中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
1期
70-74
,共5页
王明玲%胡淑芳%曹建雄%王安安%肖天林
王明玲%鬍淑芳%曹建雄%王安安%肖天林
왕명령%호숙방%조건웅%왕안안%초천림
糖尿病眼部并发症/角膜、晶状体、视网膜%醛脱氢酶%脂质过氧化
糖尿病眼部併髮癥/角膜、晶狀體、視網膜%醛脫氫酶%脂質過氧化
당뇨병안부병발증/각막、정상체、시망막%철탈경매%지질과양화
Diabetic ocular complication/cornea,lens,retina%Aldehyde dehydrogenases%Lipid peroxidation
背景 糖尿病并发症的发生与脂质过氧化有关,而醛脱氢酶(ALDH)是已知的抗脂质过氧化物之一,研究ALDH在糖尿病眼组织中的活性与表达量的变化能为糖尿病性眼部并发症的诊断和治疗提供依据. 目的 研究ALDH在不同时期糖尿病大鼠眼组织中的活性与质量分数的变化趋势,探讨其在糖尿病所致眼部疾病发病过程中的作用机制.方法 将健康7～8周龄清洁级雄性SD大鼠28只按随机数字表法分为糖尿病模型组和正常对照组.糖尿病模型组大鼠腹腔注射65 mg/kg溶于柠檬酸钠缓冲液的链脲佐菌素(STZ),血糖达16.7 ～ 22.2 mmol/L为造模成功.正常对照组大鼠腹腔以同样的方式注射等量柠檬酸钠缓冲液.于造模成功后的2个月、4个月分别处死7只大鼠,制备大鼠角膜、晶状体及视网膜匀浆液,测定蛋白质质量浓度.采用M5多功能酶标仪测量角膜、晶状体及视网膜各组织中ALDH活性,采用酶联免疫吸附(ELISA)法检测ALDH在角膜、晶状体和视网膜匀浆中的表达量. 结果 模型建立后2个月、4个月糖尿病模型组大鼠血糖水平明显高于正常对照组,而体质量明显低于正常对照组,差异均有统计学意义(均P=0.000).造模后2个月、4个月时糖尿病模型组大鼠角膜、晶状体及视网膜中ALDH活性均明显高于相应时间点的正常对照组大鼠,差异均有统计学意义(F=396.601,P=0.000)；上述各组织中ALDH活性(A 340值)均随着造模时间的延长而升高,差异有统计学意义(F=53.139,P=0.000),其中4个月时糖尿病模型组大鼠角膜、晶状体、视网膜中ALDH活性较2个月时高,差异均有统计学意义(P＜0.01),而正常对照组大鼠角膜、晶状体及视网膜中的ALDH活性在2个时间点间相比差异均无统计学意义(P＞0.05)；2个时间点正常对照组与糖尿病模型组大鼠眼组织中ALDH活性均为角膜最高,视网膜次之,晶状体最低,差异有统计学意义(F=6973.000,P=0.000).造模后2个月、4个月时糖尿病模型组大鼠角膜、晶状体及视网膜匀浆中ALDH表达量均明显高于正常对照组大鼠,差异有统计学意义(F=312.985,P=0.000)；2个组大鼠ALDH表达均随着造模时间的延长而增加,差异有统计学意义(F=19.203,P=0.000),2个时间点各组大鼠眼组织中ALDH表达量均表现为角膜最高,视网膜次之,晶状体最低,差异有统计学意义(F=3243.000,P=0.000). 结论 ALDH可减轻糖尿病大鼠眼组织的脂质氧化损伤,可能在预防糖尿病相关性眼部并发症发生方面发挥一定的作用.
