红景天苷通过激活3T3-L1前脂肪细胞Nrf2/HO-1信号通路及下调脂肪生成转录因子表达来抑制脂肪生成
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摘要
目的:研究红景天苷对3T3-L1前脂肪细胞增殖分化及Nrf2/HO-1信号通路和脂肪生成转录因子的影响。方法:按文献介绍的方法诱导3T3-L1前脂肪细胞分化,然后分别用红景天苷(50和100μM)、吡格列酮(100μM)及红景天苷(100μM)与HO-1抑制剂ZnPP(3μM)共同处理3T3-L1细胞7天,分别于2、5、7天后进行胞内脂质含量测定,7天后进行胞内TG含量检测,实时定量PCR检测细胞中PPARγ、C/EBPα、SREBP-1c、aP2、adiponectin mRNA表达, Western blot检测胞浆和胞核中Nrf2及细胞中HO-1蛋白表达。结果:红景天苷(50和100μM)可显著抑制胞内脂质的积聚和TG生成,降低脂肪生成相关转录因子PPARγ、C/EBPα、SREBP-1c、aP2、adiponectin mRNA表达,抑制Nrf2核易位,上调HO-1蛋白表达,且量效关系明显;当与ZnPP联用后,上述作用被明显削弱。结论:红景天苷对3T3-L1前脂肪细胞分化和脂肪生成具有显著的抑制作用,其作用机制可能与其激活Nrf2/HO-1信号通路及抑制PPARγ、C/EBPα、SREBP-1c、aP2、adiponectin等脂肪生成转录因子的表达有关。
Objective: To investigate the effect of salidroside on proliferation and differentiation of 3T3-L1 preadipocytes,Nrf2/HO-1 signaling pathway and adipogenic transcription factors. Methods: 3 T3-L1 preadipocytes were induced to differentiate by the method described in the literature. Salidroside( 50 and 100μM),pioglitazone( 100μM),salidroside( 100μM) and HO-1 inhibitor ZnP P( 3μM) were used to cotreat 3T3-L1 cells for 7 days. Intracellular lipid content was measured after 2,5 and 7 days,and intracellular TG content was detected after7 days. Real-time PCR was applied to detect the mRNA expressions of PPARγ,C/EBPα,SREBP-1c,aP2 and adiponectin. Protein expressions of cytoplasm Nrf2,nucleus Nrf2 and HO-1 in 3T3-L1 preadipocytes were detected by western blotting. Results: Salidroside( 50 and100μM) significantly inhibited intracellular lipid accumulation and TG production,decreased the mRNA expressions of Fat-producing transcription factors PARgam,C/EBPa,SREBP-1c,aP2 and adiponectin,it also inhibited protein expressions of Nrf2 nuclear translocation,and up-regulate the expression of HO-1,in dose-dependent manner. When combined with ZnP P,the above effects were obviously weakened.Conclusion: Salidroside has the significant inhibitory effect on the differentiation and adipogenesis of 3 T3-L1 preadipocytes. Its mechanism may be related to activating Nrf2/HO-1 signaling pathway and inhibiting the expressions of adipogenic transcription factors such as PPARγ,C/EBPα,SREBP-1c,aP 2,adiponectin.
Objective: To investigate the effect of salidroside on proliferation and differentiation of 3T3-L1 preadipocytes,Nrf2/HO-1 signaling pathway and adipogenic transcription factors. Methods: 3 T3-L1 preadipocytes were induced to differentiate by the method described in the literature. Salidroside( 50 and 100μM),pioglitazone( 100μM),salidroside( 100μM) and HO-1 inhibitor ZnP P( 3μM) were used to cotreat 3T3-L1 cells for 7 days. Intracellular lipid content was measured after 2,5 and 7 days,and intracellular TG content was detected after7 days. Real-time PCR was applied to detect the mRNA expressions of PPARγ,C/EBPα,SREBP-1c,aP2 and adiponectin. Protein expressions of cytoplasm Nrf2,nucleus Nrf2 and HO-1 in 3T3-L1 preadipocytes were detected by western blotting. Results: Salidroside( 50 and100μM) significantly inhibited intracellular lipid accumulation and TG production,decreased the mRNA expressions of Fat-producing transcription factors PARgam,C/EBPa,SREBP-1c,aP2 and adiponectin,it also inhibited protein expressions of Nrf2 nuclear translocation,and up-regulate the expression of HO-1,in dose-dependent manner. When combined with ZnP P,the above effects were obviously weakened.Conclusion: Salidroside has the significant inhibitory effect on the differentiation and adipogenesis of 3 T3-L1 preadipocytes. Its mechanism may be related to activating Nrf2/HO-1 signaling pathway and inhibiting the expressions of adipogenic transcription factors such as PPARγ,C/EBPα,SREBP-1c,aP 2,adiponectin.
