电针对腹型肥胖大鼠肝脏脂代谢及Sirt1/PPARγ通路的影响
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摘要
目的:探讨电针对高脂饮食诱导的腹型肥胖大鼠肝脏脂代谢及沉默信息调节因子1(Sirts1)、过氧化物酶体增殖物激活受体γ(PPARγ)表达的影响。方法:SD雄性大鼠,随机分为空白组6只,造模组29只。造模组大鼠高脂饮食喂养12周,建立腹型肥胖模型。造模成功的12只腹型肥胖大鼠,随机分为模型组6只、针刺组6只,针刺组大鼠电针双侧"带脉"穴,2Hz/15Hz疏密波,电流强度1.5mA,每次20min,隔日1次,干预8周。每周测量体质量、腹围。针刺8周末,采用全自动生化仪检测血清总胆固醇(TC)、甘油三酯(TG)、谷丙转氨酶(ALT)、谷草转氨酶(AST),油红"O"染色法观察肝脏病理形态学变化,荧光定量PCR法检测肝组织Sirt1和PPARγ mRNA的表达,Western blot法检测肝组织Sirt1和PPARγ蛋白的表达水平。结果:与正常组比较,模型组的体质量、腹围明显增加(P<0.001,P<0.01),血清TC、TG、ALT、AST含量明显升高(P<0.01),肝脏脂质堆积明显增加,Sirt1 mRNA及蛋白的表达明显下降(P<0.05,P<0.01),PPARγ mRNA及蛋白的表达明显增加(P<0.01,P<0.05);与模型组比较,针刺组大鼠体质量、腹围明显减小(P<0.05,P<0.01),TC、TG、ALT、AST明显降低(P<0.05),肝脏脂质堆积明显减轻,肝组织Sirt1 mRNA及蛋白的表达显著增加(P<0.001,P<0.05),PPARγ mRNA及蛋白的表达明显降低(P<0.05,P<0.01)。结论:针刺可改善腹型肥胖大鼠肝脏脂质代谢,其机制可能与上调Sirt1、下调PPARγ的表达相关。
Objective To observe the impact of electroacupuncture(EA)on liver lipid metabolism and expression of hepatic sirtuin 1(Sirt1)and peroxisome proliferator activated receptorγ(PPARγ)of abdominal obese rats induced by high-fat diet.Methods Eighteen male SD rats were divided into blank control,model and EA groups(n=6 per group).The abdominal obesity model was established by feeding the rats with high-fat diet for 12 weeks.EA(2 Hz/15 Hz,1.5 mA)was applied to bilateral"Daimai"(GB26)for 20 min every time,once every other day for 8 weeks.Rats of the model group were also restrained for 20 min.The body mass and abdominal circumference were measured every week,and the contents of serum cholesterol(TC),triglyceride(TG),alanine transaminase(ALT),aspartate aminotransferase(AST)were detected by using an automated biochemical analyzer.Histopathological changes of the liver tissues were observed under microscope after oil red "O"staining.The expression of hepatic Sirt1 and PPARγ mRNAs and proteins were detected using quantitative real time PCR and Western blot,separately.Results After modeling,the body weight and abdominal circumference,and serum TC,TG,ALT and AST contents,and expression of hepatic PPARγ mRNA and protein were significantly increased(P<0.001,P<0.01,P<0.05),and the expression levels of hepatic Sirt1 mRNA and protein obviously down-regulated(P <0.05,P <0.01)in the model group.Following EA intervention,the increased body weight and abdominal circumference,and serum TC,TG,ALT and AST contents,and hepatic PPARγ mRNA and protein expression were remarkably suppressed(P<0.05,P<0.01),and the decreased hepatic Sirt1 mRNA and protein were remarkably up-regulated(P<0.001,P<0.05).The lipid droplets in hepatocytes were reduced in the EA group relevant to the model group.Conclusion EA intervention can significantly improve the liver lipid metabolism of abdominal obese rats,which is possibly related with its effect in up-regulating the expression of hepatic Sirt1 mRNA and protein,and in down-regulating the expression of hepatic PPARγ mRNA and protein.
