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[大鼠骨髓间充质干细胞条件培养基对棕榈酸诱导的HepG2细胞胰岛素抵抗的影响]

[Effects of conditioned media for rat bone marrow-derived mesenchymal stem cells on palmitic acid-induced insulin resistance in HepG2 cells].

作者信息

Sun Xiaoya, Hao Haojie, Han Weidong, Mu Yiming

机构信息

Department of Endocrinology, Chinese PLA General Hospital, Beijing 100853, China; Email:

出版信息

Zhonghua Nei Ke Za Zhi. 2015 May;54(5):439-44.

Abstract

OBJECTIVE

To study the effect of conditioned media for rat bone marrow mesenchymal stem cells (BMSCs-CM) on palmitic acid (PA)-induced insulin resistance (IR) in HepG2 cells and its underlying molecular mechanisms.

METHODS

HepG2 cells were treated with or without BMSCs-CM and L-DMEM in the presence or absence of PA.Glucose utilization in HepG2 cells were detected with PAS, glucose and glycogen measurements. Western blotting was used to assess the expression of phospho-insulin receptor substrate (p-IRS), phosphatidylinositol 3-kinase (PI3K) and p-AKT.

RESULTS

(1) Incubation of HepG2 cells with 0.25 mmol/L PA for 24 hours significantly increased the glucose concentration and decreased the glycogen content (P<0.05) in the media. (2) Treatment with BMSCs-CM significantly ameliorated the glucose and glycogen alteration in cells pretreated with PA (P<0.05), however, no obvious effect of BMSCs-CM on the cell glucose and glycogen production. (3) BMSCs-CM treatment also increased protein expression of p-IRS, PI3K and p-AKT in PA incubated HapG2 cells (P<0.05). The effect of BMSCs-CM on PI3K and p-AKT expression could be mimicked upon addition of 740Y-P, a PI3K agonist, but abolished by LY294002, a PI3K specific inhibitor.

CONCLUSIONS

BMSCs-CM could improve the insulin sensitivity in HepG2 cells pretreated with PA through upregulation of insulin signaling component expression.

摘要

目的

研究大鼠骨髓间充质干细胞条件培养基(BMSCs-CM)对棕榈酸(PA)诱导的HepG2细胞胰岛素抵抗(IR)的影响及其潜在分子机制。

方法

在有或无PA的情况下,用或不用BMSCs-CM和L-DMEM处理HepG2细胞。用PAS法检测HepG2细胞中的葡萄糖利用情况,并测量葡萄糖和糖原含量。采用蛋白质免疫印迹法评估磷酸化胰岛素受体底物(p-IRS)、磷脂酰肌醇3激酶(PI3K)和p-AKT的表达。

结果

(1)用0.25 mmol/L PA孵育HepG2细胞24小时后,培养基中的葡萄糖浓度显著升高,糖原含量降低(P<0.05)。(2)用BMSCs-CM处理可显著改善PA预处理细胞中的葡萄糖和糖原变化(P<0.05),然而,BMSCs-CM对细胞葡萄糖和糖原生成无明显影响。(3)BMSCs-CM处理还增加了PA孵育的HapG2细胞中p-IRS、PI3K和p-AKT的蛋白表达(P<0.05)。添加PI3K激动剂740Y-P可模拟BMSCs-CM对PI3K和p-AKT表达的影响,但PI3K特异性抑制剂LY294002可消除这种影响。

结论

BMSCs-CM可通过上调胰岛素信号成分表达来改善PA预处理的HepG2细胞的胰岛素敏感性。

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