引文
[1]Hadrich F,Sayadi S.Apigetrin inhibits adipogenesis in 3T3-L1 cells by downregulating PPARγand CEBP-α.Lipids Health Dis,2018,17(1)∶95.doi:10.1186/s12944-018-0738-0.
[2]Yesmin Simu S,Ahn S,Castro-Aceituno V,et al.Ginsenoside Rg5:Rk1 exerts an anti-obesity effect on 3T3-L1 cell Line by the downregulation of PPARγand CEBPα.Iran J Biotechnol,2017,15(4)∶252~259.
[3]Lee S G,Lee Y J,Jang M H,et al.Panax ginseng leaf extracts exert anti-obesity effects in high-fat diet-induced obese rats.Nutrients,2017,9(9).pii:E999.doi:10.3390/nu9090999.
[4]Yun J W.Possible anti-obesity therapeutics from nature-A review.Phytochemistry,2010,71(14-15)∶1625~1641.
5国家药典委员会.中国药典(一部).北京:中国医药科技出版社,2015,154~155.
[6]Zhu Y,Zhang Y J,Liu W W,et al.Salidroside suppresses HUVECs cell injury induced by oxidative stress through activating the Nrf2 signaling pathway.Molecules,2016,21(8).pii∶E1033.
[7]李新明,高忠东,李群,等.红景天提取物对3T3-L1脂肪细胞增殖及细胞分化的影响.安徽农业科学,2015,43(5)∶25~26,139.
[8]王树海,王文健,汪雪峰,等.红景天苷对3T3-L1脂肪细胞糖代谢及细胞分化的影响.中西医结合学报,2004,2(3)∶193~195.
[9]Zheng T,Yang X Y,Wu D,et al.Salidroside ameliorates insulin resistance through activation of a mitochondria-associated AMPK/PI3K/Akt/GSK3βpathway.Br J Pharmacol,2015,172(13)∶3284~3301.
[10]周继刚,王海燕,李小妹,等.红景天苷通过调节Nrf2/HO-1和PPARγ/CEBPα信号通路抑制高脂诱导的大鼠肥胖.三峡大学学报(自然科学版),2019,41(3)∶109~113.
[11]Lee M S,Shin Y J,Jung S Y,et al.The inhibitory effects of tartary buckwheat ectracts on adipogenesis and inflammatory response.Molecules,2017,22(7).pii∶E1160.doi:10.3390/molecules22071160.
[12]白彩虹,邹坤,贺海波,等.吉祥草乙酸乙酯部位对人宫颈癌Caski细胞抑制作用及COX-2基因表达与BCl-2蛋白家族关系的研究.中药药理与临床,2013,29(6)∶45~50.
[13]Yang J W,Sung J Y,Kim Y W,et al.Inhibitory effects of butein on adipogenesis through upregulation of the Nrf2/HO-1 pathway in 3T3-L1 adipocyte.Prev Nutr Food Sci,2017,22(4)∶306~311.
[14]Singer K,Lumeng C N.The initiation of metabolic inflammation in childhood obesity.J Clin Invest,2017,127(1)∶65~73.
[15]Morrison M C,Kleemann R.Role of macrophage migration inhibitory factor in obesity,insulin resistance,type 2 diabetes,and associated hepatic co-morbidities:a comprehensive review of human and rodent studies.Front Immunol,2015,(6):308.
[16]Geeraerts X,Bolli E,Fendt S M,et al.Macrophage metabolism as therapeutic target for cancer,atherosclerosis,and obesity.Front Immunol,2017,(8)∶289.
[17]Liou C J,Wu S J,Chen LC,et al.Acacetin from traditionally used Saussurea involucrata Kar.et Kir.suppressed adipogenesis in 3T3-L1 adipocytes and attenuated lipid accumulation in obese mice.Front Pharmacol,2017,(8)∶589.