揹景 糖尿病併髮癥的髮生與脂質過氧化有關,而醛脫氫酶(ALDH)是已知的抗脂質過氧化物之一,研究ALDH在糖尿病眼組織中的活性與錶達量的變化能為糖尿病性眼部併髮癥的診斷和治療提供依據. 目的 研究ALDH在不同時期糖尿病大鼠眼組織中的活性與質量分數的變化趨勢,探討其在糖尿病所緻眼部疾病髮病過程中的作用機製.方法 將健康7～8週齡清潔級雄性SD大鼠28隻按隨機數字錶法分為糖尿病模型組和正常對照組.糖尿病模型組大鼠腹腔註射65 mg/kg溶于檸檬痠鈉緩遲液的鏈脲佐菌素(STZ),血糖達16.7 ～ 22.2 mmol/L為造模成功.正常對照組大鼠腹腔以同樣的方式註射等量檸檬痠鈉緩遲液.于造模成功後的2箇月、4箇月分彆處死7隻大鼠,製備大鼠角膜、晶狀體及視網膜勻漿液,測定蛋白質質量濃度.採用M5多功能酶標儀測量角膜、晶狀體及視網膜各組織中ALDH活性,採用酶聯免疫吸附(ELISA)法檢測ALDH在角膜、晶狀體和視網膜勻漿中的錶達量. 結果 模型建立後2箇月、4箇月糖尿病模型組大鼠血糖水平明顯高于正常對照組,而體質量明顯低于正常對照組,差異均有統計學意義(均P=0.000).造模後2箇月、4箇月時糖尿病模型組大鼠角膜、晶狀體及視網膜中ALDH活性均明顯高于相應時間點的正常對照組大鼠,差異均有統計學意義(F=396.601,P=0.000)；上述各組織中ALDH活性(A 340值)均隨著造模時間的延長而升高,差異有統計學意義(F=53.139,P=0.000),其中4箇月時糖尿病模型組大鼠角膜、晶狀體、視網膜中ALDH活性較2箇月時高,差異均有統計學意義(P＜0.01),而正常對照組大鼠角膜、晶狀體及視網膜中的ALDH活性在2箇時間點間相比差異均無統計學意義(P＞0.05)；2箇時間點正常對照組與糖尿病模型組大鼠眼組織中ALDH活性均為角膜最高,視網膜次之,晶狀體最低,差異有統計學意義(F=6973.000,P=0.000).造模後2箇月、4箇月時糖尿病模型組大鼠角膜、晶狀體及視網膜勻漿中ALDH錶達量均明顯高于正常對照組大鼠,差異有統計學意義(F=312.985,P=0.000)；2箇組大鼠ALDH錶達均隨著造模時間的延長而增加,差異有統計學意義(F=19.203,P=0.000),2箇時間點各組大鼠眼組織中ALDH錶達量均錶現為角膜最高,視網膜次之,晶狀體最低,差異有統計學意義(F=3243.000,P=0.000). 結論 ALDH可減輕糖尿病大鼠眼組織的脂質氧化損傷,可能在預防糖尿病相關性眼部併髮癥髮生方麵髮揮一定的作用.
