红曲黄素对高脂血症小鼠血脂及肝脏AMPKα ACC、PPAR-α、CPT1蛋白表达的影响
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摘要
目的:研究红曲黄素(Ankaflavin,AK)对高脂血症小鼠的血脂调节及肝脏保护作用。方法:选用70只清洁级健康雄性C57BL/6J小鼠,适应性饲养1周,随机选取10只小鼠作为正常组,喂以基础饲料,其余小鼠喂以高脂饲料,连续6周。将成模小鼠随机分为5组,分别为模型组、AK低、中、高[50、100、200 mg/(kg·d)]剂量组和SIM[15 mg/(kg·d)]组,模型组与正常组灌胃给予等体积生理盐水,每组10只,每天灌胃1次,连续4周。末次给药后,各组小鼠麻醉后腹主动脉采血进行血清总胆固醇(TC)、三酰甘油(TG)、瘦素(Leptin)、谷草转氨酶(AST)及谷丙转氨酶(ALT)的含量测定。解剖取肝脏,测定肝脏组织中TC、TG含量,并通过HE染色观察肝脏组织形态变化、油红O染色观察肝脏脂质沉积情况;采用Western blot法检测小鼠肝脏中PPAR-α和CPT1的蛋白表达水平以及AMPKα和ACC的磷酰化情况。结果:与正常组相比,模型组小鼠的体质量、血清和肝组织TC、TG含量均显著升高(P<0.01或P<0.05);肝组织P-AMPKα、P-ACC、PPAR-α及CPT1蛋白的表达显著降低(P<0.01);肝细胞形成脂质沉积,胞浆内多出现脂肪空泡。AK干预后,小鼠体质量降低,血清中TC、TG、Leptin含量降低,AST、ALT含量降低及肝脏P-AMPKα、P-ACC、PPAR-α及CPT1蛋白表达水平显著增加,肝脏的脂肪性病变明显改善。结论:AK对高脂血症小鼠具有明显的降脂效果并有效减轻其肝脏损伤;其作用机制可能与激活AMPK途径刺激脂肪酸氧化密切相关。
Objective: To observe the effects of Ankaflavin(AK) on regulating blood lipid and alleviating liver impairment in hyperlipidemia mice. Methods:70 healthy male c57 bl/6 J mice of clean grade were used for experiment. Adaptive feeding for 1 week. 10 mice were randomly selected as normal group and fed with basic diet. While the rest of the mice were fed with a high-fat diet. Continuous modeling for 6 weeks. The model mice were randomly divided into five groups: the model group. low, middle and high doses(50,100,200 mg/kg·d~(-1))of AK.The model group and the normal group were given the same volume of normal saline, 10 rats in each group, once a day for 4 weeks. At the end of the experimental period, abdominal aorta after anesthesia for the determination of serum lipid parameters(including the content of serum TC,TG,Leptin,AST and ALT). Livers was separated from the euthanized mice. Determined the levels of TC and TG in liver. Morphological changes of livers and the lipid deposition in liver were observed using HE stain and oil red O stain respectively. Protein expression levels of PPAR-α,CPT1,P-AMPKα/AMPKα,P-ACC/ACC were detected by Western blot. Results: Compared with the normal group, body weight, serum lipid parameters and levels of TC and TG in liver were significantly higher in the model group(P < 0.01 or P <0.05). Protein expression levels of PPAR-α,CPT1,P-AMPKα/AMPKα,P-ACC/ACC were significantly decreased(P <0.01). Lipid deposition occurred in liver cells and much fat vacuoles occurred in the cytoplasm. After AK intervention. Mice weight loss, serum TC, TG, leptin content decreased, AST and ALT content decreased, liver P-AMPKα,P-ACC,PPAR-α and CPT1 protein expression levels increased significantly. Fatty lesions in the liver were significantly improved. Conclusion: The effects of AK on regulating blood lipid and alleviating liver impairment in hyperlipidemia mice were obviously, the mechanism of hypolipidemia and liver protection may be closely related to the activation of AMPK pathway to stimulate fatty acid oxidation.
