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熱鹽水致大鼠萎縮性胃炎血清和胃黏膜組織SOD和TAC的變化
作者：江梅，張瀝，海春旭，秦緒軍，陶梅，張玲霞，曹廣州【關(guān)鍵詞】 萎縮性胃炎
Alterations of SOD and TAC in serum and gastric mucosa of atrophic gastritis rat caused by highsalt hot water
【Abstract】 AIM: To investigate the dynamic variation of superoxide dismutase (SOD) and total antioxidant capacity (TAC) in the process of atrophic gastritis formation caused by over salty hot diet. METHODS: The atrophic gastritis rat model was made by feeding 150 g/L salty water of 55℃ for 12 weeks. The normal control was fed with pure water of 25℃. The rats were killed group by group every 4 weeks and the serum and gastric mucosa were kept to test SOD and TAC. Tissue slice of gastric mucosa was observed under optic microscope. The SOD was tested by improved muriatic acid hydroxylamine method, the total albumen content of stomach homogenate was tested by Lowrys method and TAC was tested by TAC kits. RESULTS: In the 12th week, the gastric mucosa in rats fed with highsalt hot water presented with typical pathological change as atrophic gastritis, but this pathological change was absent in control group. SOD and TAC in serum and gastric mucosa had a perfect timeeffect relation in hotsalt water group. With the time prolonged, the SOD and TAC decreased significantly in hotsalt water group. CONCLUSION: Active oxygen damnification plays an important precursory role in the formation and evolution of atrophic gastritis.
【Keywords】 atrophic gastritis; highsalt hot diet; SOD; TAC
【摘要】 目的: 觀察過(guò)熱、過(guò)咸飲食引起胃黏膜損傷以致造成胃黏膜萎縮的過(guò)程中超氧化物歧化酶（SOD）及總抗氧化力（TAC）的動(dòng)態(tài)變化. 方法: 采用55℃，150 g/L NaCl連續(xù)灌胃12 wk，制成大鼠萎縮性胃炎模型，對(duì)照組采用等量的25℃水灌胃,在此期間每隔4 wk處死一批大鼠. 將大鼠麻醉后腹主動(dòng)脈取血分離血清，同時(shí)摘取胃組織，制成組織勻漿，分別測(cè)定血清及胃黏膜組織中SOD和TAC含量. SOD測(cè)定采用改良的鹽酸羥胺法， TAC采用南京建成生物公司的試劑盒，總蛋白測(cè)定采用Lowrys法. 結(jié)果：與對(duì)照組相比較，經(jīng)熱鹽水灌飼大鼠的血清和胃黏膜組織中SOD活性及TAC水平顯著降低（P＜0.05，P＜0.05），并且隨處理時(shí)間延長(zhǎng)，SOD活性和TAC降低愈顯著. 結(jié)論：SOD和TAC的顯著改變說(shuō)明活性氧損傷在萎縮性胃炎發(fā)生發(fā)展過(guò)程中起著重要的先導(dǎo)作用，也提示有效的抗氧化干預(yù)很可能對(duì)于預(yù)防萎縮性胃炎具有很好的防治作用.
【關(guān)鍵詞】 萎縮性胃炎；熱鹽水；超氧化物歧化酶；總抗氧化力
0引言
慢性萎縮性胃炎是臨床常見的胃部疾病，由于其與胃癌尤其是腸型胃癌的發(fā)生呈顯著正相關(guān)，1978年世界衛(wèi)生組織將其定為胃癌前狀態(tài)［1］. 我們以人類飲食中不可缺少的兩大因素食物的熱度與咸度，創(chuàng)建了快速建立慢性萎縮性胃炎動(dòng)物模型的方法，并觀察從正常胃黏膜發(fā)展到萎縮性胃炎的演變過(guò)程中超氧化物歧化酶（superoxide dismutase,SOD）和總抗氧化力（total antioxidant capacity,TAC）的動(dòng)態(tài)變化，以探討活性氧損傷與萎縮性胃炎發(fā)生的關(guān)系.
1材料和方法
1.1材料
健康、性成熟的雄性SD大鼠44只7 wk齡，體質(zhì)量200~250 g，由第四軍醫(yī)大學(xué)動(dòng)物實(shí)驗(yàn)中心提供. 鹽酸羥胺、Triton X100及NBT購(gòu)自華美生物公司，福林酚購(gòu)自鼎國(guó)生物公司，其余試劑均為國(guó)產(chǎn)分析純. TAC試劑盒購(gòu)自南京建成生物研究所. 722型光柵分光光度計(jì)為上海分析儀器總廠產(chǎn)品，SHZ881臺(tái)式水浴恒溫震蕩器為江蘇太倉(cāng)鹿河生化儀器廠產(chǎn)品. 采用55℃, 150 g/L NaCl連續(xù)灌胃12 wk，制成大鼠萎縮性胃炎模型［2］，對(duì)照組采用等量25℃水灌胃12 wk.
1.2方法
實(shí)驗(yàn)開始隨機(jī)宰殺４只作為空白“0 wk”對(duì)照. 其余大鼠分成對(duì)照組（control）20只、熱鹽水組（CAG）20只. 分別于4，8，12，24和32 wk各隨機(jī)�。粗唬�10 g/L戊巴比妥鈉溶液0.5 mL/kg ip麻醉，腹主動(dòng)脈取血分離血清，同時(shí)摘取全胃組織，制成組織勻漿，分別測(cè)定血清和組織中SOD, TAC和總蛋白的含量. SOD測(cè)定采用改良的鹽酸羥胺法，樣品管取樣品20 μL，非酶管用三蒸水代替，依次加入75 mmol/L，pH=10.2碳酸鹽緩沖液2.0 mL，3 mL/L Triton X100 0.3 mL，1.2 mmol/L 鹽酸羥胺0.5 mL，0.98 mmol/L NBT 0.1 mL，37℃水浴20 min，2.0 mL甲酸終止反應(yīng)，560 nm比色. 總蛋白測(cè)定采用Lowrys法. TAC按照試劑盒中的步驟嚴(yán)格操作. 另取胃竇黏膜組織，從胃大彎至胃小彎連續(xù)切片，組織切片進(jìn)行HE染色. 參照1994年美國(guó)休斯頓胃炎診斷分類標(biāo)準(zhǔn)及井岡山會(huì)議胃炎診斷分類標(biāo)準(zhǔn)對(duì)胃竇、胃體黏膜組織學(xué)各項(xiàng)指標(biāo)進(jìn)行評(píng)定，組織切片由專人采取盲法閱片.
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