[1]童国相 王莎 高国应 何一伟 李琼.Apelin-13对3T3-L1前体脂肪细胞增殖 与分化的影响[J].国际内分泌代谢杂志,2019,39(05):302-306.[doi:10.3760/cma.j.issn.1673-4157.2019.05.004]
　Tong Guoxiang,Wang Sha,Gao Guoying,et al.Effects of Apelin-13 on proliferation and differentiation of 3T3-L1 preadipocytes[J].International Journal of Endocrinology and Metabolism,2019,39(05):302-306.[doi:10.3760/cma.j.issn.1673-4157.2019.05.004]
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Apelin-13对3T3-L1前体脂肪细胞增殖 与分化的影响()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
39
期数:
2019年05期
页码:
302-306
栏目:
论著
出版日期:
2019-09-20

文章信息/Info

Title:
Effects of Apelin-13 on proliferation and differentiation of 3T3-L1 preadipocytes
作者:
童国相 王莎 高国应 何一伟 李琼
长沙医学院附属第一医院内分泌科 410219
Author(s):
Tong Guoxiang;  Wang Sha;  Gao Guoying;  He Yiwei;  Li Qiong
Department of Endocrinology, The First Affiliated Hospital of Changsha Medical University, Changsha 410219, China
关键词:
Apelin;  3T3-L1前体脂肪细胞;  细胞增殖;  细胞分化;  过氧化物酶体增殖物活化受体γ
Keywords:
Apelin;  3T3-L1 preadipocytes;  Cell proliferation;  Cell differentiation;  Peroxisome proliferator activated receptor γ
DOI:
10.3760/cma.j.issn.1673-4157.2019.05.004
摘要:
目的 探讨Apelin-13对3T3-L1前体脂肪细胞增殖与分化的影响及其可能的作用机制。方法 体外培养3T3-L1前体脂肪细胞。取对数生长期细胞,分别予以不同浓度Apelin-13进行干预,四甲基偶氮唑盐(MTT)法检测Apelin-13对3T3-L1前体脂肪细胞增殖的影响。“经典鸡尾酒法”诱导3T3-L1前体脂肪细胞分化。将对数生长期细胞分为实验组和对照组。实验组分别于诱导分化第2、4、6、8天更换培养液的同时,予以细胞存活率抑制作用最强浓度的Apelin-13进行干预。对照组不做处理。于诱导分化第8天,根据油红O染色、脂质和甘油三酯含量检测细胞分化程度。于诱导分化第2、4、6、8天,分别采用RT-PCR和Western印迹检测过氧化物酶体增殖物活化受体γ(PPARγ)mRNA和蛋白表达的变化。结果 Apelin-13浓度越高,干预时间越长,3T3-L1前体脂肪细胞存活率越低。以100 μmol/L Apelin-13干预96 h后,3T3-L1前体脂肪细胞存活率最低。经Apelin-13干预的3T3-L1前体脂肪细胞脂滴、脂质和甘油三酯含量均明显少于对照组(t=4.526、5.353、4.827,P均<0.05)。与对照组相比,诱导分化第6、8天,经Apelin-13干预的3T3-L1前体脂肪细胞PPARγ mRNA和蛋白表达量显著下降(t=4.962、5.416、4.734、5.627, P均<0.05)。结论 Apelin-13呈浓度和时间依赖性抑制3T3-L1前体脂肪细胞的增殖,并抑制3T3-L1前体脂肪细胞分化过程中脂滴的形成,减少脂质聚集和甘油三酯含量。其作用机制可能与通过抑制PPARγ的表达有关。
Abstract:
Objective To explore the effects of Apelin-13 on the proliferation and differentiation of 3T3-L1 preadipocytes and its possible mechanisms.Methods 3T3-L1 preadipocytes were cultured in vitro. The logarithmic growth cells were treated with different concentrations of Apelin-13. The effects of Apelin-13 on the proliferation of 3T3-L1 preadipocytes were detected by methyl thiazolyl tetrazolium(MTT)assay. The "classical cocktail" method was used to induce the differentiation of 3T3-L1 preadipocytes. The logarithmic growth cells were divided into experimental group and control group. In experimental group, the culture mediums were replaced on the 2nd, 4th, 6th and 8th day of differentiation induction, respectively; at the same time cells were intervened with Apelin-13 in a concentration with the strongest inhibitory effect on cell livability. Cells in control group received no intervention. On the 8th day of differentiation induction, the degree of differentiation was detected by Oil red O staining, lipid and triglyceride assessment. Peroxisome proliferator activated receptor γ(PPARγ)mRNA and protein expression were detected by RT-PCR and Western blotting on the 2nd, 4th, 6th and 8th day of differentiation induction, respectively.Results The cell livability of 3T3-L1 preadipocytes was decreased along with the increase of the concentration as well as the intervention time of Apelin-13. The lowest livability of 3T3-L1 preadipocytes was observed after 96 hours of intervention with 100 μmol/L Apelin-13. Compared with control group, the lipid droplets, lipid and triglyceride content of 3T3-L1 preadipocytes treated with Apelin-13 were significantly lower(t=4.526, 5.353, 4.827, all P<0.05). The expression of PPARγ mRNA and protein in 3T3-L1 preadipocytes treated with Apelin-13 were decreased significantly on the 6th and 8th day of differentiation(t=4.962, 5.416, 4.734, 5.627, all P<0.05), compared with control group.Conclusions Apelin-13 inhibits the proliferation of 3T3-L1 preadipocytes in a concentration and time dependent manner. In addition, Apelin-13 could inhibit the formation of lipid droplets, and reduce lipid accumulation and triglyceride content during the differentiation of 3T3-L1 preadipocytes. The possible mechanism is related to inhibit the expression of PPARγ.

