、48、72 h收集细胞培养上清液用于过氧化物歧化酶(SOD)、谷胱甘肽(GSH)、丙二醛检测.结果 (1)培养24 h,波动性高糖组SOD为(12.1 ±3.0) U/ml,GSH为(68.5±3.8) mg/L,丙二醛为(17.5±3.0) μmol/L;恒定高糖组SOD为(21.8±1.6) U/ml,GSH为(90.8±4.8) mg/L,丙二醛为(12.9±1.2) μmol/L;正常对照组SOD为(31.1±4.7) U/ml,GSH为(143.4±8.3) mg/L,丙二醛为(3.1±1.1) μmol/L;渗透压控制组SOD为(32.4±2.8) U/ml,GSH为(143.3±12.8)mg/L,丙二醛为(2.8±1.2) μmol/L.(2)培养48 h,波动性高糖组SOD为(9.6±1.8) U/ml,GSH为(72.5±4.3)mg/L,丙二醛为(19.1 ±1.7) μmol/L;恒定高糖组SOD为(21.0±2.5) U/ml,GSH为(93.4±4.6) mg/L,丙二醛为(11.6±2.2)μmol/L;正常对照组SOD为(31.0±2.5) U/ml,GSH为(145.5±6.6) mg/L,丙二醛为(3.9±1.0) μmol/L;渗透压控制组SOD为(28.4±0.8) U/ml,GSH为(142.0 ±4.9)mg/L,丙二醛为(2.9±0.8)μmol/L.(3)培养72 h,波动性高糖组SOD为(10.9±1.7)U/ml,GSH为(70.9±7.3) mg/L,丙二醛为(19.5±0.5)μmol/L;恒定高糖组SOD为(20.4±1.7)U/ml,GSH为(91.6±7.2) mg/L,丙二醛为(11.2±3.3) μmol/L;正常对照组SOD为(32.4±4.4)U/ml,GSH为(143.0±23.2) mg/L,丙二醛为(3.0±1.0)μmol/L;渗透压控制组SOD为(29.5±2.6)U/ml,GSH为(134.5±11.1) mg/L,丙二醛为(2.8±0.8)μmol/L.(4)培养24、48、72 h波动性高糖组、恒定高糖组与正常对照组相比,差异均有统计学意义(P均＜0.01),波动性高糖组与恒定高糖组相比,差异均有统计学意义(P均＜0.01),渗透压控制组与正常对照组相比差异均无统计学意义(P均＞0.05),与恒定高糖组、波动性高糖组相比,差异均有统计学意义(P均＜0.01).结论 波动性高糖较恒定高糖对HRPE细胞有更强的氧化应激损伤.
    목적 대비연구파동성고당여항정고당대인시망막색소상피(HRPE)세포양화응격적영향.방법 배양HRPE세포주,근거배양조건불동분위4조:(1)정상대조조:5.5 mmol/L포도당;(2)항정고당조:33.0 mmol/L포도당;(3)파동성고당조:5.5 mmol/L화33.0 mmol/L포도당파동조,일간매3소시고당、2소시저당교체3차,고당과야;(4)삼투압공제조:33.0 mmol/L감로순.매조균설6개복공,치우37℃、5％ CO2부상중,분별우배양24、48、72 h수집세포배양상청액용우과양화물기화매(SOD)、곡광감태(GSH)、병이철검측.결과 (1)배양24 h,파동성고당조SOD위(12.1 ±3.0) U/ml,GSH위(68.5±3.8) mg/L,병이철위(17.5±3.0) μmol/L;항정고당조SOD위(21.8±1.6) U/ml,GSH위(90.8±4.8) mg/L,병이철위(12.9±1.2) μmol/L;정상대조조SOD위(31.1±4.7) U/ml,GSH위(143.4±8.3) mg/L,병이철위(3.1±1.1) μmol/L;삼투압공제조SOD위(32.4±2.8) U/ml,GSH위(143.3±12.8)mg/L,병이철위(2.8±1.2) μmol/L.(2)배양48 h,파동성고당조SOD위(9.6±1.8) U/ml,GSH위(72.5±4.3)mg/L,병이철위(19.1 ±1.7) μmol/L;항정고당조SOD위(21.0±2.5) U/ml,GSH위(93.4±4.6) mg/L,병이철위(11.6±2.2)μmol/L;정상대조조SOD위(31.0±2.5) U/ml,GSH위(145.5±6.6) mg/L,병이철위(3.9±1.0) μmol/L;삼투압공제조SOD위(28.4±0.8) U/ml,GSH위(142.0 ±4.9)mg/L,병이철위(2.9±0.8)μmol/L.(3)배양72 h,파동성고당조SOD위(10.9±1.7)U/ml,GSH위(70.9±7.3) mg/L,병이철위(19.5±0.5)μmol/L;항정고당조SOD위(20.4±1.7)U/ml,GSH위(91.6±7.2) mg/L,병이철위(11.2±3.3) μmol/L;정상대조조SOD위(32.4±4.4)U/ml,GSH위(143.0±23.2) mg/L,병이철위(3.0±1.0)μmol/L;삼투압공제조SOD위(29.5±2.6)U/ml,GSH위(134.5±11.1) mg/L,병이철위(2.8±0.8)μmol/L.(4)배양24、48、72 h파동성고당조、항정고당조여정상대조조상비,차이균유통계학의의(P균＜0.01),파동성고당조여항정고당조상비,차이균유통계학의의(P균＜0.01),삼투압공제조여정상대조조상비차이균무통계학의의(P균＞0.05),여항정고당조、파동성고당조상비,차이균유통계학의의(P균＜0.01).결론 파동성고당교항정고당대HRPE세포유경강적양화응격손상.
