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| 过氧化物酶体增殖物激活受体γ调控微RNA-223水平改善高脂诱导胰岛素抵抗细胞糖吸收障碍 |
| 张雪1,2△,田毅君3△,王玉芳1,杨霖1,刘佳佳1,蔡邦兰1,薛晓成4* |
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(1. 海军军医大学(第二军医大学)附属公利医院, 上海市炎症与慢病管理人工智能实验室, 上海 200135;
2. 宁夏医科大学基础医学院, 银川 750000;
3. 海军军医大学(第二军医大学)第三附属医院泌尿外科, 上海 201805;
4. 海军军医大学(第二军医大学)附属公利医院耳鼻咽喉科, 上海 200135
△共同第一作者
*通信作者) |
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| 摘要: |
| 目的 观察过氧化物酶体增殖物激活受体γ（PPARγ）调控miRNA-223水平在高脂诱导的胰岛素抵抗中的作用。方法 Wistar雄性大鼠通过高脂饲料喂养，建立2型糖尿病动物模型并予以吡格列酮治疗，以qPCR、葡萄糖氧化酶-过氧化物酶法和ELISA法检测吡格列酮治疗前后2型糖尿病大鼠血清miRNA-223、血糖及4种炎症因子（IL-1β、IL-6、IL-8、TNF-α）水平的变化；以软脂酸诱导HepG2细胞胰岛素抵抗模型，上调或抑制PPARγ蛋白或miRNA-223水平后，以细胞葡萄糖吸收实验、蛋白质印迹法和qPCR法检测细胞葡萄糖吸收水平、细胞葡萄糖转运蛋白（葡萄糖转运蛋白1、2、4；胰岛素受体底物1、2）表达及miRNA-223水平的变化。结果 与治疗前相比，吡格列酮治疗7 d后大鼠空腹血糖水平下降、血清miRNA-223水平上升、血清炎症因子（IL-1β、IL-6、IL-8、TNF-α）相对表达量下降（P均<0.001）。经软脂酸处理24 h后，软脂酸组HepG2细胞PPARγ蛋白表达和miRNA-223水平均较空白对照组降低，且细胞葡萄糖吸收水平下降（P均<0.05）；上调PPARγ蛋白可改善软脂酸导致的细胞葡萄糖吸收水平下降、升高miRNA-223及炎症因子水平（P均<0.05）。抑制PPARγ表达可导致细胞葡萄糖吸收水平降低（P<0.05）、miRNA-223表达水平下降（P<0.05），但对正常状态下炎症因子水平无调节作用；过表达miRNA-223可改善软脂酸诱导的细胞葡萄糖吸收水平下降和炎症因子水平增高（P均<0.05），以及上调葡萄糖转运蛋白1、葡萄糖转运蛋白4、胰岛素受体底物1的表达（P均<0.05）；抑制miRNA-223对软脂酸诱导的细胞葡萄糖吸收水平及葡萄糖转运蛋白1、葡萄糖转运蛋白4、胰岛素受体底物1表达的作用与过表达miRNA-223效果相反（P均<0.05）。但过表达或抑制miRNA-223均对PPARγ表达无影响。结论 PPARγ通过调节miRNA-223水平改善高脂诱导的胰岛素抵抗细胞葡萄糖吸收。 |
| 关键词: 过氧化物酶体增殖物激活受体 微RNA-223 胰岛素抵抗 2型糖尿病 葡萄糖吸收 |
| DOI：10.16781/j.CN31-2187/R.20210797 |
| 投稿时间：2021-08-13修订日期：2021-12-07 |
| 基金项目:上海市浦东新区卫生科技发展委基金（PKJ2018-Y09），上海市浦东新区卫生系统优秀青年医学人才培养计划（PWRq2020-61，PWRq-2020-63），上海市卫生健康委员会青年项目（20204Y0144），上海市浦东新区重点亚专科项目（PWZy2020-06），上海市浦东新区临床特色学科项目（PWYts2021-15）. |
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| Role of peroxisome proliferators-activated receptor γ in improving glucose absorption disorders in high-fat induced insulin resistance cells by regulating microRNA-223 |
| ZHANG Xue1,2△,TIAN Yi-jun3△,WANG Yu-fang1,YANG Lin1,LIU Jia-jia1,CAI Bang-lan1,XUE Xiao-cheng4* |
(1. Gongli Hospital, Naval Medical University (Second Military Medical University), Shanghai Health Commission Key Lab of Artificial Intelligence (AI)-Based Management of Inflammation and Chronic Diseases, Shanghai 200135, China;
2. College of Basic Medicine Sciences, Ningxia Medical University, Yinchuan 750000, Ningxia Hui Autonomous Region, China;
3. Department of Urology, The Third Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 201805, China;
4. Department of Otolaryngology, Gongli Hospital, Naval Medical University (Second Military Medical University), Shanghai 200135, China
△Co-first authors.
* Corresponding author) |
| Abstract: |
| Objective To investigate the role of peroxisome proliferators-activated receptors γ (PPARγ) in hight-fat induced insulin resistance by regulating microRNA (miRNA)-223. Methods The rat model of type 2 diabetes mellitus was established by feeding high-fat diet and treated with pioglitazone. Serum miRNA-223, blood glucose and 4 inflammatory factors (interleukin[IL]-1β, IL-6, IL-8, and tumor necrosis factor α[TNF-α]) in type 2 diabetes mellitus rats before and after treatment were detected by quantitative polymerase chain reaction (qPCR), glucose oxidase peroxidase and enzyme-linked immunosorbent assay. In a model of insulin resistance in HepG2 cells induced by palmitic acid, the levels of PPARγ or miRNA-223 were upregulated or inhibited, and the level of cellular glucose absorbtion, expression of cell glucose transporters (glucose transporters 1, 2, 4; insulin receptor substrates 1, 2) and miRNA-223 level were detected by Cell glucose absorption experiment, Western blotting and qPCR, respectively. Results Compared with that before pioglitazone treatment, the fasting blood glucose level of rats and the expression of serum inflammatory factors (IL-1β, IL-6, IL-8, and TNF-α) were significantly decreased and the level of serum miRNA-223 was significantly increased after 7 d of pioglitazone treatment (all P<0.001). After 24-h treatment with palmitic acid, the expression of PPARγ protein, miRNA-223 level and glucose absorption level of HepG2 cells were significantly decreased (all P<0.05). Upregulation of PPARγ protein could ameliorate the decrease of cellular glucose absorption induced by palmitic acid and increase the levels of miRNA-223 and inflammatory factors (all P<0.05). Inhibition of PPARγ expression resulted in decreased glucose absorption (P<0.05) and decreased miRNA-223 level (P<0.05), but there was no regulatory effect on the level of inflammatory factors in normal cells. Overexpression of miRNA-223 could improve the palmitic acid-induced decrease of glucose absorption and the increase of inflammatory factors (both P<0.05), as well as up-regulate the expression of glucose transporters (glucose transporters 1, 4) and insulin receptor substrate 1 (all P<0.05). Inhibition of miRNA-223 had an opposite effect of overexpression of miRNA-223 on the level of PA-induced cellular glucose uptake and expression of glucose transporter protein 1, glucose transporter protein 4, and insulin receptor substrate 1 (all P<0.05). However, overexpression or inhibition of miRNA-223 did not affect the expression of PPARγ. Conclusion PPARγ can improve glucose absorption of insulin resistance cells induced by high fat by regulating miRNA-223. |
| Key words: peroxisome proliferators-activated receptor γ microRNA-223 insulin resistance type 2 diabetes glucose absorption |
