您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报 (医学版) ›› 2023, Vol. 61 ›› Issue (11): 38-47.doi: 10.6040/j.issn.1671-7554.0.2023.0753

• • 上一篇    

NR4A1对毒胡萝卜素诱导C2C12肌管细胞糖代谢功能障碍的影响

芦兴晨1,强晔2,刘昕宇3,左丹4,姜子晗1,张玉超4,刘元涛5,马小莉2   

  1. 1.青岛大学附属青岛临床医学院 青岛市市立医院, 山东 青岛 266011;2.康复大学青岛医院(青岛市市立医院)内分泌中心, 山东 青岛 266011;3.烟台毓璜顶医院莱山分院临床营养科, 山东 烟台 264000;4.康复大学青岛医院(青岛市市立医院)医学杂志编辑部, 山东 青岛 266011;5.山东大学齐鲁医院内分泌科(青岛), 山东 青岛 266011
  • 发布日期:2023-12-12
  • 通讯作者: 马小莉. E-mail:maxl72@163.com; 刘元涛. E-mail:sduliuyuantao@126.com
  • 基金资助:
    山东省自然科学基金面上项目(ZR2022MH086);山东省医药卫生科技发展计划项目(202103060651);2021年度青岛市医药卫生科研计划项目(2021-WJZD005)

Effects of NR4A1 on thapsigargin-induced glucose metabolism dysfunction in C2C12 myotube cells

LU Xingchen1, QIANG Ye2, LIU Xinyu3, ZUO Dan4, JIANG Zihan1, ZHANG Yuchao4, LIU Yuantao5, MA Xiaoli2   

  1. 1. Qingdao Clinical Medical School of Qingdao University, Qingdao Municipal Hospital, Qingdao 266011, Shandong, China;
    2. Endocrine Center, Qingdao Hospital of Rehabilitation University(Qingdao Municipal Hospital), Qingdao 266011, Shandong, China;
    3. Department of Clinical Nutrition, Yantai Yuhuangding Hospital, Laishan Branch, Yantai 264000, Shandong, China;
    4. Editorial Office of Medical Journal, Qingdao Hospital of Rehabilitation University(Qingdao Municipal Hospital), Qingdao 266011, Shandong, China;
    5. Department of Endocrinology, Qilu Hospital of Shandong University(Qingdao), Qingdao 266011, Shandong, China
  • Published:2023-12-12

PDF (PC)

261

摘要： 目的 探讨NR4A1对毒胡萝卜素(TG)诱导小鼠C2C12肌管细胞糖代谢功能障碍的影响及机制。 方法 体外诱导C2C12细胞分化为成熟肌管细胞,将加入20 nmol/L胰岛素刺激10 min的细胞标记为胰岛素组,未做特殊处理的细胞标记为对照组,采用Western blotting法检测磷酯酰肌醇-3-激酶/蛋白激酶B(PI3K/AKT)信号通路、NR4A1蛋白水平。采用CCK8法检测TG浓度梯度处理的C2C12肌管细胞活力,建立糖代谢损伤细胞模型。将C2C12肌管细胞分为对照组、无水乙醇(EtOH)组、TG组,分别处理后加入胰岛素刺激,采用葡萄糖试剂盒检测葡萄糖消耗水平,采用Western blotting法检测NR4A1、p-PI3K、p-AKT蛋白表达。采用慢病毒感染并筛选,获得对照与稳定过表达NR4A1的C2C12肌管细胞,并进行TG处理,分为Ad-NC组、Ad-NC+EtOH组、Ad-NC+TG组和Ad-NR4A1+TG组。分化成熟后加入胰岛素刺激,检测葡萄糖消耗水平,检测NR4A1、p-PI3K、p-AKT的表达。 结果 与对照组相比,胰岛素组C2C12肌管细胞PI3K/AKT通路磷酸化水平及NR4A1表达水平显著升高(P<0.01)。CCK-8结果显示,TG浓度大于1 μmol/L时,细胞活力显著下降(P<0.01)。与EtOH组相比,TG组葡萄糖消耗减少,NR4A1表达水平下降,PI3K/AKT蛋白磷酸化水平受到抑制(P均< 0.01)。胰岛素刺激C2C12肌管细胞,与Ad-NC+TG组相比,Ad-NR4A1+TG组葡萄糖消耗增加(P<0.05)、PI3K/AKT信号通路显著活化(P<0.05)。 结论 TG导致C2C12肌管细胞糖代谢功能障碍。NR4A1对TG诱导的C2C12肌管细胞糖代谢障碍具有改善作用,其机制可能通过PI3K/AKT信号途径发挥作用。