参考文献/References:

[1] Shin K,Chapman NA,Sarker M,et al.Bioactivity of the putative apelin proprotein expands the repertoire of apelin receptor ligands[J].Biochim Biophys Acta Gen Subj,2017,1861(8):1901-1912.DOI:10.1016/j.bbagen.2017.05.017.
[2] 肖罡,唐圣松,涂剑. Apelin/APJ系统对能量代谢及其相关疾病调控的影响[J]. 中国生物化学与分子生物学报,2018, 34(5): 481-485. DOI:10.13865/j.cnki.cjbmb.2018.05.04.
[3] Sekerci R,Acar N,Tepekoy F,et al.Apelin/APJ expression in the heart and kidneys of hypertensive rats[J].Acta Histochem,2018,120(3):196-204.DOI:10.1016/j.acthis.2018.01.007.
[4] 程玉文,张振洲,余俊贤,等.ACE2/Apelin信号在心血管系统中的调控作用及相关靶向治疗药物[J].药学进展,2017,41(7):483-489.
[5] Coquerel D,Chagnon F,Sainsily X,et al.ELABELA improves cardio-renal outcome in fatal experimental septic shock[J].Crit Care Med,2017,45(11):e1139-e1148.DOI:10.1097/CCM.0000000000002639.
[6] Akhondali Z,Badavi M,Dianat M,et al.Co-administration of apelin and T4 protects inotropic and chronotropic changes occurring in hypothyroid rats[J].Arq Bras Cardiol,2015,105(3):235-240.DOI:10.5935/abc.20150086.
[7] Adam F,Khatib AM,Lopez JJ,et al.Apelin: an antithrombotic factor that inhibits platelet function[J].Blood,2016,127(7):908-920.DOI:10.1182/blood-2014-05-578781.
[8] Guo M,Chen F,Lin T,et al.Apelin-13 decreases lipid storage in hypertrophic adipocytes in vitro through the upregulation of AQP7 expression by the PI3K signaling pathway[J].Med Sci Monit,2014,20:1345-1352.DOI:10.12659/MSM.890124.
[9] Sawane M,Kajiya K,Kidoya H,et al.Apelin inhibits diet-induced obesity by enhancing lymphatic and blood vesselintegrity[J].Diabetes,2013,62(6):1970-1980.DOI:10.2337/db12-0604.
[10] 童国相,刘俊,陈珂,等.脂肪细胞特异性核酸适体对肥胖小鼠脂肪代谢的影响[J].中国现代医学杂志,2014,24(26):12-17.
[11] Sabry RN,El Wakeel MA,El-Kassas GM,et al.Serum apelin: a new marker of early atherosclerosis in children with type 1 diabetes mellitus[J].Open Access Maced J Med Sci,2018,6(4):613-617.DOI:10.3889/oamjms.2018.144.
[12] Boal F,Timotin A,Roumegoux J,et al.Apelin-13 administration protects against ischaemia/reperfusion-mediated apoptosis through the FoxO1 pathway in high-fat diet-induced obesity[J].Br J Pharmacol,2016,173(11):1850-1863.DOI:10.1111/bph.13485.
