脂肪组织蛋白质快速提取新方案
栏目:技术    日期:2021-02-04    来源:admin

以往认为脂肪组织是单纯能量储存器官,但随着脂肪组织在能量控制、炎症反应和免疫应答方面重要作用的验证,其已被确认也是一个内分泌器官。脂肪组织不仅响应来自传统内分泌系统和中枢神经系统的传入信号,还表达和分泌具有重要功能的细胞因子,包括瘦素(Leptin)、炎性细胞因子、脂联素(adiponectin)、补体成分、纤溶酶原激活物抑制剂-1(PAI-1)、肾素-血管紧张素系统蛋白和抵抗素(Resistin)等。

 

脂肪组织中除了脂肪细胞外,还有干细胞、前脂肪细胞、巨噬细胞、嗜中性粒细胞、淋巴细胞和内皮细胞等。这些不同类型的细胞与能量稳态的维持密切相关。对脂肪组织中各类细胞增殖和分化调节的研究以及对不同细胞类型之间相互关系的研究极为重要。

 

2014年加州大学Ajay Chawla教授在Cell杂志上发表了热中性条件下小鼠模型中激活二型固有淋巴细胞(ILC2)可以调节白色脂肪组织中米色脂肪生成(棕色化),米色脂肪生成不仅需要IL-4(嗜酸性粒细胞分泌)和儿茶酚胺(巨噬细胞分泌),同时还需要ILC2和PDGFR阳性前脂肪细胞的参与。该研究让我们进一步了解了炎症与肥胖之间的关系,对于促进白色脂肪棕色化治疗肥胖和糖尿病具有重要意义。

Min-Woo Lee, Activated Type 2 Innate Lymphoid Cells Regulate Beige Fat Biogenesis, Cell,2014

 

同年,莫纳什大学Tony Tiganis教授在Cell杂志上发表了关于瘦素和胰岛素作用于POMC神经元促进白色脂肪棕色化的相关研究成果——瘦素和胰岛素能够协同作用于下丘脑的POMC神经元,使POMC神经元参与到白色脂肪棕色化过程,通过增加能量消耗以限制体重增加。该研究证明了POMC神经元可以整合瘦素和胰岛素信号,促进白色脂肪棕色化维持能量平衡。

(瘦素是一种由脂肪组织分泌的激素,作用位于中枢神经系统的受体从而调控生物行为以及新陈代谢,通过负反馈机制来调控生物体的能量平衡以及体重)。

 

2019年马克斯普朗克生物化学研究所Matthias Mann教授在Cell Metabolism杂志发表了针对棕色脂肪细胞的分泌蛋白质组学,并通过比较分析人类棕色和白色脂肪细胞分泌蛋白图谱,揭示了EPDR1是一种新型的棕色脂肪因子,为了进一步研究EPDR1的生物学功能,研究者分别在细胞系和小鼠体内进行了EPDR1敲除,比较了突变体与野生型之间的代谢差异或功能差别。研究发现,EPDR1敲除的小鼠表现出较低的耗氧量,并调控整个生物个体的能量代谢功能。

 

Atul S. Deshmukh, Proteomics-Based Comparative Mapping of the Secretomes of Human Brown and White Adipocytes Reveals EPDR1 as a Novel Batokine,Cell Metabolism,2019

 

近年来围绕脂肪样品的相关研究越来越多,但脂肪组织中脂肪细胞具有高脂质和低蛋白的特性:

脂肪细胞主要分为两类——白色脂肪细胞(存储能量)、棕色脂肪细胞(脂质氧化产生热量)。白色脂肪细胞属于单泡脂肪细胞,90%的细胞体积被脂滴占据,细胞质呈“圆环”样,而细胞核呈“半月形”,细胞器较少;棕色脂肪细胞属于多泡脂肪细胞,线粒体大而富集。

且脂肪组织中还包含着结缔组织,神经组织,基质血管成分等,故脂肪组织中提取到足量且符合要求的蛋白质具有极大的挑战性。

 


                                        图片来源:zhihu

Invent公司研发的柱式法动物脂肪细胞和组织总蛋白提取试剂盒(Cat#AT-022),离心管柱及配套裂解液可以快速有效的从脂肪组织或脂肪细胞中将蛋白和油脂分离,提高蛋白质得率及质量,可获取不同类型的蛋白质(天然蛋白/变性蛋白)满足下游不同实验的需求。



MinuteTM Total Protein Extraction Kit for Adipose Tissue/Cultured cells (AT-022) 已发表文章:

1. Wang, N., Liu, Y., Ma, Y., & Wen, D. (2017). High-intensity interval versus moderate-intensity continuous training: Superior metabolic benefits in diet-induced obesity mice. Life Sciences.