Objective: To observe the effects of Ankaflavin(AK) on regulating blood lipid and alleviating liver impairment in hyperlipidemia mice. Methods:70 healthy male c57 bl/6 J mice of clean grade were used for experiment. Adaptive feeding for 1 week. 10 mice were randomly selected as normal group and fed with basic diet. While the rest of the mice were fed with a high-fat diet. Continuous modeling for 6 weeks. The model mice were randomly divided into five groups: the model group. low, middle and high doses(50,100,200 mg/kg·d~(-1))of AK.The model group and the normal group were given the same volume of normal saline, 10 rats in each group, once a day for 4 weeks. At the end of the experimental period, abdominal aorta after anesthesia for the determination of serum lipid parameters(including the content of serum TC,TG,Leptin,AST and ALT). Livers was separated from the euthanized mice. Determined the levels of TC and TG in liver. Morphological changes of livers and the lipid deposition in liver were observed using HE stain and oil red O stain respectively. Protein expression levels of PPAR-α,CPT1,P-AMPKα/AMPKα,P-ACC/ACC were detected by Western blot. Results: Compared with the normal group, body weight, serum lipid parameters and levels of TC and TG in liver were significantly higher in the model group(P < 0.01 or P <0.05). Protein expression levels of PPAR-α,CPT1,P-AMPKα/AMPKα,P-ACC/ACC were significantly decreased(P <0.01). Lipid deposition occurred in liver cells and much fat vacuoles occurred in the cytoplasm. After AK intervention. Mice weight loss, serum TC, TG, leptin content decreased, AST and ALT content decreased, liver P-AMPKα,P-ACC,PPAR-α and CPT1 protein expression levels increased significantly. Fatty lesions in the liver were significantly improved. Conclusion: The effects of AK on regulating blood lipid and alleviating liver impairment in hyperlipidemia mice were obviously, the mechanism of hypolipidemia and liver protection may be closely related to the activation of AMPK pathway to stimulate fatty acid oxidation.
引文
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[2] Chen J,Deng J,Zhang Y Y,et al.Study on regulatory effect of Danshensu on lipid metabolism of hyperlipidemia rats [J].China J Chin Mater Med (中国中药杂志),2015,40(2):313-317.
[3] Li X R,Chen J J,Liu X G.Lipid lowering activity of total flavonoids from Mulberry Leaves in two hyperlipidemia animal models [J].Chin Pharm J (中国药学杂志),2009,44(21):1630-1633.
[4] Alcala A,Jansen S,Tellez T,et al.Statins improve visual field alterations related to hypercholesterolemia [J].Athrosclerosis,2010,209(2):510-514.
[5] Yu S.Studies on the effect of Cassia Aqueous Extract and Aurantio-obtusin on hyperlipidemia rats based on metabolomics[D].Hu’nan:Hunan Normal University,2016.
[6] Lee C L,Wen J Y,Hsu Y W,et al.Monascus-fermented yellow pigments monascin and ankaflavin showed antiobesity effect via the suppression of differentiation and lipogenesis in obese rats fed a high-fat diet [J].J Agric Food Chem,2013,61(7):1493-1500.
[7] Hsu L C,Liang Y H,Hsu Y W,et al.Anti-inflammatory Properties of Yellow and Orange Pigments from Monascus purpureus NTU 568 [J].J Agric Food Chem,2013,61(11):2796-2802.
[8] Lee B H,Hsu W H,Pan T M.Red Mold Rice against hepatic inflammatory damage in Zn-deficient rats [J].J Trad Complem Med,2012,2(1):52-60.
[9] Hsu W H,Liao TH,Lee B H,et al.Ankaflavin regulates adipocyte function and attenuates hyperglycemia caused by high-fat diet via PPAR-γ activation [J].J Funct Foods,2013,5(1):124-132.
[10] Hu H M,Zhu Y C,Zhu Q Q,et al.Analysis on animal models of experimental hyperlipidemia [J].China J Chin Mater Med (中国中药杂志),2016,41(20):3709-3714.
[11] Gao Z Q.Influence of Baicalin on hyperlipemia and NF-κB signaling pathways induced by lipopolysaccha-ride.[D].Guangzhou:Guangzhou University of Chinese Medicine,2011.
[12] Guo Y R,Choung S Y.Germacrone attenuates hyperlipidemia and improves lipid metabolism in high-fat diet-induced obese C57BL/6J mice [J].J Med Food,2017,20(1):46-55.
[13] HU Tianjiao,JIANG Zhen,Zhang Wenyou,LI Jiping,LIU Moxiang.Effect of Water Extract of Radix Isatidis on Early Liver Injury in Diabetes Mellitus Rats[J].Chin J Mod Appl Pharm(中国现代应用药学),2017,34(2):196-199.
[14] LI Yingying,ZHANG Yin.Research progress on the pro-apoptotic mechanims of AMP-activated Protein Kinase in Islet Beta Cells[J].Chin J Mod Appl Pharm(中国现代应用药学),2018,35(1):143-147.
[15] Yamauchi T,Kamon J,Minokoshi Ya,et al.Adiponectin stimulates glucose and fatty-acid oxidation by activating AMP-activated protein kinase[J].Nat Med,2002,8:1288-1295.
[16] Hardie D.AMPK:A key regulator of energy balance in the single cell and the whole organism[J].Int J Obesity,2008,32:S7-S12.
[17] Moller D,Berger J.Role of PPARs in the regulation of obesity related insulin sensitivity and inflammation[J].Int J Obesity,2003,27:S17-S21.
[18] Villarroya F,Iglesias R,Giralt M.PPARs in the control of uncoupling proteins gene expression[J].PPAR Res,2007,2007:74364.