[18]Zingg J M,Hasan S T,Meydani M.Molecular mechanisms of hypolipidemic e?ects of curcumin.Biofactors,2013,39(1)∶101~121.
[19]Huang H H,Chen G Z,Liao D,et al.The effects of resveratrol intervention on risk markers of cardiovascular health in overweight and obese subjects:a pooled analysis of randomized controlled trials.Obes Rev,2016,17(12)∶1329~1340.
[20]Pi J B,Leung L R,Xue P,et al.Deficiency in the nuclear factor E2-related factor-2 transcription factor results in impaired adipogenesis and protects against diet-induced obesity.J Biol Chem,2010,285(12)∶9292~9300.
[21]Kwak M K,Wakabayashi N,Kensler T W.Chemoprevention through the Keap1-Nrf2 signaling pathway by phase 2 enzyme inducers.Mutat Res,2004,555(1-2)∶133~148.
[22]Abo El-Magd N F,El-Mesery M,El-Karef A,et al.Glycyrrhizin ameliorates high fat diet-induced obesity in rats by activating Nr F2 pathway.Life Sci,2018,193∶159~170.
[23]Han M H,Jeong J S,Jeong J W,et al.Ethanol extracts of Aster yomena(Kitam.)Honda inhibit adipogenesis through the activation of the AMPKsignaling pathway in 3T3-L1 preadipocytes.Drug Discov Ther,2017,11(5)∶281~287.
[24]Rosen E D,Walkey C J,Puigserver P,et al.Transcriptional regulation of adipogenesis.Genes Dev,2000,14(11)∶1293~1307.
[25]Mota de SáP,Richard A J,Hang H,et al.Transcriptional regulation of adipogenesis.Compr Physiol,2017,7(2)∶635~674.
[26]刘丽,林庶如,李军,等.毛蕊异黄酮葡萄糖苷与葛根素及其配伍对3T3-L1前脂肪细胞胰岛素抵抗细胞模型的影响.中药药理与临床,2018,34(1)∶10~14.
[27]Pandit R,Beerens S,Adan R A H.Role of leptin in energy expenditure:The hypothalamic perspective.Am J Physiol Regul Integr Comp Physiol,2017,312(6)∶R938~R947.
[28]Nakamura M T,Yudell B E,Loor J J.Regulation of energy metabolism by long-chain fatty acids.Prog Lipid Res,2014,53∶124~144.
[29]Coles C A.Adipokines in healthy skeletal muscle and metabolic disease.Adv Exp Med Biol,2016,900∶133~160.
[2]Yesmin Simu S,Ahn S,Castro-Aceituno V,et al.Ginsenoside Rg5:Rk1 exerts an anti-obesity effect on 3T3-L1 cell Line by the downregulation of PPARγand CEBPα.Iran J Biotechnol,2017,15(4)∶252~259.
[3]Lee S G,Lee Y J,Jang M H,et al.Panax ginseng leaf extracts exert anti-obesity effects in high-fat diet-induced obese rats.Nutrients,2017,9(9).pii:E999.doi:10.3390/nu9090999.
[4]Yun J W.Possible anti-obesity therapeutics from nature-A review.Phytochemistry,2010,71(14-15)∶1625~1641.
5国家药典委员会.中国药典(一部).北京:中国医药科技出版社,2015,154~155.
[6]Zhu Y,Zhang Y J,Liu W W,et al.Salidroside suppresses HUVECs cell injury induced by oxidative stress through activating the Nrf2 signaling pathway.Molecules,2016,21(8).pii∶E1033.
[7]李新明,高忠东,李群,等.红景天提取物对3T3-L1脂肪细胞增殖及细胞分化的影响.安徽农业科学,2015,43(5)∶25~26,139.
[8]王树海,王文健,汪雪峰,等.红景天苷对3T3-L1脂肪细胞糖代谢及细胞分化的影响.中西医结合学报,2004,2(3)∶193~195.
[9]Zheng T,Yang X Y,Wu D,et al.Salidroside ameliorates insulin resistance through activation of a mitochondria-associated AMPK/PI3K/Akt/GSK3βpathway.Br J Pharmacol,2015,172(13)∶3284~3301.
[10]周继刚,王海燕,李小妹,等.红景天苷通过调节Nrf2/HO-1和PPARγ/CEBPα信号通路抑制高脂诱导的大鼠肥胖.三峡大学学报(自然科学版),2019,41(3)∶109~113.