关键词: NR4A1, C2C12肌管细胞, 磷酯酰肌醇-3-激酶/蛋白激酶B信号通路, 毒胡萝卜素, 糖代谢

Abstract: Objective To explore the effects and mechanism of NR4A1 on thapsigargin(TG)-induced glucose metabolism dysfunction in C2C12 myotube cells. Methods C2C12 cells were induced to differentiate into mature myotube cells in vitro. Cells stimulated with 20 nmol/L insulin for 10 minutes were labeled as insulin group, while cells without special treatment were labeled as control group. The phosphatidyl inositol-3-kinase/protein kinase B(PI3K/AKT)signaling pathway and protein expression of NR4A1 were determined with Western blotting. The viability of C2C12 myotube cells treated with TG concentration gradient was detected with CCK8. After a glucose metabolism impaired cell model was established, the cells were divided into control group, ethanol(EtOH)group, and TG group. After treatment, insulin was added, then glucose consumption level was measured with a glucose assay kit. The protein expressions of NR4A1, p-PI3K and p-AKT were detected with Western blotting. After the cells were infected with lentivirus and screened, those with stable overexpression of NR4A1 were obtained and subjected to TG treatment. After that, the cells were divided into Ad-NC group, Ad-NC+TG group, and Ad-NR4A1+TG group. After differentiation and maturation, insulin was added to detect the glucose consumption level and the expressions of NR4A1, p-PI3K and p-AKT. Results Compared with the control group, the insulin group showed significantly higher phosphorylation of the PI3K/AKT pathway and NR4A1 expression(P<0.01). CCK-8 results showed that the cell viability significantly decreased after 1 μmol/L of TG treatment(P<0.01). Compared with the EtOH group, the TG group showed decreased glucose consumption, reduced NR4A1 expression, and inhibited PI3K/AKT phosphorylation(P<0.01). Compared with the Ad-NC+TG group, the Ad-NR4A1+TG group showed an increase in glucose consumption with insulin stimulation(P<0.05), and significant activation of PI3K/KT signaling pathway(P<0.05). Conclusion TG induces glucose metabolism dysfunction in C2C12 myotube cells, while NR4A1 can improve such dysfunction, which may be mediated through the PI3K/AKT signaling pathway.

Key words: NR4A1, C2C12 myotube cells, Phosphatidyl inositol-3-kinase/protein kinase B signaling pathway, Thapsigargin, Glucose metabolism

中图分类号: 