Objective To observe the impact of electroacupuncture(EA)on liver lipid metabolism and expression of hepatic sirtuin 1(Sirt1)and peroxisome proliferator activated receptorγ(PPARγ)of abdominal obese rats induced by high-fat diet.Methods Eighteen male SD rats were divided into blank control,model and EA groups(n=6 per group).The abdominal obesity model was established by feeding the rats with high-fat diet for 12 weeks.EA(2 Hz/15 Hz,1.5 mA)was applied to bilateral"Daimai"(GB26)for 20 min every time,once every other day for 8 weeks.Rats of the model group were also restrained for 20 min.The body mass and abdominal circumference were measured every week,and the contents of serum cholesterol(TC),triglyceride(TG),alanine transaminase(ALT),aspartate aminotransferase(AST)were detected by using an automated biochemical analyzer.Histopathological changes of the liver tissues were observed under microscope after oil red "O"staining.The expression of hepatic Sirt1 and PPARγ mRNAs and proteins were detected using quantitative real time PCR and Western blot,separately.Results After modeling,the body weight and abdominal circumference,and serum TC,TG,ALT and AST contents,and expression of hepatic PPARγ mRNA and protein were significantly increased(P<0.001,P<0.01,P<0.05),and the expression levels of hepatic Sirt1 mRNA and protein obviously down-regulated(P <0.05,P <0.01)in the model group.Following EA intervention,the increased body weight and abdominal circumference,and serum TC,TG,ALT and AST contents,and hepatic PPARγ mRNA and protein expression were remarkably suppressed(P<0.05,P<0.01),and the decreased hepatic Sirt1 mRNA and protein were remarkably up-regulated(P<0.001,P<0.05).The lipid droplets in hepatocytes were reduced in the EA group relevant to the model group.Conclusion EA intervention can significantly improve the liver lipid metabolism of abdominal obese rats,which is possibly related with its effect in up-regulating the expression of hepatic Sirt1 mRNA and protein,and in down-regulating the expression of hepatic PPARγ mRNA and protein.
引文
[1]CARROLL J F,CHIAPA A L,RODRIQUEZ M,et al.Visceral fat,waist circumference,and BMI:impact of race/ethnicity[J].Obesity(Silver Spring),2008,16(3):600-607.
[2]KIM D,TOUROS A,KIM W R.Nonalcoholic fatty liver disease and metabolic syndrome[J].Clin Liver Dis,2018,22(1):133-140.
[3]GOODMAN Z D.The impact of obesity on liver histology[J].Clin Liver Dis,2014,18(1):33-40.
[4]MILIC′S,LULIC′D,TIMAC D.Non-alcoholic fatty liver disease and obesity:biochemical,metabolic and clinical presentations[J].World J Gastroenterol,2014,20(28):9330-9337.
[5]WANG Q Q,IMAM M U,YIDA Z,et al.Peroxisome proliferator-activated receptor gamma(PPARγ)as a target for concurrent management of diabetes and obesity-related cancer[J].Curr Pharm Des,2017,23(25):3677-3688.
[6]CAVE M C,CLAIR H B,HARDESTY J E,et al.Nuclear receptors and nonalcoholic fatty liver disease[J].Biochim Biophys Acta,2016,1859(9):1083-1099.
[7]MARIANI S,COSTANTINI D,LUBRANO C,et al.Circulating SIRT1inversely correlates with epicardial fat thickness in patients with obesity[J].Nutr Metab Cardiovasc Dis,2016,26(11):1033-1038.
[8]NGUYEN L T,CHEN H,ZAKY A,et al.SIRT1overexpression attenuates offspring metabolic and liver disorders as a result of maternal high-fat feeding[J].J Physiol(Lond),2019,597(2):467-480.
[9]PICARD F,KURTEV M,CHUNG N,et al.Sirt1promotes fat mobilization in white adipocytes by repressing PPAR-gamma[J].Nature,2004,429(6993):771-776.
[10]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[11]梁翠梅,胡慧,李媛媛.通调带脉法针刺治疗腹型肥胖疗效观察[J].针刺研究,2012,37(6):493-496.