[11]Lee M S,Shin Y J,Jung S Y,et al.The inhibitory effects of tartary buckwheat ectracts on adipogenesis and inflammatory response.Molecules,2017,22(7).pii∶E1160.doi:10.3390/molecules22071160.
[12]白彩虹,邹坤,贺海波,等.吉祥草乙酸乙酯部位对人宫颈癌Caski细胞抑制作用及COX-2基因表达与BCl-2蛋白家族关系的研究.中药药理与临床,2013,29(6)∶45~50.
[13]Yang J W,Sung J Y,Kim Y W,et al.Inhibitory effects of butein on adipogenesis through upregulation of the Nrf2/HO-1 pathway in 3T3-L1 adipocyte.Prev Nutr Food Sci,2017,22(4)∶306~311.
[14]Singer K,Lumeng C N.The initiation of metabolic inflammation in childhood obesity.J Clin Invest,2017,127(1)∶65~73.
[15]Morrison M C,Kleemann R.Role of macrophage migration inhibitory factor in obesity,insulin resistance,type 2 diabetes,and associated hepatic co-morbidities:a comprehensive review of human and rodent studies.Front Immunol,2015,(6):308.
[16]Geeraerts X,Bolli E,Fendt S M,et al.Macrophage metabolism as therapeutic target for cancer,atherosclerosis,and obesity.Front Immunol,2017,(8)∶289.
[17]Liou C J,Wu S J,Chen LC,et al.Acacetin from traditionally used Saussurea involucrata Kar.et Kir.suppressed adipogenesis in 3T3-L1 adipocytes and attenuated lipid accumulation in obese mice.Front Pharmacol,2017,(8)∶589.
[18]Zingg J M,Hasan S T,Meydani M.Molecular mechanisms of hypolipidemic e?ects of curcumin.Biofactors,2013,39(1)∶101~121.
[19]Huang H H,Chen G Z,Liao D,et al.The effects of resveratrol intervention on risk markers of cardiovascular health in overweight and obese subjects:a pooled analysis of randomized controlled trials.Obes Rev,2016,17(12)∶1329~1340.
[20]Pi J B,Leung L R,Xue P,et al.Deficiency in the nuclear factor E2-related factor-2 transcription factor results in impaired adipogenesis and protects against diet-induced obesity.J Biol Chem,2010,285(12)∶9292~9300.
[21]Kwak M K,Wakabayashi N,Kensler T W.Chemoprevention through the Keap1-Nrf2 signaling pathway by phase 2 enzyme inducers.Mutat Res,2004,555(1-2)∶133~148.
[22]Abo El-Magd N F,El-Mesery M,El-Karef A,et al.Glycyrrhizin ameliorates high fat diet-induced obesity in rats by activating Nr F2 pathway.Life Sci,2018,193∶159~170.
[23]Han M H,Jeong J S,Jeong J W,et al.Ethanol extracts of Aster yomena(Kitam.)Honda inhibit adipogenesis through the activation of the AMPKsignaling pathway in 3T3-L1 preadipocytes.Drug Discov Ther,2017,11(5)∶281~287.
[24]Rosen E D,Walkey C J,Puigserver P,et al.Transcriptional regulation of adipogenesis.Genes Dev,2000,14(11)∶1293~1307.
[25]Mota de SáP,Richard A J,Hang H,et al.Transcriptional regulation of adipogenesis.Compr Physiol,2017,7(2)∶635~674.
[26]刘丽,林庶如,李军,等.毛蕊异黄酮葡萄糖苷与葛根素及其配伍对3T3-L1前脂肪细胞胰岛素抵抗细胞模型的影响.中药药理与临床,2018,34(1)∶10~14.
[27]Pandit R,Beerens S,Adan R A H.Role of leptin in energy expenditure:The hypothalamic perspective.Am J Physiol Regul Integr Comp Physiol,2017,312(6)∶R938~R947.
[28]Nakamura M T,Yudell B E,Loor J J.Regulation of energy metabolism by long-chain fatty acids.Prog Lipid Res,2014,53∶124~144.
[29]Coles C A.Adipokines in healthy skeletal muscle and metabolic disease.Adv Exp Med Biol,2016,900∶133~160.