[睾酮对3T3-L1脂肪细胞炎症因子表达的影响及其机制]

[The effects of testosterone on the expression of inflammatory factors in 3T3-L1 adipocytes and its mechanism].

作者信息

Su Chun-lin, Huang Hai-yan, Shen Zong-qi, Lin Jin-fang

机构信息

Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2009 Jun 2;89(21):1493-7.

PMID:19953904

Abstract

OBJECTIVE

To explore the effects of androgen upon the production of inflammatory Cultured factors in 3T3-L1 adipocytes and to investigate the mechanism at the molecular level.

METHODS

pre-adipocytes from 3T3-L1 cell line were induced to differentiate into adipocytes. Mature adipocytes were treated with testosterone at a concentration of 10(-9) to 10(-15) mol/L for either short (0 to 30 min) or long (12, 24 and 48 h) treatment course. Secretion of inflammatory factors, i.e., interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), in the culture medium were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of IL-6 and MCP-1 were determined by reverse transcriptase PCR. Phosphorylation of NF-KB was analyzed by Western blot with beta-actin as the reference gene. In another experiment, adipocytes were manipulated according to the same protocol except being pretreated with PDTC (inhibitor of NF-KB) for 2 h prior to the addition of testosterone. The results of two experiments were compared.

RESULTS

(1) The secretion of IL-6 and MCP-1 in the culture medium were higher in the testosterone-treated groups than in the control groups (P < 0.05). The highest concentration of IL-6 and MCP-1 were observed in the group treated with 10(-5) M testosterone for 24 h. The mRNA expression of IL-6 and MCP-1 were higher in the groups treated with testosterone for 12 h, especially with testosterone of 10(-5) mol/L; (2) With a short treatment course of testosterone, more NF-kappaB were phosphorylated than in control, especially with testosterone of 10(-5) mol/L. More NF-kappaB was phosphorylated following the 12 h testosterone treatment (10(-9), 10(-7) and 10(-5) mol/L), especially with a testosterone concentration of 10(-9) and 10(-5) mol/L. (3) When pretreated by NF-kappaB inhibitor and followed by 10(-5) mol/L testosterone for 24 h, the secretion of IL-6 and MCP-1 in the culture medium decreased significantly (P < 0.01). Likewise, when pretreated by NF-kappaB inhibitor and followed by 10(-5) M testosterone for 12 h, the mRNA expression of IL-6 and MCP-1 decreased significantly.

CONCLUSION

Within the certain scope, testosterone could increase the expression of inflammatory factors in adipocytes through the activation of NF-kappaB.

摘要

目的

探讨雄激素对3T3-L1脂肪细胞炎症因子产生的影响，并从分子水平研究其机制。

方法

将3T3-L1细胞系的前脂肪细胞诱导分化为脂肪细胞。成熟脂肪细胞用浓度为10(-9)至10(-15)mol/L的睾酮进行短期（0至30分钟）或长期（12、24和48小时）处理。通过酶联免疫吸附测定（ELISA）测定培养基中炎症因子白细胞介素-6（IL-6）和单核细胞趋化蛋白-1（MCP-1）的分泌。通过逆转录PCR测定IL-6和MCP-1的mRNA表达。以β-肌动蛋白为参照基因，通过蛋白质印迹法分析NF-κB的磷酸化。在另一实验中，脂肪细胞按相同方案处理，但在添加睾酮前先用PDTC（NF-κB抑制剂）预处理2小时。比较两个实验的结果。

结果

（1）睾酮处理组培养基中IL-6和MCP-1的分泌高于对照组（P<0.05）。在10(-5)M睾酮处理24小时的组中观察到IL-6和MCP-1的最高浓度。睾酮处理12小时的组中IL-6和MCP-1的mRNA表达较高，尤其是在10(-5)mol/L睾酮处理组；（2）睾酮短期处理时，磷酸化的NF-κB比对照组更多，尤其是在10(-5)mol/L睾酮处理组。睾酮处理12小时（10(-9)、10(-7)和10(-5)mol/L）后，磷酸化的NF-κB更多，尤其是在睾酮浓度为10(-9)和10(-5)mol/L时。（3）用NF-κB抑制剂预处理，然后用10(-5)mol/L睾酮处理24小时，培养基中IL-6和MCP-1的分泌显著降低（P<0.01）。同样，用NF-κB抑制剂预处理，然后用10(-5)M睾酮处理12小时，IL-6和MCP-1的mRNA表达显著降低。