  • R574
[1] Sun H, Saeedi P, Karuranga S, et al. IDF Diabetes Atlas: global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045 [J]. Diabetes Res Clin Pract, 2022, 183: 109119. doi: 10.1016/j.diabres.2021.109119.
[2] Zheng Y, Ley SH, Hu FB. Global aetiology and epidemiology of type 2 diabetes mellitus and its complications [J]. Nat Rev Endocrinol, 2018, 14(2): 88-98.
[3] Kim C, Lee J, Kim MB, et al. Hypoglycemic effect of whole grain diet in C57BL/KsJ-db/db mice by activating PI3K/Akt and AMPK pathways [J]. Food Sci Biotechnol, 2019, 28(3): 895-905.
[4] Guo X, Sun W, Luo G, et al. Panax notoginseng saponins alleviate skeletal muscle insulin resistance by regulating the IRS1-PI3K-AKT signaling pathway and GLUT4 expression [J]. FEBS Open Bio, 2019, 9(5): 1008-1019.
[5] 衣雪洁, 孙玉霞, 姚婷婷, 等. 急、慢性运动对2型糖尿病大鼠脂肪PI3K/AKT/GLUT4通路的影响[J]. 中国应用生理学杂志, 2020, 36(1): 12-16. YI Xuejie, SUN Yuxia, YAO Tingting, et al. Effects of acute and chronic exercise on fat PI3K/AKT/GLUT4 signal pathway in type 2 diabetic rats [J]. Chinese Journal of Applied Physiology, 2020, 36(1): 12-16.
[6] 张素素, 印遇龙, 孙嘉, 等. NR4A1调控系统糖脂代谢的分子机制[J]. 生命科学, 2021, 33(4): 449-459. ZHANG Susu, YIN Yulong, SUN Jia, et al. NR4A1 regulates systemic glucose and lipid metabolism [J]. Chinese Bulletin of Life Sciences, 2021, 33(4): 449-459.
[7] Chao LC, Zhang ZD, Pei LM, et al. Nur77 coordinately regulates expression of genes linked to glucose metabolism in skeletal muscle [J]. Mol Endocrinol, 2007, 21(9): 2152-2163.
[8] Kanzleiter T, Preston E, Wilks D, et al. Overexpression of the orphan receptor Nur77 alters glucose metabolism in rat muscle cells and rat muscle in vivo [J]. Diabetologia, 2010, 53(6): 1174-1183.
[9] Chen F, Yu Y, Tian H, et al. Nur77 is involved in the regulation of obesity-related lower muscle mass by promoting Pten degradation [J]. FASEB J, 2023, 37(8): e23083.
[10] Wang Y, Gao Y, Wang Y, et al. GDNF promotes the proliferation and osteogenic differentiation of jaw bone marrow mesenchymal stem cells via the Nr4a1/PI3K/Akt pathway [J]. Cell Signal, 2023, 108: 110721. doi: 10.1016/j.cellsig.2023.110721.
[11] Shi Z, To SKY, Zhang S, et al. Hypoxia-induced Nur77 activates PI3K/Akt signaling via suppression of Dicer/let-7i-5p to induce epithelial-to-mesenchymal transition [J]. Theranostics, 2021, 11(7): 3376-3391.
[12] Hedrick E, Lee SO, Safe S. The nuclear orphan receptor NR4A1 regulates β1-integrin expression in pancreatic and colon cancer cells and can be targeted by NR4A1 antagonists [J]. Mol Carcinog, 2017, 56(9): 2066-2075.
[13] Evans PL, McMillin SL, Weyrauch LA, et al. Regulation of skeletal muscle glucose transport and glucose metabolism by exercise training [J]. Nutrients, 2019, 11(10): 2432.
[14] Wu HZ, Ballantyne CM. Skeletal muscle inflammation and insulin resistance in obesity [J]. J Clin Investig, 2017, 127(1): 43-54.
[15] Rudnicki MA, Jaenisch R. The MyoD family of transcription factors and skeletal myogenesis [J]. BioEssays, 1995, 17(3): 203-209.
[16] Lith SC, de Vries CJM. Nuclear receptor Nur77: its role in chronic inflammatory diseases [J]. Essays Biochem, 2021, 65(6): 927-939.