[13] Yang L,Su T,Lv D,et al.ERK1/2 mediates lung adenocarcinoma cell proliferation and autophagy induced by apelin-13[J].Acta Biochim Biophys Sin(Shanghai),2014,46(2):100-111.DOI:10.1093/abbs/gmt140.
[14] Bülbül M,Sinen O,Gök M,et al.Apelin-13 inhibits gastric motility through vagal cholinergic pathway in rats[J].Am J Physiol Gastrointest Liver Physiol,2018,314(2):G201-G210.DOI:10.1152/ajpgi.00223.2017.
[15] Montanari T,Posˇc'ic' N,Colitti M.Factors involved in white-to-brown adipose tissue conversion and in thermogenesis: a review[J].Obes Rev,2017,18(5):495-513.DOI:10.1111/obr.12520.
[16] Petrus P,Mejhert N,Corrales P,et al.Transforming growth factor-β3 regulates adipocyte number in subcutaneous white adipose tissue[J].Cell Rep,2018,25(3):551-560.e5.DOI:10.1016/j.celrep.2018.09.069.
[17] Than A,He HL,Chua SH,et al.Apelin enhances brown adipogenesis and browning of white adipocytes[J].J Biol Chem,2015,290(23):14679-14691.DOI:10.1074/jbc.M115.643817.
[18] Ni Y,Ji C,Wang B,et al.A novel pro-adipogenesis factor abundant in adipose tissues and over-expressed in obesity acts upstream of PPARγ and C/EBPα[J].J Bioenerg Biomembr,2013,45(3):219-228.DOI:10.1007/s10863-012-9492-6.
[19] Tung EWY,Ahmed S,Peshdary V,et al.Firemaster 550 and its components isopropylated triphenyl phosphate and triphenyl phosphate enhance adipogenesis and transcriptional activity of peroxisome proliferator activated receptor(Pparγ)on the adipocyte protein 2(aP2)promoter[J].PLoS One,2017,12(4):e0175855. DOI:10.1371/journal.pone.0175855.
[20] Briot A,Decaunes P,Volat F,et al.Senescence alters PPARγ(peroxisome proliferator-activated receptor gamma)-dependent fatty acid handling in human adipose tissue microvascular endothelial cells and favors inflammation[J].Arterioscler Thromb Vasc Biol,2018,38(5):1134-1146. DOI:10.1161/ATVBAHA.118.310797.

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备注/Memo

备注/Memo:
通信作者:童国相,Email:togtog2008@126.com
Corresponding author: Tong Guoxiang,Email: togtog2008@126.com
基金项目:湖南省教育厅科学研究项目(16C0169)
Fund program:The Scientific Research Project of Hunan Provincial Education Department(16C0169)
更新日期/Last Update: 2019-09-20