三种非酒精性脂肪肝炎模型的比较
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上海中医药大学附属曙光医院肝病研究所

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中药药源性肝损伤的真实世界临床研究


Comparison of three Non-alcohol Steatohepatitis Models
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Institute of Liver Disease,Shuguang Hospital, affiliated to Shanghai University of Traditional Chinese Medicine

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    摘要:

    目的:比较四氯化碳注射复合高脂饲料饮食(CCl4+HFD)、蛋氨酸胆碱缺乏饮食(MCD)以及高脂高糖高胆固醇饮食(AMLN)诱导的三种NASH模型血清学及病理学特征,并初步探讨不同NASH模型的病理机制。方法:分别采用四氯化碳注射复合高脂饲料饮食(CCl4+HFD)、蛋氨酸胆碱缺乏饮食(MCD)、高脂高糖高胆固醇饮食(AMLN)喂养10周、8周、26周制备小鼠NASH模型。比色法检测血清ALT、AST、GLU的水平及肝组织TG、TC、SOD、MDA含量;ELISA法检测血清FINS含量并计算HOMA-IR指数; HE、天狼猩红及油红O染色观察肝组织炎症、肝细胞脂肪变及胶原沉积,并根据活动度评分(NAS)对肝组织脂肪性肝炎分级评分。结果:与对照小鼠相比,三种模型小鼠血清ALT、AST活性、肝脏TG、TC、SOD、MDA含量均有显著上升;其中AMLN饮食诱导及CCl4+HFD复合模型小鼠的血清FINS、GLU的含量显著升高, HOMA-IR指数显著升高; MCD模型小鼠血清FINS的含量及HOMA-IR指数显著下降。HE染色、油红O染色及NAS评分结果显示,3种模型小鼠的肝组织均已进展到脂肪性肝炎阶段,且CCl4+HFD模型组小鼠肝组织胶原沉积最明显,AMLN模型小鼠肝脏脂滴最丰富。结论:上述三种模型均可稳定模拟人类NASH疾病的血清学及病理学变化,其中AMLN模型可模拟人类的发病过程及机制,且胰岛素抵抗、氧化应激等全身代谢紊乱表现,但耗时较长,纤维化进展较慢。MCD饮食8周即可模拟NASH的血清及病理学特征,但无肥胖胰岛素抵抗发生。CCl4复合HFD模型10周即可诱导NASH模型,能模拟其血清学及病理学变化,且肝组织纤维沉积与氧化应激损伤明显。

    Abstract:

    Objective Compare the serological and pathological characteristics of three nonalcoholic steatohepatitis (NASH) models: high fat diet (HFD) with carbon tetrachloride (CCl4) injection, methionine and choline deficient diet (MCD) and Aymlin liver NASH (AMLN) diet induced NASH models, and make primary study on their pathological mechanisms. Methods Three NASH models were established by feeding HFD with CCl4 injection for 10 weeks, MCD for 8 weeks and Amylin diet for 26 weeks. After feeding, serum transaminase (ALT, AST) and glucose (GLU), liver triglyceride (TG), total cholesterol (TC), Superoxide dismutase (SOD) and Malondialdehyde (MDA) levels were measured. Insulin level was measured by ELISA and HOMA-IR index was calculated. Hematoxylin-eosin (HE) staining, Sirius red staining and Oil Red staining were used to indicate the pathological changes of liver. Furthermore, NAS score was used to evaluate the pathology grade. Results Compared to normal control mice, all mice in three model groups had obvious increase in serum transaminase, liver TG, TC, SOD, MDA level. The level of serum FINS, GLU and HOMA-IR index were significantly increased in mice of Amylin and HFD with CCl4 group, but decreased in mice of MCD group. According to HE staining, Oil Red staining and NAS score, all mice in three groups MCD mice had NASH phenotype changes. Liver collagen deposition was most obvious in mice of HFD with CCl4 group. Meanwhile, liver lipid droplets were most abundant in mice of AMLN group. Conclusions All three NASH models show different advantages and disadvantages. NASH models induced by Amylin diet recaptures the key NASH features observed in patients.

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  • 收稿日期:2024-01-17
  • 最后修改日期:2024-09-06
  • 录用日期:2024-12-02
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