本文選題：N-乙酰半胱氨酸 + 亞硒酸鈉��； 參考：《鄭州大學(xué)》2010年碩士論文

【摘要】： 研究目的 觀察N-乙酰半胱氨酸(N-acetylcysteine, NAC)對(duì)亞硒酸鈉誘導(dǎo)的大鼠白內(nèi)障的預(yù)防和治療作用,為白內(nèi)障的藥物防治提供實(shí)驗(yàn)依據(jù)。對(duì)象與方法 (一)預(yù)防作用實(shí)驗(yàn) 10天齡SD大鼠30只隨機(jī)分3組： 正常對(duì)照組-1(Al組)：每日腹腔注射0.1ml/10g體重劑量的生理鹽水,1次／日,共注射6天。 亞硒酸鈉組(B1組)：按3.46mg/kg(即20μmol/kg體重)頸部皮下注射亞硒酸鈉,隔日一次,共注射3次。注射亞硒酸鈉30分鐘內(nèi),按0.1ml/10g體重腹腔注射生理鹽水,共注射6天。 NAC預(yù)防組(C1組)：給10天齡SD大鼠按3.46mg/kg(即20μmol/kg體重)頸部皮下注射亞硒酸鈉,隔日一次,共注射3次。注射亞硒酸鈉30分鐘內(nèi),按0.1ml／10g體重的劑量腹腔注射濃度為2mmol/L的試劑NAC,1次／日。共注射6天。 (二)治療作用實(shí)驗(yàn) 白內(nèi)障模型制備：10天齡SD大鼠30只,按3.46mg/kg(即20μmol/kg體重)頸部皮下注射亞硒酸鈉,隔日一次,共注射3次。將造模成功的大鼠隨機(jī)分為病理對(duì)照組(15只)和NAC治療組(15只) 病理對(duì)照組(B2組)：每日腹腔注射0.1ml/10g體重的生理鹽水,共注射1月。 NAC治療組(C2組)：按0.1ml/10g體重的劑量腹腔注射濃度為2mmol/L的試劑NAC,1次／日。共注射1月。 正常對(duì)照組-2(A2組)：選預(yù)防實(shí)驗(yàn)中5只正常對(duì)照組大鼠,繼續(xù)按0.1ml／l0g體重的劑量腹腔注射生理鹽水,1次／日。共注射30天。 大鼠睜眼后裂隙燈下定期觀察大鼠晶狀體的渾濁情況并拍照；最后一次注藥后,分批處死大鼠,取出眼球,分離晶狀體。光鏡和掃描電鏡下觀察晶狀體上皮組織學(xué)和超微結(jié)構(gòu)的改變；免疫組化觀察亞硒酸鈉對(duì)晶狀體上皮細(xì)胞中caspase-3的影響。生化方法測(cè)定亞硒酸鈉對(duì)大鼠晶狀體組織中超氧化物歧化酶(SOD)、丙二醛(MDA)含量的影響。 結(jié)果 小劑量隔日頸部皮下注射20μmol/kg亞硒酸鈉溶液均可安全地誘導(dǎo)出大鼠白內(nèi)障動(dòng)物模型。 1.裂隙燈觀察：亞硒酸鈉組和病理對(duì)照組大鼠晶狀體混濁情況持續(xù)性加重,在注藥后第3天大部分晶狀體核出現(xiàn)環(huán)狀或塵埃狀混濁,七天均形成典型的核性白內(nèi)障,正常對(duì)照組晶狀體均保持透明,未見(jiàn)混濁；預(yù)防組晶狀體混濁程度低于亞硒酸鈉組,而高于正常對(duì)照組；治療組大鼠晶狀體混濁程度與相應(yīng)的病理對(duì)照組無(wú)明顯差異。 2.光鏡和掃描電鏡觀察：正常對(duì)照組晶狀體組織結(jié)構(gòu)基本正常,其他各組晶狀體超微組織結(jié)構(gòu)有不同程度的改變。其中預(yù)防實(shí)驗(yàn)中,亞硒酸鈉組損傷最重,NAC預(yù)防組次之,正常對(duì)照組最輕,三組差別顯著。治療試驗(yàn)中,病理對(duì)照組和NAC治療組無(wú)明顯差異。 3.免疫組化觀察：晶狀體上皮細(xì)胞caspase-3在正常對(duì)照組呈陰性表達(dá),平均光密度為250.319±2.347；NAC預(yù)防組呈弱陽(yáng)性表達(dá),平均光密度值為180.942±4.379,在亞硒酸鈉組呈強(qiáng)陽(yáng)性表達(dá),平均光密度值為70.312±14.216,三組比較差異均有有顯著性意義(P0.05)；在NAC治療組和病理對(duì)照組均呈強(qiáng)陽(yáng)性表達(dá),平均光密度值分別為100.467±4.477,71.437±11.329,兩組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),但兩組分別與正常對(duì)照組相比差異具有統(tǒng)計(jì)學(xué)意義(P0.05) 4.生化指標(biāo)分析：NAC預(yù)防組晶狀體中的SOD的活性高于亞硒酸鈉組(P0.05),而低于正常對(duì)照組(P0.05)；NAC治療組晶狀體中的SOD的活性明顯低于正常對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05),而與相應(yīng)的病理對(duì)照組比較無(wú)明顯差異(P0.05)。NAC預(yù)防組晶狀體中的MDA含量低于亞硒酸鈉組(P0.05),而高于正常對(duì)照組(P0.05)。NAC治療組晶狀體中的MDA含量與相應(yīng)的病理對(duì)照組比較無(wú)明顯差異(P0.05)。 結(jié)論 1.對(duì)SD大鼠皮下小劑量多次注射亞硒酸鈉可成功制備硒性白內(nèi)障模型。 2.亞硒酸鈉可引起LECs凋亡 3.NAC可以通過(guò)提高晶狀體組織中SOD的活性,減少M(fèi)DA生成,降低caspase-3的活性來(lái)減輕亞硒酸鈉引起的晶狀體損傷。 4.NAC對(duì)早期白內(nèi)障的發(fā)生、發(fā)展有一定的延緩和預(yù)防作用。 5.試劑NAC對(duì)于已經(jīng)形成的硒性白內(nèi)障無(wú)明顯的治療作用。
[Abstract]:Purpose of study