배경 당뇨병병발증적발생여지질과양화유관,이철탈경매(ALDH)시이지적항지질과양화물지일,연구ALDH재당뇨병안조직중적활성여표체량적변화능위당뇨병성안부병발증적진단화치료제공의거. 목적 연구ALDH재불동시기당뇨병대서안조직중적활성여질량분수적변화추세,탐토기재당뇨병소치안부질병발병과정중적작용궤제.방법 장건강7～8주령청길급웅성SD대서28지안수궤수자표법분위당뇨병모형조화정상대조조.당뇨병모형조대서복강주사65 mg/kg용우저몽산납완충액적련뇨좌균소(STZ),혈당체16.7 ～ 22.2 mmol/L위조모성공.정상대조조대서복강이동양적방식주사등량저몽산납완충액.우조모성공후적2개월、4개월분별처사7지대서,제비대서각막、정상체급시망막균장액,측정단백질질량농도.채용M5다공능매표의측량각막、정상체급시망막각조직중ALDH활성,채용매련면역흡부(ELISA)법검측ALDH재각막、정상체화시망막균장중적표체량. 결과 모형건립후2개월、4개월당뇨병모형조대서혈당수평명현고우정상대조조,이체질량명현저우정상대조조,차이균유통계학의의(균P=0.000).조모후2개월、4개월시당뇨병모형조대서각막、정상체급시망막중ALDH활성균명현고우상응시간점적정상대조조대서,차이균유통계학의의(F=396.601,P=0.000)；상술각조직중ALDH활성(A 340치)균수착조모시간적연장이승고,차이유통계학의의(F=53.139,P=0.000),기중4개월시당뇨병모형조대서각막、정상체、시망막중ALDH활성교2개월시고,차이균유통계학의의(P＜0.01),이정상대조조대서각막、정상체급시망막중적ALDH활성재2개시간점간상비차이균무통계학의의(P＞0.05)；2개시간점정상대조조여당뇨병모형조대서안조직중ALDH활성균위각막최고,시망막차지,정상체최저,차이유통계학의의(F=6973.000,P=0.000).조모후2개월、4개월시당뇨병모형조대서각막、정상체급시망막균장중ALDH표체량균명현고우정상대조조대서,차이유통계학의의(F=312.985,P=0.000)；2개조대서ALDH표체균수착조모시간적연장이증가,차이유통계학의의(F=19.203,P=0.000),2개시간점각조대서안조직중ALDH표체량균표현위각막최고,시망막차지,정상체최저,차이유통계학의의(F=3243.000,P=0.000). 결론 ALDH가감경당뇨병대서안조직적지질양화손상,가능재예방당뇨병상관성안부병발증발생방면발휘일정적작용.
Background Diabetic complication is associated with lipid peroxidation.Aldehyde dehydrogenases (ALDH) catalyze the irreversible oxidation of a variety of biological aldehydes,including lipid-derived aldehydes (LDAs),and thus protect organs and tissues from toxic LDAs.Understanding the activity of ALDH in different ocular tissues in diabetic subjects is very important for prevention and treatment of diabetic ocular complications.Objective This research aimed to investigate the activity and expression of ALDH in different ocular tissues in diabetic rats and to explore the mechanism of ALDH in diabetes-induced eye disease.Methods Twenty-eight healthy SPF male Sprague-Dawley(SD) rats weighted 170-180 g were randomly divided into the normal control group and diabetic group.The diabetic animal model was established by intraperitonial injection of 4％ streptozotocin at 65 mg/kg.Isometric citric acid buffer was injected in the rats of the normal control group.The rats were sacrificed in each group 2 and 4 months after the establishment of the diabetic models,and eyeballs were obtained for the preparation of corneal,lens and retinal homogenates.ALDH activity was detected using a multifunctional microplate reader SpectraMax M5,and ALDH content was measured by ELISA at the wavelength of 450 nm with the SpectraMax M5 ELISA reader.Results The blood glucose level in diabetic rats was significantly elevated at various time points compared with the normal control group(P=0.000),and body weights were evidently lower in the diabetic group than in the normal group (P =0.000).The activities of ALDH (A340) in corneal,lens and retinal tissues in the diabetic group were increased in comparison with the normal control group (F =396.601,P=0.000),and showed an enhancement with the lapsing of time (F =53.139,P =0.000).In addition,the highest level of ALDH was found in the cornea and the lowest level in the lens(F =6973.000,P=0.000).The expression level of ALDH in the corneal,lens and retinal homogenates was significantly higher in the diabetic group compared with the normal control group (F=312.985,P =0.000) and showed a considerable increase over the course (F =19.203,P=0.000).The highest expression level was seen in the cornea and the lowest was in the lens,with a significant difference among these three kinds of tissues (F =3243.000,P =0.000).Conclusions ALDH can protect ocular tissue from the damage of lipid peroxidation.Thess results suggest that ALDH plays a role in preventing diabetes-related ocular complications.
