熊果酸改善自然衰老大鼠糖脂代谢紊乱与自噬能力的研究
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摘要
为研究熊果酸对自然衰老大鼠糖脂代谢紊乱和自噬功能影响。实验取22月龄SD雄性大鼠25只,随机分为3组,即老年模型组(n=9),熊果酸低剂量组(UA-L):灌胃给予熊果酸10 mg/kg(n=8),熊果酸高剂量组(UA-H):灌胃给予熊果酸50 mg/kg(n=8);另取8只3月龄SD青年大鼠作为青年对照组,均给予常规正常饮食饮水并记录摄食量和体重变化。用酶法和ELISA法检测血浆葡萄糖、甘油三酯和胰岛素并计算HOMA-IR指数,测定肝脏甘油三酯;油红"O"染色观察肝脏脂质沉积情况;Western blot法检测肝脏自噬相关蛋白及线粒体质量相关蛋白的表达。结果显示熊果酸(50 mg/kg)并不影响大鼠食物摄入量、体质量,但是逆转了老年大鼠血浆甘油三酯、胰岛素和HOMA-IR的升高,降低了肝质量、肝脏TG含量、改善了肝脏脂质沉积(P<0.05),与此同时老年大鼠肝脏自噬相关蛋白LC3B/3A,Beclin1和线粒体质量控制蛋白PGC-1α,Drp1的表达显著升高,但对P62、Pink1的表达无影响。根据以上实验结果推测熊果酸可以提高胰岛素敏感性,改善老年大鼠脂肪肝,增强肝细胞自噬,提高线粒体质量。
This study aims to investigate the effect of ursolic acid(UA) on glycolipid metabolism disorder and hepatic autophagy in naturally ageing rats.Twenty-five male aging rats were divided randomly into three groups:the natural aging rat group(Aging,n=9),the aging rats supplemented with ursolic acid at 10 mg/kg(UA-L,n=8) and 50 mg/kg(UA-H,n=8) for 7 weeks.Eight young rats were randomly selected as the young normal group(Young,n=8).Enzymatical method and ELISA were used for the examination of plasma glucose,triglycerides,insulin concentrations and HOMA-IR index was calculated;Liver triglycerides were measured by enzymatical method;Liver lipid deposition was observed by Oil Red "O" staining;The autophagy-related proteins and mitochondrial quality proteins were analysed by Western blot.Results showed that treatment with UA(50 mg/kg,once daily,by oral gavage) could attenuate age-associated high plasma TG and insulin concentration under fasting conditions as well as suppress the increase in the HOMA-IR index,which suggests there is an improvement of systemic insulin sensitivity.Moreover,liver weight and hepatic triglyceride content were also decreased.Attenuation of the increased Oil Red O staining area was evident on histological examination of liver in UA(50 mg/kg)-treated rats.Importantly,UA(50 mg/kg) administration reversed the decreased expressions of autophagy-related proteins LC3 B/3 A,Beclin1 and mitochondrial quality proteins PGC-1α,Drp1 in aging rats,although P62,Pink1 remained unchanged.The results suggest that UA may attenuate systematic insulin resistance and hepatic lipid deposition,enhance hepatic autophagy,improve mitochondrial quality in aging rats.
This study aims to investigate the effect of ursolic acid(UA) on glycolipid metabolism disorder and hepatic autophagy in naturally ageing rats.Twenty-five male aging rats were divided randomly into three groups:the natural aging rat group(Aging,n=9),the aging rats supplemented with ursolic acid at 10 mg/kg(UA-L,n=8) and 50 mg/kg(UA-H,n=8) for 7 weeks.Eight young rats were randomly selected as the young normal group(Young,n=8).Enzymatical method and ELISA were used for the examination of plasma glucose,triglycerides,insulin concentrations and HOMA-IR index was calculated;Liver triglycerides were measured by enzymatical method;Liver lipid deposition was observed by Oil Red "O" staining;The autophagy-related proteins and mitochondrial quality proteins were analysed by Western blot.Results showed that treatment with UA(50 mg/kg,once daily,by oral gavage) could attenuate age-associated high plasma TG and insulin concentration under fasting conditions as well as suppress the increase in the HOMA-IR index,which suggests there is an improvement of systemic insulin sensitivity.Moreover,liver weight and hepatic triglyceride content were also decreased.Attenuation of the increased Oil Red O staining area was evident on histological examination of liver in UA(50 mg/kg)-treated rats.Importantly,UA(50 mg/kg) administration reversed the decreased expressions of autophagy-related proteins LC3 B/3 A,Beclin1 and mitochondrial quality proteins PGC-1α,Drp1 in aging rats,although P62,Pink1 remained unchanged.The results suggest that UA may attenuate systematic insulin resistance and hepatic lipid deposition,enhance hepatic autophagy,improve mitochondrial quality in aging rats.
