蒜氨酸通过CBS/CSE途径改善小鼠非酒精性脂肪肝病的研究

doi:10.16431/j.cnki.1671-7236.2023.12.042

摘要/Abstract

摘要： 【目的】研究蒜氨酸对非酒精性脂肪肝病(NAFLD)小鼠肝脏中胱硫醚-β-裂解酶(cystathionine-β-synthase,CBS)、胱硫醚-γ-裂解酶(cystathionine-γ-lyase,CSE) 表达的影响,阐释蒜氨酸改善NAFLD的作用途径,为临床药物研发提供科学依据。【方法】50只雄性C57/6J小鼠随机分为5组:正常组、模型组、硫氢化钠组、蒜氨酸组和蒜氨酸＋PAG组(DL-炔丙基甘氨酸,DL-propargylglycine,PAG),每组10只。正常组饲喂普通饲料,其他4组饲喂高脂饲料。蒜氨酸组灌胃给予蒜氨酸20 mg/kg;蒜氨酸＋PAG组灌胃给予蒜氨酸20 mg/kg,同时腹腔注射PAG 15 mg/kg;硫氢化钠组腹腔注射硫氢化钠2 mg/kg;正常组和模型组灌胃给予等剂量的蒸馏水,每天1次,每周称体重1次,预防给药6周后处死,观察肝脏形态学和病理学变化,检测血清中总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白(high density lipoprotein,HDL-C)、低密度脂蛋白(low density lipoprotein,LDL-C)的水平,Western blotting检测肝脏组织中 CBS和CSE蛋白表达水平。【结果】与正常组相比,模型组小鼠体重显著增加(P＜0.05),其他组小鼠体重无显著变化(P＞0.05)。形态学观察显示,模型组小鼠肝脏偏黄无光泽,成花斑状,其他组小鼠肝脏表面光滑、质地均匀、颜色与正常组小鼠接近。肝脏病理学HE染色可见,正常组切片正常,模型组出现明显脂肪空泡和脂滴沉积,说明脂肪肝模型建立成功;与模型组相比,硫氢化钠组和蒜氨酸组均有明显好转;与蒜氨酸组相比,蒜氨酸＋PAG组抗脂肪变性作用减弱。与正常组相比,模型组小鼠血清中TG、TC、LDL-C均显著升高(P＜0.05),各给药组小鼠TC、TG、LDL-C相较于模型组显著降低(P＜0.05)。与正常组相比,模型组小鼠CBS和CSE蛋白表达水平显著下降(P＜0.05),硫氢化钠组和蒜氨酸组干预后CBS和CSE蛋白表达水平显著或极显著上调(P＜0.05;P＜0.01);与模型组相相比,蒜氨酸＋PAG组小鼠CBS和CSE表达无显著差异(P＞0.05)。【结论】蒜氨酸可明显增加高脂饮食诱导的NAFLD小鼠肝脏组织中CBS和CSE表达,改善血清中血脂各项指标,减少小鼠肝细胞脂质沉积,发挥保护肝脏作用。

关键词: 蒜氨酸; 小鼠; 非酒精性脂肪肝病(NAFLD); 胱硫醚-β-裂解酶(CBS); 胱硫醚-γ-裂解酶(CSE)

Abstract: 【Objective】 The experiment was aimed to investigate the effect of alliin on the expression of cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) in liver of mice with nonalcoholic fatty liver disease (NAFLD),in order to elucidate the pathway of action of alliin to improve NAFLD and provide scientific basis for clinical drug development.【Method】 Fifty male C57/6J mice were randomly divided into five groups:Normal group,model group,sodium hydrosulfide group,allicin group and allicin＋PAG group (DL-propargylglycine,PAG),with 10 mice in each group.The normal group was fed with normal feed and the other four groups were fed with high fat feed.The allicin group was given 20 mg/kg allicin by gavage.Allicin＋PAG group was given 20 mg/kg of allicin by gavage,and at the same time,15 mg/kg PAG was injected intraperitoneally.Sodium hydrosulfide group was injected with 2 mg/kg sodium hydrosulfide by intraperitoneal injection.Normal and model groups were given equal doses of distilled water by gavage,once a day,weighed once a week.After 6 weeks of prophylactic administration,mice were euthanized to observe liver morphology and pathological changes,and the serum total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL-C),and low density lipoprotein (LDL-C) were detected.The protein expression of CBS and CSE in liver of mice were detected by Western blotting.【Result】 Compared with normal group,the body weight of mice in model group was increased significantly (P＜0.05),there was no significant change in the body weight of mice in other groups (P＞0.05).Morphological observation results showed that the liver of mice in model group were yellowish,lusterless and flowery,while the liver of mice in the other groups had a smooth surface,uniform texture and a color close to that of mice in normal group.HE staining of liver pathology showed that the slices of the normal group were normal,while the model group showed obvious fat vacuoles and lipid droplet deposition,indicating that the fatty liver model was successfully established.Compared with model group,there was a significant improvement in sodium hydrosulfide group and the allicin group.Antilipidemic effect of allicin＋PAG group was weakened compared with allicin group.TG,TC and LDL-C in serum of mice were significantly higher in model group compared with normal group (P＜0.05),and TC and TG were significantly lower in all administered groups compared with model group (P＜0.05).Compared with normal group,the protein expression of CBS and CSE of mice were significantly decreased in model group (P＜0.05),and the protein expression of CBS and CSE in mice were significantly or extremely up-regulated in sodium hydrosulfide and allicin groups after the intervention (P＜0.05 or P＜0.01).There was no significant difference in the expression of CBS and CSE in the allicin ＋ PAG group compared with model group (P＞0.05).【Conclusion】 Allicin could significantly increase the expression of CBS and CSE in liver of NAFLD mice induced by high-fat diet,improve the indicators of lipids in serum,reduce the lipid deposition in mouse hepatocytes,and play a role in protecting the liver.

Key words: alliin; mice; nonalcoholic fatty liver disease (NAFLD); cystathionine-β-synthase (CBS); cystathionine-γ-lyase (CSE)

中图分类号:

S859.7

马红梅. 蒜氨酸通过CBS/CSE途径改善小鼠非酒精性脂肪肝病的研究[J]. 中国畜牧兽医, 2023, 50(12): 5213-5221.

MA Hongmei. Study on Alliin Improving Nonalcoholic Fatty Liver Disease in Mice Through CBS/CSE Pathway[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5213-5221.

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