龙胆苦苷对非酒精性脂肪肝病大鼠脂肪酸过氧化酶体β氧化信号通路的影响研究
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摘要
目的观察龙胆苦苷对非酒精性脂肪肝病(NAFLD)大鼠的作用,并从过氧化酶体β氧化途径探究其作用机制。方法 72只雄性清洁级SD大鼠在适应性喂养10 d后按照随机数字法分为正常组、模型组、多烯磷脂酰胆碱组及龙胆苦苷低、中、高剂量组,每组12只。正常组大鼠给予普通饲料饲养,其余5组大鼠给予高脂饲料和5%红糖水饮用饲养,12周后检验造模成功后,正常组、模型组予500μL/(kg·d)生理盐水灌胃,多烯磷脂酰胆碱组予多烯磷脂酰胆碱23 mg/(kg·d)灌胃,龙胆苦苷低、中和高剂量组分别予50,100,200 mg/(kg·d)龙胆苦苷灌胃,时间均为8周。称体质量,处死大鼠测定肝质量,计算肝质量占体质量百分比;取血检测肝功能[谷草转氨酶(AST)和谷丙转氨酶(ALT)]、血脂[胆固醇(TC)和三酰甘油(TG)];HE染色进行肝脏病理学观察,Western blot和RT-PCR法检测肝脏组织中乙酰辅酶A氧化酶(ACOX1)和过氧化物酶体增殖物激活受体α(PPARα)蛋白及mRNA表达情况。结果模型组大鼠表现为明显的多饮多食、肥胖和精神倦怠,龙胆苦苷各剂量组上述症状明显较模型组好转。模型组大鼠肝脏切片出现明显脂肪空泡样改变,龙胆苦苷各剂量组脂肪空泡数量明显较模型组减少。模型组大鼠肝质量、体质量、肝质量占体质量百分比及血清AST、ALT、TC、TG水平均明显高于正常组(P均<0.05),多烯磷脂酰胆碱组和龙胆苦苷各剂量组除体质量与模型组相近外,其余指标均明显低于模型组(P均<0.05),且龙胆苦苷高剂量组血清AST、ALT、TC、TG水平均明显低于多烯磷脂酰胆碱组(P均<0.05)。模型组ACOX1和PPARα蛋白及mRNA表达量均明显低于正常组(P均<0.05),多烯磷脂酰胆碱组和龙胆苦苷各剂量组ACOX1和PPARα蛋白及mRNA表达量均明显高于模型组(P均<0.05),且龙胆苦苷各剂量组ACOX1和PPARα蛋白表达量及龙胆苦苷高剂量组ACOX1和PPARαmRNA表达量均明显高于多烯磷脂酰胆碱组(P均<0.05)。结论龙胆苦苷可明显改善NAFLD大鼠症状及肝功能,降低血脂,可能与其可以调控肝脏过氧化酶体β氧化关键酶ACOX1和PPARα的表达有关。
Objective It is to observe the effect of gentiopicroside(GP) on rats with non-alcoholic fatty liver disease(NAFLD) and explore its mechanism of action from the pathogenesis of peroxidase β oxidation. Methods Seventy-two male clean SD rats were randomly divided into normal group, model group, polyene phosphatidylcholine(PPC) group and low, medium and high dose of GP groups after 10 days of adaptive feeding, 12 rats in each group. The rats in the normal group were fed with normal diet, the ones in the other groups were fed with high fat diet plus 5% brown sugar water for 12 weeks to establish NAFLD rat models. After the models were successfully established, the rats in normal group and model group were given 500 μL/(kg·d) normal saline by gavage, the ones were given 23 mg/(kg·d) PPC in PPC group and 50, 100, 200 mg/(kg·d) GP in the low, medium and high dose groups respectively by gavage. All the rats were treatment for 8 weeks. The body mass was measured, the rats were sacrificed to measure liver mass, and the percentage of liver mass/body mass was calculated; the blood was taken to detect liver function(AST, and ALT), blood lipids(TC,TG); HE staining for liver pathology, Western blot and RT-PCR for detection of acetyl-CoA oxidase(ACOX1) and peroxisome proliferator-activated receptor alpha(PPARα) protein and mRNA expression in liver tissue. Results The rats in the model group showed obvious polydipsia, obesity and mental fatigue, and the above symptoms of every GP group were significantly better than those of the model group. The liver slices of the model group showed obvious vacuolar changes, and the number of fat vacuoles in the every GP group was significantly lower than that of the model group. The liver mass, body mass, percentage of liver mass/body mass and the levels of serum AST, ALT, TC, TG in the model group were significantly higher than those in the normal group(P<0.05), all the indexes above except body mass in PPC group and every GP group were lower than those in the model group(P<0.05), and the serum levels of AST, ALT, TC and TG in the high dose group of GP were significantly lower than that in the PPC group(P<0.05). The expressions of ACOX1 and PPARα protein and mRNA in the model group were significantly lower than those in the normal group(P<0.05). The expressions of ACOX1 and PPARα protein and mRNA in the PPC group and every GP group were significantly higher than those in the model group(P<0.05), and the expression of ACOX1 and PPARα in every GP group and the expression of ACOX1 and PPARα mRNA in the high dose group were significantly higher than those in the PPC group(P<0.05). Conclusion GP can significantly improve the symptoms and liver function and reduce blood lipids of rats with NAFLD, its mechanism may be related to the regulation of the expression of the key enzymes of hepatic peroxidase β oxidation ACOX1 and PPARα.