[12]CHANDLER P C,VIANA J B,OSWALD K D,et al.Feeding response to melanocortin agonist predicts preference for and obesity from a high-fat diet[J].Physiol Behav,2005,85(2):221-230.
[13]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[14]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:329-332.
[15]HUI E,XU A M,BOYANG Y H,et al.Obesity as the common soil of non-alcoholic fatty liver disease and diabetes:role of adipokines[J].J Diabetes Investig,2013,4(5):413-425.
[16]KIM D,TOUROS A,KIM W R.Nonalcoholic fatty liver disease and metabolic syndrome[J].Clin Liver Dis,2018,22(1):133-140.
[17]RADMARD A R,POUSTCHI H,HASHEMI TAHERI AP,et al.Central obesity and liver iron content:a noninvasive assessment in general population by magnetic resonance imaging[J].Ann Nutr Metab,2016,69(3-4):181-189.
[18]梁翠梅,孙颂歌,胡慧.不同疗程针刺治疗腹型肥胖疗效初探[J].辽宁中医药大学学报,2018,20(8):169-172.
[19]沈凌宇,梁翠梅,杨文津,等.通调带脉法针刺治疗腹部肥胖型多囊卵巢综合征的随机对照研究[J].针刺研究,2018,43(4):255-259.
[20]CHOI S E,KWON S,SEOK S,et al.Obesity-linked phosphorylation of SIRT1by casein kinase 2inhibits its nuclear localization and promotes fatty liver[J].Mol Cell Biol,2017,37(15):e00006-e00017.
[21]PURUSHOTHAM A,SCHUG T T,XU Q,et al.Hepatocyte-specific deletion of SIRT1alters fatty acid metabolism and results in hepatic steatosis and inflammation[J].Cell Metab,2009,9(4):327-338.
[22]MORN-SALVADOR E,LPEZ-PARRA M,GARCA-ALONSO V,et al.Role for PPARγin obesity-induced hepatic steatosis as determined by hepatocyte-and macrophagespecific conditional knockouts[J].Faseb J,2011,25(8):2538-2550.
[23]LI Y,LAZAR M A.Differential gene regulation by PPAR-gamma agonist and constitutively active PPARgamma2[J].Mol Endocrinol,2002,16(5):1040-1048.
[24]PICARD F,KURTEV M,CHUNG N,et al.Sirt1promotes fat mobilization in white adipocytes by repressing PPAR-gamma[J].Nature,2004,429(6993):771-776.
[25]邹逸凡,马明珠,赵钊,等.隔药饼灸对高脂血症合并动脉粥样硬化兔肝脏过氧化酶体增殖物激活型受体γ、B类Ⅰ型清道夫受体蛋白及基因表达的影响[J].针刺研究,2018,43(2):86-91.
[26]SHIOMI Y,YAMAUCHI T,IWABU M,et al.A novel peroxisome proliferator-activated receptor(PPAR)αagonist and PPARγantagonist,Z-551,ameliorates high-fat diet-induced obesity and metabolic disorders in mice[J].J Biol Chem,2015,290(23):14567-14581.
[2]KIM D,TOUROS A,KIM W R.Nonalcoholic fatty liver disease and metabolic syndrome[J].Clin Liver Dis,2018,22(1):133-140.
[3]GOODMAN Z D.The impact of obesity on liver histology[J].Clin Liver Dis,2014,18(1):33-40.
[4]MILIC′S,LULIC′D,TIMAC D.Non-alcoholic fatty liver disease and obesity:biochemical,metabolic and clinical presentations[J].World J Gastroenterol,2014,20(28):9330-9337.
[5]WANG Q Q,IMAM M U,YIDA Z,et al.Peroxisome proliferator-activated receptor gamma(PPARγ)as a target for concurrent management of diabetes and obesity-related cancer[J].Curr Pharm Des,2017,23(25):3677-3688.
[6]CAVE M C,CLAIR H B,HARDESTY J E,et al.Nuclear receptors and nonalcoholic fatty liver disease[J].Biochim Biophys Acta,2016,1859(9):1083-1099.
