黄连温胆汤含药血清对胰岛素抵抗脂肪细胞改善作用的蛋白质组学研究
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摘要
目的:探索黄连温胆汤含药血清改善脂肪细胞胰岛素抵抗的可能机制。方法:将3T3-L1成纤维细胞分为空白组、模型组和中药组,对模型组和中药组3T3-L1成纤维细胞进行诱导分化,构建胰岛素抵抗3T3-L1脂肪细胞模型。空白组、模型组分别给予空白血清刺激,中药组给予黄连温胆汤含药血清。使用TMT标记、液相色谱-质谱联用分析,并运用GO分析和KEGG pathway分析方法对获得的差异表达蛋白进行生物信息学分析。结果:本实验中3组样本共鉴定到4 982个蛋白质,其中4238个蛋白质具有定量信息。模型组与空白组间上调蛋白210个,下调蛋白399个;中药组和空白组间上调蛋白221个,下调蛋白98个;中药组和模型组组间比较上调蛋白310个,下调蛋白142个。GO分析结果示中药组/模型组间差异表达上调蛋白显著富集于DNA复制、核染色质、蛋白结合等分类中,下调蛋白显著富集于氧化还原、脂肪酸β氧化等分类。KEGG pathway富集分析示,差异表达上调蛋白富集于10条通路中,富集较显著的通路为DNA复制、细胞循环、不匹配修复等通路;差异表达下调蛋白富集于25条通路中,富集较显著的通路为过氧化物酶体、脂肪酸降解、脂肪酸代谢等通路。结论:黄连温胆汤含药血清改善胰岛素抵抗效果确切,其机制是通过多靶点、多通路协同、整体调控作用完成的。
Objective: To explore possible mechanisms of HuanglianWendan decoction drug-containing serum to improve insulin resistance. Methods:3T3-L1 fibroblasts were divided into control group, model group and TCM group. 3T3-L1 fibroblasts in model group and TCM group were induced to differentiate, and the 3T3-L1 adipocyte model of insulin resistance was constructed. The control group and model group were given normal serum stimulation, while the TCM group was treatment with drug-containing serum. Finally, proceed bioinformatics analysis of the acquired differential expression protein by using TMT marking, liquid chromatography-mass(LC-MS) spectrometry, GO analysis and KEGG pathway analysis. Results: 4982 proteins are identified in three group of samples in this experiment, 4238 of them contain quantitative information. 210 up-regulate and 399 down-regulate proteins exist in model group and control group; 221 up-regulate and 98 down-regulate ones are identified in TCM group and control group; 310 up-regulate and 142 down-regulate proteins come to light with TCM group and control group. The result shows that the differentially expressed proteins were mainly distributed in GO terms, such as DNA replication, nucleoplasm, protein binding terms of TCM group and control group. KEGG pathway enrichment analysis shows that, differentially expressed up-regulate proteins were enriched in 10 pathways, significant enriched pathways are DNA replication, cell cycle and mismatch repair, etc. Besides, differentially expressed down-regulate proteins were enriched in 25 pathways, significant enriched pathways are peroxisome, fatty acid degradation and metabolism process, etc. Conclusion: It is clear that HuanglianWendan decoction drug-containing serum could improve insulin resistance by multi-targeting, multi-pathway synergy, and overall regulation.
Objective: To explore possible mechanisms of HuanglianWendan decoction drug-containing serum to improve insulin resistance. Methods:3T3-L1 fibroblasts were divided into control group, model group and TCM group. 3T3-L1 fibroblasts in model group and TCM group were induced to differentiate, and the 3T3-L1 adipocyte model of insulin resistance was constructed. The control group and model group were given normal serum stimulation, while the TCM group was treatment with drug-containing serum. Finally, proceed bioinformatics analysis of the acquired differential expression protein by using TMT marking, liquid chromatography-mass(LC-MS) spectrometry, GO analysis and KEGG pathway analysis. Results: 4982 proteins are identified in three group of samples in this experiment, 4238 of them contain quantitative information. 210 up-regulate and 399 down-regulate proteins exist in model group and control group; 221 up-regulate and 98 down-regulate ones are identified in TCM group and control group; 310 up-regulate and 142 down-regulate proteins come to light with TCM group and control group. The result shows that the differentially expressed proteins were mainly distributed in GO terms, such as DNA replication, nucleoplasm, protein binding terms of TCM group and control group. KEGG pathway enrichment analysis shows that, differentially expressed up-regulate proteins were enriched in 10 pathways, significant enriched pathways are DNA replication, cell cycle and mismatch repair, etc. Besides, differentially expressed down-regulate proteins were enriched in 25 pathways, significant enriched pathways are peroxisome, fatty acid degradation and metabolism process, etc. Conclusion: It is clear that HuanglianWendan decoction drug-containing serum could improve insulin resistance by multi-targeting, multi-pathway synergy, and overall regulation.
引文
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14 Avecilla V,Doke M,Felty Q.Contribution of inhibitor of DNA Binding/Differentiation-3and endocrine disrupting chemicals to pathophysiological aspects of chronic disease[J].Biomed Res Int,2017,2017:6307109.