引文
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14 Sahni S,Merlot AM,et al.Gene of the month:BECN1[J].J Clin Pathol,2014,67:656-660.
15 Duran A,Amanchy R,Linares JF,et al.P62 is a key regulator of nutrient sensing in the mTORC1 pathway[J].Molecular Cell,2011,44:134-146.
16 Zhu YS,Liu JL.Cell autophagy and mitochondrial quality control with healthy aging[J].Prog Biochem Biophys (生物化学与生物物理进展),2014,41:247-256.
17 Fige MT,Osiewacz HD,Reichert AS.Quality control of mitochondria during aging:is there a good and a badside of mitochondrial dynamics[J].Bioessays,2013,35:314-322.
18 Yang CC,Chen D,Lee SS,et al.The dynamin-related protein DRP-1 and the insulin signaling pathway cooperate to modulate Caenorhabditis elegans longevity[J].Aging Cell,2011,10:724-728.
19 Vives-Bauza C,Zhou C,Huang Y,et al.PINK1-dependent recruitment of Parkin to mitochondria in mitophagy[J].Proc Natl Acad Sci USA,2010,107:378-383.
2 You TH,Wen L,Liu F.Recent advances on active substances of anti-aging and its mechanisms[J].Nat Prod Res Dev(天然产物研究与开发),2015,27:1985-1990.
3 Mao YQ,Fan XM.Autophagy,a new target for non-alcoholic fatty liver disease therapy[J].WCJD (世界华人消化杂志),2015,23:5107-5112.
4 Christian P,Sacco J,Adeli K.Autophagy:emerging roles in lipid homeostasis and metabolic control[J].Biochim Biophys Acta,2013,1831:819-824.
5 Mellor KM,Bell JR,Ritchie RH,et al.Myocardial insulin resistance,metabolic stress and autophagy in diabetes[J].Clin Exp Pharmacol Physiol,2013,40(1):56-61.
6 Jian L,Gao MJ,Xia X.Ursolic acid attenuate myocardial ischemia reperfusion injury in diabetic rats[J].Nat Prod Res Dev(天然产物研究与开发),2017,29:843-848.
7 Marisa A,Cagri C,Carla M Sa,et al.Ursolic acid and luteolin-7-glucoside improve lipid profiles and increase liver glycogen content through glycogen synthase kinase-3[J].Phytother Res,2010,24:220-224.
8 Bahrami SA,Bakhtiari N.Ursolic acid regulates aging process through enhancing of metabolic sensor proteins level[J].Biomed Pharmacother,2016,82:8-14.
9 Zhang MF,Shen YQ.Research progress on lipidemic regulation and antiobesity effects of oleanolic acid and ursolic acid[J].Drug Eval Res (药物评价研究),2015,38(1):90-97.
10 Zhang J,Wu SY,Wang L,et al.Effects of usolic acid on fat metabolism in high-fat-fed induced insulin resistance rats[J].Chin Arch Tradit Chin Med(中华中医药学刊),2010,28:975-978.
11 Zeng L,Tang WJ,Yin JJ,et al.Experimental study on improving steatosis of hepatocytes with ursolic acid[J].Chin Med Mater(中药材),2015,38:1049-1052.
12 Liu K,Czaja MJ.Regulation of lipid stores and metabolism by lipophagy[J].Cell Death & Differentiation,2013,20(1):3-11.
13 Feng Y,He D,Yao Z,et al.The machinery of macroautophagy[J].Cell Res,2014,24(1):24-41.
14 Sahni S,Merlot AM,et al.Gene of the month:BECN1[J].J Clin Pathol,2014,67:656-660.
15 Duran A,Amanchy R,Linares JF,et al.P62 is a key regulator of nutrient sensing in the mTORC1 pathway[J].Molecular Cell,2011,44:134-146.
16 Zhu YS,Liu JL.Cell autophagy and mitochondrial quality control with healthy aging[J].Prog Biochem Biophys (生物化学与生物物理进展),2014,41:247-256.
17 Fige MT,Osiewacz HD,Reichert AS.Quality control of mitochondria during aging:is there a good and a badside of mitochondrial dynamics[J].Bioessays,2013,35:314-322.
18 Yang CC,Chen D,Lee SS,et al.The dynamin-related protein DRP-1 and the insulin signaling pathway cooperate to modulate Caenorhabditis elegans longevity[J].Aging Cell,2011,10:724-728.
19 Vives-Bauza C,Zhou C,Huang Y,et al.PINK1-dependent recruitment of Parkin to mitochondria in mitophagy[J].Proc Natl Acad Sci USA,2010,107:378-383.
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