氧化和抗氧化指標(biāo)在大鼠高脂飲食所致脂肪肝中的動(dòng)態(tài)變化

1、氧化和抗氧化指標(biāo)在大鼠高脂飲食所致脂肪肝中的動(dòng)態(tài)變化     【摘要】  目的 探討氧應(yīng)激和脂質(zhì)過氧化在高脂飲食所致脂肪性肝炎形成中的作用。方法 雄性SD大鼠 30只隨機(jī)分為3組，每組10只。正常組普通飼料喂養(yǎng)；模型、組喂高脂飼料；12周末處死正常組、模型組大鼠；模型組大鼠繼續(xù)喂養(yǎng)至16周末處死，測定血清轉(zhuǎn)氨酶（ALT、AST），全血谷胱甘肽過氧化物酶（GSHPX），肝勻漿丙二醛（MDA）含量，超氧化物歧化酶（SOD）活性，總抗氧化能力（TAOC），一氧化氮（NO），誘導(dǎo)型一氧化氮合酶（iNOS），谷胱甘肽（GSH）水平，觀察肝組織學(xué)改變。結(jié)果

2、 模型組大鼠12周出現(xiàn)單純性脂肪肝，16周發(fā)展為脂肪性肝炎。與正常組比，模型組大鼠肝組織脂質(zhì)過氧化產(chǎn)物MDA含量明顯增多，而抗氧化物SOD，GSH，GSHPX含量明顯降低，且肝臟的脂肪變性嚴(yán)重程度隨著高脂飲食喂養(yǎng)的時(shí)間延長而加劇。結(jié)論 氧應(yīng)激和脂質(zhì)過氧化在肝組織脂肪變性后炎癥的過程中發(fā)揮重要作用。 【關(guān)鍵詞】  氧應(yīng)激脂質(zhì)過氧化非酒精性脂肪性肝炎Development Changes of Oxidative and Antioxidative Indexes in Establishment of Steatohepatitis in Rats Fed by Highfat Die

3、tsPeng Haiying, Yu Hongbo, Zuo Zhong (Dalian Sanatorium of Shenyang Military Region, Dalian 116013, China; PLA No.210 Hospital, Dalian 116021, China)Abstract： Objective  To explore the role of oxygen stress and lipid peroxidation in the establishment of steatohepatitis in rats fed by highf

4、at diets. Methods  Thirty male Sprague Dawley rats were assigned into three groups (n=10) at random. The rats fed by normal foods served as controls. The rats in model groups (, ) were fed with high-fat diets. The animals in control group and model I group were killed at the end of the 12th wee

5、k, and the ones left were killed at the end of the 16th week. Blood samples were collected for the detection of serum aspartate transaminase (AST), alanine aminotransferase (ALT) and glutathione peroxidase (GSHPX), and liver tissues were obtained for the detection of malondialdehyde (MDA) contents,

6、superoxide dismutase (SOD) activity, total antioxidative capacity (TAOC), and nitric oxide (NO), inducible nitric oxide synthase (iNOS) and glutathione (GSH) levels. The histological changes were observed under light microscope. Results  Pure fatty liver was formed in the model rats after in th

7、e 12th week, and fatty hepatitis was established in the 16th week. In comparison with those of normal rats, the contents of MDA were increased (P<0.01) but antioxide SOD, GSH, GSHPX contents were decreased significantly (P<0.05, or P<0.01). The severity of hepatic fatty degeneration aggrava

8、ted with the prolonging of the highfat diets taking duration.  Conclusion  Oxidative stress and lipid peroxidation may play an important role in the formation of liver inflammation cell infiltration.Key  words： oxygen stress; lipid peroxidation; nonalcoholic steatohepatitis研究認(rèn)為氧應(yīng)激和脂質(zhì)過

9、氧化在非酒精性脂肪性肝炎(nonalcoholic  steatohepatitis，NASH)發(fā)病中占有重要地位13。我們通過高脂飲食建立NASH大鼠模型，動(dòng)態(tài)檢測氧化和抗氧化指標(biāo)觀察實(shí)驗(yàn)性NASH肝組織的氧化損傷。1  材料和方法1.1  材料  雄性SD大鼠 30只，體重（150±10）g，購自大連醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心。膽固醇購自上海伯奧生物工程公司，豬油自備。超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)、誘導(dǎo)型一氧化氮合酶(iNOS)、總抗氧化能力(TAOC)、谷胱甘肽過氧化物酶(GSHPX)、還原型谷胱甘肽 (GSH)

10、等試劑盒購自南京建成生物工程公司。1.2  方法  大鼠普通飼料喂養(yǎng)1周后，隨機(jī)分為3組(每組10只)。正常組普通飼料喂養(yǎng)；模型、組喂高脂飼料(880 g/kg普通飼料+100 g/kg 豬油+20 g/kg 膽固醇)。各組大鼠自由進(jìn)食、進(jìn)水，分籠(每籠5只)飼養(yǎng)于（20±2）、明暗各12 h 的動(dòng)物實(shí)驗(yàn)室內(nèi)。12周末處死正常組、模型組大鼠；模型組大鼠繼續(xù)喂養(yǎng)至16周末處死。方法為禁食12 h，乙醚麻醉，下腔靜脈采血后處死，迅速取出肝臟，按常規(guī)制備血清、肝勻漿、肝組織石蠟切片標(biāo)本，肝組織于-70 低溫保存。（1）采用全自動(dòng)生化分析儀測定血清ALT、AST，全血GS

11、HPX (DTNB直接法)；（2）肝勻漿測定MDA含量(改進(jìn)后的硫代巴比妥酸熒光法)，SOD活性 (改良的鹽酸羥胺法)，TAOC、NO、iNOS、GSH(按試劑盒說明操作)；（3）肝臟病理學(xué)檢查，光鏡下評估脂肪變性程度和炎癥活動(dòng)度計(jì)分45。1.3  統(tǒng)計(jì)學(xué)處理  計(jì)量資料以均值±標(biāo)準(zhǔn)差（±s）表示，各組間實(shí)驗(yàn)數(shù)據(jù)采用兩樣本均數(shù)的t檢驗(yàn)；等級資料采用秩和檢驗(yàn)。2  結(jié)果2.1  血清ALT、AST及全血GSHPX變化  與正常組比較，模型，組ALT，AST顯著增高(P0.01)，GSHPX顯著下降(P0.01)，詳見表1。表1  各組大鼠血清生化指標(biāo)（略）與正常組比較，P0.012.2  肝MDA、NO、iNOS、SOD、TAOC及GSH變化  與正常組比較，模型組MDA顯著增高，SOD、GSH下降明顯(P0.05)；模型組MDA、iNOS、NO顯著增高(P0.01，P0.05)；模型組SOD、TAOC、GSH增高(P0.01)，詳見表2。表2  各組大鼠肝勻漿生化指標(biāo)（略）與正