[17] Lee SO, Li X, Khan S, et al. Targeting NR4A1(TR3)in cancer cells and tumors [J]. Expert Opin Ther Targets, 2011, 15(2): 195-206.
[18] Duszka K, Bogner-Strauss JG, Hackl H, et al. Nr4a1 is required for fasting-induced down-regulation of Pparγ2 in white adipose tissue [J]. Mol Endocrinol, 2013, 27(1): 135-149.
[19] Pan X, Liu B, Chen S, et al. Nr4a1 as a myogenic factor is upregulated in satellite cells/myoblast under proliferation and differentiation state [J]. Biochem Biophys Res Commun, 2019, 513(3): 573-581.
[20] Kanzleiter T, Wilks D, Preston E, et al. Regulation of the nuclear hormone receptor nur77 in muscle: influence of exercise-activated pathways in vitro and obesity in vivo [J]. Biochim Biophys Acta, 2009, 1792(8): 777-782.
[21] Chao LC, Wroblewski K, Zhang Z, et al. Insulin resistance and altered systemic glucose metabolism in mice lacking Nur77 [J]. Diabetes, 2009, 58(12): 2788-2796.
[22] Petersen MC, Shulman GI. Mechanisms of insulin action and insulin resistance [J]. Physiol Rev, 2018, 98(4): 2133-2223.
[23] Boucher J, Kleinridders A, Kahn CR. Insulin receptor signaling in normal and insulin-resistant states [J]. CSH Perspect Biol, 2014, 6(1): a009191.
[24] Qureshi FM, Dejene EA, Corbin KL, et al. Stress-induced dissociations between intracellular calcium signaling and insulin secretion in pancreatic islets [J]. Cell Calcium, 2015, 57(5-6): 366-375.
[25] Niu G, Ye T, Qin L, et al. Orphan nuclear receptor TR3/Nur77 improves wound healing by upregulating the expression of integrin β4 [J]. FASEB J, 2015, 29(1): 131-140.
[26] Lammi J, Aarnisalo P. FGF-8 stimulates the expression of NR4A orphan nuclear receptors in osteoblasts [J]. Mol Cell Endocrinol, 2008, 295(1-2): 87-93.
[27] Mohankumar K, Lee J, Wu CS, et al. Bis-indole-derived NR4A1 ligands and metformin exhibit NR4A1-dependent glucose metabolism and uptake in C2C12 cells [J]. Endocrinology, 2018, 159(5): 1950-1963.
[28] Kovalovsky D, Refojo D, Liberman AC, et al. Activation and induction of NUR77/NURR1 in corticotrophs by CRH/cAMP: involvement of calcium, protein kinase A, and MAPK pathways [J]. Mol Endocrinol, 2002, 16(7): 1638-1651.
[1] 刘敏,张玉超,马小莉,刘昕宇,孙露,左丹,刘元涛. 孤核受体NR4A1在H2O2诱导小鼠肾脏足细胞损伤中的作用[J]. 山东大学学报 (医学版), 2022, 60(5): 16-21.
[2] 扈艳雯,王志媛,郁万江,赵蕙琛,韩合理,徐志鹏,马红,张玉超,刘元涛. 52例肥胖患者脂肪分布与代谢综合征及糖代谢指标的相关性[J]. 山东大学学报 (医学版), 2020, 1(8): 101-106.
[3] 孙凤娇,李艳,王雨心,张天然,吴虹,高希宝. 膳食结构与硒对大鼠糖代谢的作用[J]. 山东大学学报 (医学版), 2020, 58(2): 36-43.
[4] 王美建,侯新国,梁凯,陈丽. 山东省城市中老年人群血脂现状调查[J]. 山东大学学报（医学版）, 2017, 55(5): 70-75.
[5] 程向登,梁爽,胡艳艳,乔玉,王凤雪,李桂梅. Prader-Willi综合征患儿下丘脑-垂体-靶腺轴功能及代谢的改变[J]. 山东大学学报（医学版）, 2017, 55(3): 107-111.
[6] 于斐,刘少壮,仲明惟,黄鑫,焦杰,胡三元,于文滨. 基于GC-TOF-MS的结直肠癌代谢组学差异分析[J]. 山东大学学报（医学版）, 2016, 54(7): 60-68.
[7] 赵琰,郑亚冰,闫新峰,张虎,常晓天. 筛选类风湿关节炎中糖代谢关键基因的探讨[J]. 山东大学学报（医学版）, 2016, 54(3): 30-35.
[8] 庄泳, 李栋, 付金秋, 时庆, 鞠秀丽. 儿童B系急性淋巴细胞白血病树突状细胞的生物学特性[J]. 山东大学学报（医学版）, 2014, 52(11): 60-64.
[9] 梁凯1，侯新国1，王新华2，宋允胜2，井庆平3，曲勇4，孙宇1，宋君1，董晓琳1，许聿新1，李振作1，付春莉1，李文娟1，王云霞1，矫磊1，曹立君1，尹小芳1，范琳琳1，陈丽1. 人体肥胖测量参数对糖代谢异常风险的预测价值研究[J]. 山东大学学报(医学版), 2012, 50(9): 59-62.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发