2. Qiantao Zheng(2017),Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. www.pnas.org/cgi/doi/10.1073/pnas.1707853114

3. Li etal (2017).High Efficiency Protein Extraction from Adipose Tissues3. BioTechniques 63:191 (November 2017) DOI: 10.2144/000114601

4. Micah L. Battson, Dustin M Lee, Dillon K Jarrell, Shuofei Hou, Kayl E Ecton, Tiffany L Weir, and Christopher L Gentile(2017).Suppression of Gut Dysbiosis Reverses Western Diet-Induced Vascular Dysfunction. AM J PHYSIOL-ENDOC M doi.org/10.1152/ajpendo.00187.2017

5. Guizong Chen, Qin Li ,Yanping Luo, Tao Liu, Shaolong ,Zhou Er Pan, Lixia Peng(2018).Effect of Notoginsenoside R1 on autologous adipose graft in rats.spandidos-publications.com. doi.org/10.3892/mmr.2018.8596

6. Tianxing Chen,Xiaoying Cheng,Yun Wang,Wu Yin(2018).Toosendanin inhibits adipogenesis by activating Wnt/β-catenin signaling.SCiENtifiC REPorTS | (2018) 8:4626 | DOI:10.1038/s41598-018-22873-x

7. N Wang ,Y Liu ,Y Ma ,D Wen (2018).Hydroxytyrosol ameliorates insulin resistance by modulating endoplasmic reticulum stress and prevents hepatic steatosis in diet-induced obesity mice. Journal of Nutritional Biochemistry , doi.org/10.1016/j.jnutbio.2018.03.018 

8. Weijie Du, Arnold Piek, E. Marloes Schouten, Cees W.A. van de Kolk, Christian Mueller, Alexandre Mebazaa, Adriaan A. Voors, Rudolf A. de Boer,Herman H.W. Sillje(2018).Plasma levels of heart failure biomarkers are primarily a reflection of extracardiac production.Theranostics . doi: 10.7150/thno.26055

9. Melissa Hann(2018).Anti-inflammatory Activity of Isomaltodextrin in a C57BL/6NCrl Mouse Model with Lipopolysaccharide-induced Low-grade Chronic Inflammation. http://hdl.handle.net/10214/14124.

10. Raffael Ott etal (2018). Reduced insulin receptor expression and altered DNA methylation in fat tissues and blood of women with GDM and offspring.The Journal of Clinical Endocrinology & Metabolism, jc.2018-01659, doi.org/10.1210/jc.2018-01659.

11. Lei Liu, Ningning Wang etal (2018).Saponins from Boussingaultia gracilis prevent obesity and related metabolic impairments in diet-induced obese mice. Food Funct. DOI: 10.1039/C8FO01264D.

12. SJ Yue (2018).Berberine alleviates insulin resistance by reducing peripheral branched-chain amino acids.Am J Physiol Endocrinol Metab. 2018 Nov 13. doi: 10.1152/ajpendo.00256.2018.

13. Shah, A. P., Johnson, M. D., Fu, X., Boersma, G. J., Shah, M., Wolfgang, M. J., ... & Baraban, J. M. (2019). Deletion of translin (Tsn) induces robust adiposity and hepatic steatosis without impairing glucose tolerance. International Journal of Obesity.doi.org/10.1038/s41366-018-0315-7.

14. Li M et al(2019). TNF-α Upregulates IKKε Expression via the Lin28B/let-7a Pathway to Induce Catecholamine Resistance in Adipocytes.Obesity (Silver Spring). Doi: 10.1002/oby.22434. 

15. Jahandideh, F., de Campos Zani, S. C., Son, M., Proctor, S. D., Davidge, S. T., Chan, C. B., & Wu, J. (2019). Egg white hydrolysate enhances insulin sensitivity in high fat diet induced insulin resistant rats via AKT activation. British Journal of Nutrition, 1-25. DOI10.1017/S0007114519000837.

16. Zhang, F., Li, D., Wu, Q., Sun, J., Guan, W., Hou, Y., ... & Wang, J. (2019). Prepartum body conditions affect insulin signaling pathways in postpartum adipose tissues in transition dairy cows. Journal of Animal Science and Biotechnology, 10(1), 38.doi.org/10.1186/s40104-019-0347-4 

17. Wang, N., Ma, Y., Liu, Z., Liu, L., Yang, K., Wei, Y., ... & Wen, D. (2019). Hydroxytyrosol prevents PM2. 5-induced adiposity and insulin resistance by restraining oxidative stress related NF-κB pathway and modulation of gut microbiota in a murine model. Free Radical Biology and Medicine, 141, 393-407. Zani, S. (2019). 

18. Campos Zani, S. C(2019).Exploratory study of egg white hydrolysate mechanisms of action in insulin sensitive tissues of high fat diet-induced insulin resistant rats. doi.org/10.7939/r3-vpcv-w240

19. Vianello, E., Dozio, E., Bandera, F., Froldi, M., Micaglio, E., Lamont, J., ... & Schmitz, G. (2020). Correlative Study on Impaired Prostaglandin E2 Regulation in Epicardial Adipose Tissue and its Role in Maladaptive Cardiac Remodeling via EPAC2 and ST2 Signaling in Overweight Cardiovascular Disease Subjects. International Journal of Molecular Sciences, 21(2), 520.




close
close