Objective To observe the preventive and therapeutic effects of N - acetyl cysteine ( NAC ) on sodium selenite - induced cataract in rats .



( 1 ) Preventive Effect Experiment



30 - day - old SD rats were randomly divided into 3 groups :



Normal control group - 1 ( Al group ) : 0.1 ml / 10 g body weight of physiological saline was injected intraperitoneally once daily for 6 days .



Sodium selenite group ( group B1 ) : The sodium selenite was subcutaneously injected at 3.46mg / kg ( that is , 20 渭mol / kg body weight ) in the neck of the neck , once daily for 3 times . After 30 minutes of sodium selenite injection , the saline was injected intraperitoneally with 0.1 ml / 10g body weight for 6 days .



NAC prevention group ( C1 group ) : 10 - day - old SD rats were given subcutaneous injection of sodium selenite at 3.46mg / kg ( that is , 20 渭mol / kg body weight ) , once daily for 3 times . After injection of sodium selenite for 30 minutes , a dose of 2 mmol / L of reagent NAC was injected intraperitoneally at a dose of 0.1 ml / 10 g body weight for 6 days .



( II ) Treatment effect experiment



The rats were randomly divided into pathological control group ( 15 rats ) and NAC treatment group ( 15 rats ) .



Pathological control group ( B2 group ) : 0.1 ml / 10g of normal saline was injected into the abdominal cavity every day for a total of 1 month .



NAC treatment group ( C2 group ) : A dose of 2 mmol / L reagent NAC was injected intraperitoneally at a dose of 0.1 ml / 10 g body weight for 1 time / day . A total of 1 month was injected .



Normal control group - 2 ( group A2 ) : 5 rats in normal control group were injected with normal saline at a dose of 0.1 ml / kg body weight for 30 days .



The opacity of the rat lens was observed and photographed under the slit lamp after open eyes .
After the last injection , the rats were sacrificed in batches , the eyeball was taken out , the lens was separated , and the changes of the epithelium and ultrastructure of the lens were observed under the light microscope and scanning electron microscope .
The effect of sodium selenite on caspase - 3 in lens epithelial cells was observed by immunohistochemistry . The effects of sodium selenite on superoxide dismutase ( SOD ) , malondialdehyde ( MDA ) content in lens tissue of rat lens were determined by biochemical method .



Results



The rat model of cataract was induced safely by subcutaneous injection of 20 渭mol / kg sodium selenite solution at low dose .



1 . Observation of slit lamp : The lens opacity in the sodium selenite group and the pathological control group was aggravated continuously . After the injection , most of the lens nuclei appeared cyclic or dust - like cloudiness , and typical nuclear cataract was formed on the seventh day , and the lens in the normal control group remained transparent , and the opacity was not seen ;
The degree of opacity in the prevention group was lower than that of the sodium selenite group , which was higher than that of the normal control group .
There was no significant difference between the degree of lens opacity and the corresponding pathological control group in the treatment group .



2 . Light microscope and scanning electron microscope observation : The structure of lens tissue of normal control group was basically normal , and the structure of the other groups had different degree of change . Among them , the injury of sodium selenite group was the most serious , NAC prevention group was the second , the normal control group was the lightest , and the three groups had significant difference . In the treatment trial , there was no significant difference between the pathological control group and NAC treatment group .



3 . Immunohistochemical observation showed that caspase - 3 was negative in normal control group , and the average optical density was 250.319 鹵 2.347 ;
The average optical density was 180.942 鹵 4.379 , and the mean optical density was 70.312 鹵 14.216 . There was significant difference between the three groups ( P0.05 ) .
The mean optical density was 100.467 鹵 4.477 , 71.437 鹵 11.329 , and the difference was not statistically significant ( P0.05 ) .



4 . Analysis of biochemical indexes : The activity of SOD in the lens of NAC prevention group was higher than that of sodium selenite group ( P0.05 ) , but lower than that of control group ( P0.05 ) .
The activity of SOD in the lens of NAC treatment group was significantly lower than that of the control group ( P0.05 ) . The content of MDA in the lens of NAC prevention group was lower than that of the normal control group ( P0.05 ) . The content of MDA in the lens of NAC treatment group was lower than that of the normal control group ( P0.05 ) .



Conclusion



1 . The selenium - induced cataract model was successfully prepared by multiple injections of sodium selenite into SD rats .



2 . Sodium selenite can cause apoptosis of LECs



3 . NAC can reduce the lens damage caused by sodium selenite by increasing the activity of SOD in lens tissue , reducing MDA formation and decreasing the activity of caspase - 3 .



4 . NAC has a certain delay and preventive effect on the occurrence and development of early cataract .



5 . NAC has no obvious therapeutic effect on the selenium - induced cataract already formed .
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R776.1

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