Objective It is to observe the effect of gentiopicroside(GP) on rats with non-alcoholic fatty liver disease(NAFLD) and explore its mechanism of action from the pathogenesis of peroxidase β oxidation. Methods Seventy-two male clean SD rats were randomly divided into normal group, model group, polyene phosphatidylcholine(PPC) group and low, medium and high dose of GP groups after 10 days of adaptive feeding, 12 rats in each group. The rats in the normal group were fed with normal diet, the ones in the other groups were fed with high fat diet plus 5% brown sugar water for 12 weeks to establish NAFLD rat models. After the models were successfully established, the rats in normal group and model group were given 500 μL/(kg·d) normal saline by gavage, the ones were given 23 mg/(kg·d) PPC in PPC group and 50, 100, 200 mg/(kg·d) GP in the low, medium and high dose groups respectively by gavage. All the rats were treatment for 8 weeks. The body mass was measured, the rats were sacrificed to measure liver mass, and the percentage of liver mass/body mass was calculated; the blood was taken to detect liver function(AST, and ALT), blood lipids(TC,TG); HE staining for liver pathology, Western blot and RT-PCR for detection of acetyl-CoA oxidase(ACOX1) and peroxisome proliferator-activated receptor alpha(PPARα) protein and mRNA expression in liver tissue. Results The rats in the model group showed obvious polydipsia, obesity and mental fatigue, and the above symptoms of every GP group were significantly better than those of the model group. The liver slices of the model group showed obvious vacuolar changes, and the number of fat vacuoles in the every GP group was significantly lower than that of the model group. The liver mass, body mass, percentage of liver mass/body mass and the levels of serum AST, ALT, TC, TG in the model group were significantly higher than those in the normal group(P<0.05), all the indexes above except body mass in PPC group and every GP group were lower than those in the model group(P<0.05), and the serum levels of AST, ALT, TC and TG in the high dose group of GP were significantly lower than that in the PPC group(P<0.05). The expressions of ACOX1 and PPARα protein and mRNA in the model group were significantly lower than those in the normal group(P<0.05). The expressions of ACOX1 and PPARα protein and mRNA in the PPC group and every GP group were significantly higher than those in the model group(P<0.05), and the expression of ACOX1 and PPARα in every GP group and the expression of ACOX1 and PPARα mRNA in the high dose group were significantly higher than those in the PPC group(P<0.05). Conclusion GP can significantly improve the symptoms and liver function and reduce blood lipids of rats with NAFLD, its mechanism may be related to the regulation of the expression of the key enzymes of hepatic peroxidase β oxidation ACOX1 and PPARα.
引文
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[15] 聂青和,王全楚.过氧化酶体β氧化与脂肪性肝炎[J].肝脏,2001,6(3):190-192
[16] 王昆,杨茂,陈昱江.PPAR在中医药治疗NAFLD中的影响[J/CD].世界最新医学信息文摘:电子版,2015,15(77):36-37
[2] 范建高.非酒精性脂肪性肝病进展——非酒精性脂肪性肝病的流行率[J].中国医师进修杂志,2006,29(9A):1-2
[3] 马振增,陆伦根.非酒精性脂肪性肝病与肝硬化[J].实用肝脏病杂志,2016,19(2):135-138
[4] 梁乾飞,赵彩彦.非酒精性脂肪性肝病与肝细胞癌发生[J].实用肝脏病杂志,2016,19(1):113-116
[5] 姜少灏,康丽娟,蒋晔,等.龙胆及其复方中龙胆苦苷在大鼠体内的药动学研究[J].中国药学杂志,2005,40(3):212-215
[6] 武俊紫,贾亚敏,沈平瑞,等.富硒灵芝调控大鼠乙酰辅酶A羧化酶α表达治疗非酒精性脂肪肝病的研究[J].天然产物研究与开发,2014,26(7):1086-1092
[7] 武俊紫.艾塞那肽对非酒精性脂肪肝病的治疗作用研究[D].昆明:昆明理工大学,2014
[8] 张晓兰,张志钢.非酒精性脂肪性肝病治疗进展[J].中国医师进修杂志,2010,33(4):6-9
[9] 黄婷婷,周玉荣,张向君,等.非酒精性脂肪肝治疗进展[J].医学综述,2015,21(8):1406-1409
[10] 李军祥,王允亮.非酒精性脂肪性肝病中西医结合研究存在的问题思考[C].中华中医药学会脾胃病分会全国脾胃病学术交流会,2015
[11] 杜启雪,王仁忠.龙胆泻肝汤治疗肝火上炎型突发性耳聋临床研究[J].亚太传统医药,2015,11(9):93-94
[12] 李艳秋,赵德化.龙胆苦甙抗鼠肝损伤的作用[J].医学争鸣,2001,22(18):1645-1649
[13] 刘占文,陈长勋,金若敏,等.龙胆苦苷的保肝作用研究[J].中草药,2002,33(1):47-50
[14] 殷杰,李万平.非酒精性脂肪肝的发病机制研究进展[J].四川生理科学杂志,2012,34(3):126-130
[15] 聂青和,王全楚.过氧化酶体β氧化与脂肪性肝炎[J].肝脏,2001,6(3):190-192
[16] 王昆,杨茂,陈昱江.PPAR在中医药治疗NAFLD中的影响[J/CD].世界最新医学信息文摘:电子版,2015,15(77):36-37
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