[7]MARIANI S,COSTANTINI D,LUBRANO C,et al.Circulating SIRT1inversely correlates with epicardial fat thickness in patients with obesity[J].Nutr Metab Cardiovasc Dis,2016,26(11):1033-1038.
[8]NGUYEN L T,CHEN H,ZAKY A,et al.SIRT1overexpression attenuates offspring metabolic and liver disorders as a result of maternal high-fat feeding[J].J Physiol(Lond),2019,597(2):467-480.
[9]PICARD F,KURTEV M,CHUNG N,et al.Sirt1promotes fat mobilization in white adipocytes by repressing PPAR-gamma[J].Nature,2004,429(6993):771-776.
[10]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[11]梁翠梅,胡慧,李媛媛.通调带脉法针刺治疗腹型肥胖疗效观察[J].针刺研究,2012,37(6):493-496.
[12]CHANDLER P C,VIANA J B,OSWALD K D,et al.Feeding response to melanocortin agonist predicts preference for and obesity from a high-fat diet[J].Physiol Behav,2005,85(2):221-230.
[13]李媛媛,胡慧,梁翠梅,等.针刺“带脉”穴对代谢综合征大鼠体质量、血糖和脂代谢的影响[J].针刺研究,2014,39(3):202-206.
[14]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:329-332.
[15]HUI E,XU A M,BOYANG Y H,et al.Obesity as the common soil of non-alcoholic fatty liver disease and diabetes:role of adipokines[J].J Diabetes Investig,2013,4(5):413-425.
[16]KIM D,TOUROS A,KIM W R.Nonalcoholic fatty liver disease and metabolic syndrome[J].Clin Liver Dis,2018,22(1):133-140.
[17]RADMARD A R,POUSTCHI H,HASHEMI TAHERI AP,et al.Central obesity and liver iron content:a noninvasive assessment in general population by magnetic resonance imaging[J].Ann Nutr Metab,2016,69(3-4):181-189.
[18]梁翠梅,孙颂歌,胡慧.不同疗程针刺治疗腹型肥胖疗效初探[J].辽宁中医药大学学报,2018,20(8):169-172.
[19]沈凌宇,梁翠梅,杨文津,等.通调带脉法针刺治疗腹部肥胖型多囊卵巢综合征的随机对照研究[J].针刺研究,2018,43(4):255-259.
[20]CHOI S E,KWON S,SEOK S,et al.Obesity-linked phosphorylation of SIRT1by casein kinase 2inhibits its nuclear localization and promotes fatty liver[J].Mol Cell Biol,2017,37(15):e00006-e00017.
[21]PURUSHOTHAM A,SCHUG T T,XU Q,et al.Hepatocyte-specific deletion of SIRT1alters fatty acid metabolism and results in hepatic steatosis and inflammation[J].Cell Metab,2009,9(4):327-338.
[22]MORN-SALVADOR E,LPEZ-PARRA M,GARCA-ALONSO V,et al.Role for PPARγin obesity-induced hepatic steatosis as determined by hepatocyte-and macrophagespecific conditional knockouts[J].Faseb J,2011,25(8):2538-2550.
[23]LI Y,LAZAR M A.Differential gene regulation by PPAR-gamma agonist and constitutively active PPARgamma2[J].Mol Endocrinol,2002,16(5):1040-1048.
[24]PICARD F,KURTEV M,CHUNG N,et al.Sirt1promotes fat mobilization in white adipocytes by repressing PPAR-gamma[J].Nature,2004,429(6993):771-776.
[25]邹逸凡,马明珠,赵钊,等.隔药饼灸对高脂血症合并动脉粥样硬化兔肝脏过氧化酶体增殖物激活型受体γ、B类Ⅰ型清道夫受体蛋白及基因表达的影响[J].针刺研究,2018,43(2):86-91.
[26]SHIOMI Y,YAMAUCHI T,IWABU M,et al.A novel peroxisome proliferator-activated receptor(PPAR)αagonist and PPARγantagonist,Z-551,ameliorates high-fat diet-induced obesity and metabolic disorders in mice[J].J Biol Chem,2015,290(23):14567-14581.