Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells

Min WANG; Ya-qi LI; Quan-tao MA; Chen-yue LIU; Chi ZHANG; Min-yi QIU; Cai-juan ZHANG; Ting WANG; Bao-sheng ZHAO

doi:10.13422/j.cnki.syfjx.20190128

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Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells

Pharmacology | 更新时间：2021-02-09

- Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 1, Pages: 135-140(2019)
- 作者机构：
  
  北京中医药大学　中药学院，　北京中医药研究院，北京　 100029
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190128
  CLC： R22;R24;R285;R285.5;TQ467.32
- Received：07 August 2018，
  
  Published Online：22 October 2018，
  
  Published：05 January 2019
- 稿件说明：
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Min WANG, Ya-qi LI, Quan-tao MA, et al. Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 135-140.
DOI：

Min WANG, Ya-qi LI, Quan-tao MA, et al. Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 135-140. DOI： 10.13422/j.cnki.syfjx.20190128.

摘要

目的：

观察桑叶含药血清对脂肪细胞株3T3-L1胰岛素抵抗（IR）模型葡萄糖消耗量及细胞活力的影响，筛选出桑叶含药血清的最佳浓度，并检测其对炎症因子含量的影响，探讨可能的作用机制。

方法：

取对数生长期3T3-L1前脂肪细胞，10 mg·L

－1

胰岛素（Ins），0.25 mmol·L

－1

地塞米松（DEX），0.5 mmol·L

－1

3-异丁基-1-甲基黄嘌呤（IBMX）诱导48 h后，10 mg·L

－1

Ins再次诱导48 h，待其分化为成熟脂肪细胞后，1 μmol·L

－1

DEX诱导96 h以建立IR细胞模型。桑叶水提物含药血清培养12，24，36，72 h后，采用葡萄糖氧化酶法检测桑叶含药血清培养后细胞葡萄糖的消耗量，采用噻唑蓝（MTT）比色法检测桑叶含药血清培养后IR模型的细胞活力，酶联免疫吸附测定法（ELISA）检测桑叶含药血清对细胞肿瘤坏死因子-

（TNF-

）含量的影响，蛋白免疫印迹法（Western blot）测定桑叶含药血清对胰岛素信号通路胰岛素受体（InsR），胰岛素受体底物（IRS），磷酸化IRS1（p-IRS1），葡萄糖转运蛋白4（GLUT4）蛋白表达的影响。

结果：

桑叶含药血清可显著提高IR细胞葡萄糖消耗量（

0.01），增强细胞活力（

0.01），降低炎症因子TNF-

的含量（

0.01），调节胰岛素信号通路下游蛋白的表达量（

0.05，

0.01）。

结论：

桑叶可明显改善3T3-L1细胞IR状态，其作用机制可能与抑制TNF-

表达、促进胰岛素信号通路蛋白的表达有关。

Abstract

Objective:

To observe effect of Mori Folium-containing serum on glucose consumption and cell activity of fat cell line 3T3-L1 insulin resistance (IR) model

in order to screen out the optimal concentration of drug-containing serum

detect effect of Mori Folium on the content of inflammatory factors

and explore the possible mechanism.

Method:

3T3-L1 preadipocytes in logarithmic growth phase were selected

and induced with 10 mg·L

-1

insulin (Ins)

0.25 mmol·L

-1

dexamethasone (DEX) and 0.5 mmol·L

-1

3-isobutyl-methylxanthine(IBMX) for 48 h and then with 10 mg·L

-1

Ins for 48 h. After the cells were differentiated into mature adipocytes

they were induced with 1 μmol·L

-1

DEX for 96 h to establish IR model. Glucose content in the supernatant of cells was detected by glucose oxidase after serum containing Mori Folium cultured for 12

72 h. Methyl-thiazdyl-tetrazolium(MTT) was used to detect the effect of serum containing Mori Folium on IR cells activity. The content of tumor necrosis factor-

(TNF-

) was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile

the effects of inflammatory factors on the expressions of insulin signaling pathway proteins insulin receptor (InsR)

insulin receptor substrate (IRS)

p-IRS1 and glucose transporter 4 (GLUT4) were determined by Western blot.

Result:

Serum containing Mori Folium could significantly increase the glucose consumption rate and cell activity of IR cells (

0.01)

reduce the content of TNF-

(

0.01)

and regulate the expression of insulin signaling pathway proteins (

0.05

0.01).

Conclusion:

Mori Folium can significantly improve IR status of 3T3-L1 cells

and its mechanism may be related to inhibiting TNF-

and promoting the expressions of insulin signaling pathway proteins.

关键词

Keywords

references

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