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18 Doktorova M,Zwarts I,Zutphen TV,et al.Intestinal PPARδprotects against diet-induced obesity,insulin resistance and dyslipidemia[J].Sci Rep,2017,7(1):846.
19 Vazquez-Carrera M.Unraveling the effects of PPARβ/δon insulin resistance and cardiovascular disease[J].Trends Endocrinol Metab,2016,27(5):319-334.
20 Kang S,Tsai LT,Rosen ED.Nuclear mechanisms of insulin resistance[J].Trends Cell Biol,2016,26(5):341-351.
21 Liu JJ,Lu W,Shi BM,et al.Peroxisomal regulation of redox homeostasis and adipocyte metabolism[J].Redox Biol,2019,24:101167.
2刘紫君,韩宇博,彭鹏,等.黄连温胆汤含药血清改善小鼠脂肪细胞胰岛素抵抗的机制探讨[J].山东医药,2018,58(30):14-18.
3罗新新,朱水兰,黎宇,等.3T3-L1脂肪细胞胰岛素抵抗模型的建立、优化及分子指标鉴定[J].中药新药与临床药理,2018,29(2):225-231.
4马伯艳,李云凤,张风丽,等.近20年黄连温胆汤防治糖尿病及其并发症研究进展[J].辽宁中医杂志,2019,46(3):660-663.
5隋艳波,刘莉.黄连温胆汤治疗代谢综合征的临床疗效观察[J].中西医结合心脑血管病杂志,2015,13(5):581-582.
6刘洪双,刘莉,隋艳波.加减黄连温胆汤治疗代谢综合征(痰湿蕴结证)的临床研究[J].中国中医急症,2016,25(10):1837-1839,1850.
7仲维莉,邹国良,刘莉,等.黄连温胆汤加减对代谢综合征大鼠骨骼肌细胞葡萄糖转运子-4的影响[J].中医药学报,2014,42(4):66-68.
8 Peng YH,Sun LX,Jia Z,et al.Predicting proteinDNA binding free energy change upon missense mutations using modified MM/PBSA approach:SAMPDIwebserver[J].Bioinformatics,2018,34(5):779-786.
9 Maciejczyk M,.Zebrowska E,Zalewska A,et al.Redox balance,antioxidant Defense,and oxidative damage in the hypothalamus and cerebral cortex of rats with high fat diet-induced insulin resistance[J].Oxid Med Cell Longev,2018,2018:6940515.
10 Acosta-Monta1o P,García-Gonzlez V.Effects of dietary fatty acids in pancreatic beta cell metabolism,implications in homeostasis[J].Nutrients,2018,10(4):E393.
11 Cen J,Sargsyan E,Bergsten P.Fatty acids stimulate insulin secretion from human pancreatic islets at fasting glucose concentrations via mitochondria-dependent andindependent mechanisms[J].NutrMetab(Lond),2016,13(1):59.
12 Jeˇzek P,Jab。urek M,HolendovB,etal.Fatty acidstimulated insulin secretion vs.lipotoxicity[J].Molecules,2018,23(6):E1483.
13 Kanehisa M,Sato Y,Furumichi M,et al.New approach for understanding genome variations in KEGG[J].Nucleic Acids Res,2019,47(D1):D590-D595.
14 Avecilla V,Doke M,Felty Q.Contribution of inhibitor of DNA Binding/Differentiation-3and endocrine disrupting chemicals to pathophysiological aspects of chronic disease[J].Biomed Res Int,2017,2017:6307109.
15 Chiefari E,Foti DP,Sgarra R,et al.Transcriptional regulation of glucose metabolism:the emerging role of the HMGA1chromatin factor[J].Front Endocrinol(Lausanne),2018,9:357.
16 Costa DK,Huckestein BR,Edmunds LR,et al.Reduced intestinal lipid absorption and body weight-independent improvements in insulin sensitivity in high-fat diet-fed Park2knockout mice[J].Am J Physiol Endocrinol Metab,2016,311(1):E105-E116.
17 Williams JA,Ni HM,Ding YF,et al.Parkin regulates mitophagy and mitochondrial function to protect against alcohol-induced liver injury and steatosis in mice[J].Am J PhysiolGastrointest Liver Physiol,2015,309(5):G324-G340.
18 Doktorova M,Zwarts I,Zutphen TV,et al.Intestinal PPARδprotects against diet-induced obesity,insulin resistance and dyslipidemia[J].Sci Rep,2017,7(1):846.
19 Vazquez-Carrera M.Unraveling the effects of PPARβ/δon insulin resistance and cardiovascular disease[J].Trends Endocrinol Metab,2016,27(5):319-334.
20 Kang S,Tsai LT,Rosen ED.Nuclear mechanisms of insulin resistance[J].Trends Cell Biol,2016,26(5):341-351.
21 Liu JJ,Lu W,Shi BM,et al.Peroxisomal regulation of redox homeostasis and adipocyte metabolism[J].Redox Biol,2019,24:101167.
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