    Objective To compare the effects of intermittent and constant high glucose on oxidative stress and inflammatory injury in human retinal pigment epithelial cells (HRPE).Methods HRPE cells were caltured and randomly divided into 4 groups:normal glucose control group (5.5 mmol/L of glucose,NG group),consistent high glucose group (33.0 mmol/L of glucose,HG group),intermittent high glucose group (33.0 mmol/l of glucose for 3 hours,at an interval of two hours of 5.5 mmol/Lof glucose; Such intermittent exposure was repeated 3 times a day and the cells were exposed to 33.0 mmol/L of glucose in the night,IHG group) and high mannitol group (33.0 mmol/L of mannitol,MG group).Each group had 6 duplicate vials.Samples were cultured in incubator of 37℃,5％ CO2,and the activity of total superoxide (T-SOD) and the levels of malondialdehyde,and glutathione (GSH) in supernatant were assayed using chromatometry in 24 hours,48 hours,and 72 hours.Results (1) The activity of T-SOD,GSH and malonaldehyde(MDA) in the HG were (12.1 ±3.0) U/ml,(68.5 ±3.8) mg/L and (17.5 ±3.0) μmol/L respectively,in IHG groups were (21.8 ± 1.6) U/ml,(90.8 ±4.8) mg/L and (12.9 ± 1.2) μmol/L respectively,in control group were (31.1 ± ±4.7) U/ml,(143.4 ± 8.3) mg/L and (3.1 ± 1.1) μmol/L respectively,in MG group were (32.4 ± 2.8) U/ml,(143.3 ± 12.8) mg/L and (2.8 ± 1.2) μmol/L respectively.(2) When cells were cultured for 48 hours,the levels of T-SOD,GSH and MDA in the IHG,HG,NG,and MG groups were (9.6 ± 1.8) U/ml,(72.5 ±4.3) mg/L,(19.1 ±1.7) μmol/L;(21.0±2.5) U/ml,(93.4 ±4.6) mg/L,(11.6±2.2) μmol/L;(31.0±2.5) U/ml,(145.5±6.6) mg/L,(3.9±1.0) μmol/L;(28.4±0.8) U/ml,(142.0±4.9) mg/L,and (2.9 ± 0.8) μmol/L respectively.(3) When cells were cultured for 72 hours,the levels of T-SOD,GSH and MDA in the IHG,HG,NG,and MG groups were (10.9 ± 1.7) U/ml,(70.9 ±7.3) mg/L,(19.5 ±0.5)μmol/L;(20.4± 1.7) U/ml,(91.6±7.2) mg/L,(11.2±3.3) μmol/L;(32.4±4.4) U/ml,(143.0 ± ±23.2) mg/L,(3.0±1.0) μmol/L;(29.5 ±2.6) U/ml,(134.5 ±11.1) mg/L,and (2.8 ±0.8) μmol/L respectively.(4) Compared with control group,the activity of T-SOD and the levels of GSH and MDA in IHG,HG group were significantly different after culture for 24 h,48 h,and 72 h (P ＜ 0.01).There was significant difference between the IHG group and the HG group in terms of the activity of T-SOD and the levels of GSH and MDA (P ＜ 0.01).The activity of T-SOD and the levels of GSH and MDA of the MG group were significant different from IHG and the HG groups (P ＜0.01).But no significant difference was seen in the NG group (P ＞ 0.05).Conclusion Intermittent high glucose causes a more significant damage in terms of oxidate stress and inflammatory injury to HRPE cells than that of consistent high glucose.
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