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目的 研究表皮生长因子 (EGF)、神经降压素 (NT)、降钙素基因相关肽 (CGRP)对四氯化碳 (CCl4)急性损伤的在体大鼠肝脏和离体培养肝细胞的保护作用 ,并探讨其机制。 方法 第一步建立大鼠 CCl4损伤模型 ,设立对照组、损伤组及各胃肠肽保护组 ,于注射 CCl4前 30 min、前 10 min,后 10 min注射各胃肠肽 ,2 4h后测定血清酶学水平、肝匀浆 SOD活性及 MDA含量 ,观察肝脏形态学变化 ;第二步建立大鼠离体培养肝细胞损伤模型 ,设对照组、损伤组及不同浓度胃肠肽预处理组 (胃肠肽提前 1h加入 ) ,2 4h后测上清液酶学水平、肝细胞内 SOD活性和 MDA含量 ,以台盼蓝染色试验计算肝细胞存活率 ,观察形态学变化 ;第三步以 Fura- 2 / AM和 DPH荧光探针测定肝细胞内游离钙和膜流动性 ,观察对肝细胞有直接保护作用的胃肠肽对这两个指标的影响。 结果 (1) EGF、NT、CGRP明显降低 CCl4损伤后大鼠血清酶学水平、肝匀浆 MDA含量 ,使 SOD活性回升 (P<0 .0 1) ,改善肝脏病理学变化。(2 ) EGF、NT明显降低原代培养肝细胞 CCl4损伤后上清液酶学水平、MDA含量 ,提高 SOD活性和细胞存活率 (P<0 .0 1) ,改善形态学变化。CGRP对上述指标和形态学改变无明显影响。(3) EGF、NT明显对抗肝细胞 CCl4损伤后胞内 [Ca2 + ]i的增高和膜流动?
Objective To study the protective effects of epidermal growth factor (EGF), neurotensin (NT) and calcitonin gene related peptide (CGRP) on rat liver and in vitro cultured hepatocytes injured by carbon tetrachloride (CCl4) Role, and explore its mechanism. Methods The CCl4 injury model was established in rats, and the control group, injury group and protective group of gastrointestinal peptide were established. The gastrointestinal peptide was injected 30 minutes before injection of CCl4, 10 minutes before injection and 10 minutes after injection. Serum Enzyme activity, liver homogenate activity of SOD and MDA content were observed, and morphological changes of liver were observed. The second step was to establish rat hepatocyte injury model in vitro, and the control group, injury group and different concentrations of gastrointestinal peptide pretreatment group Enteric peptide 1h ahead of time). After 24 hours, the supernatant enzyme level, SOD activity in hepatocytes and MDA content were measured. The survival rate of hepatocytes was calculated by Trypan blue staining and the morphological changes were observed. The third step was Fura- 2 / AM and DPH fluorescence probe to measure the intracellular free calcium and membrane fluidity, to observe the direct protective effect of hepatocyte on the two indicators of gastrointestinal peptide. Results (1) EGF, NT and CGRP significantly decreased the levels of serum enzymes and the content of MDA in liver homogenate after CCl4 injury, and increased the activity of SOD (P <0.01), and improved the pathological changes of liver. (2) EGF, NT significantly reduced the level of serum enzyme, MDA content, SOD activity and cell survival rate (P <0.01) of supernatant after CCl4 injury in primary culture hepatocytes, and improved morphological changes. CGRP had no significant effect on the above indexes and morphological changes. (3) EGF and NT obviously antagonize the increase of intracellular [Ca2 +] i and membrane flow after CCl4 injury in hepatocytes.