脂联素对大鼠原代Kupffer细胞白细胞介素10表达的影响

李欣; 陈辉; 李红艺; 王瑞丽; 孙淑杰; 任江波; 马红

脂联素对大鼠原代Kupffer细胞白细胞介素10表达的影响

首都医科大学附属北京友谊医院肝病中心

基金项目:

中国肝炎防治基金会王宝恩肝纤维化研究基金资助项目(20070027)；

详细信息

中图分类号: R575
计量
- 文章访问数: 3274
- HTML全文浏览量: 6
- PDF下载量: 597
- 被引次数: 0
出版历程
- 出版日期: 2013-04-20

Effects of adiponectin on IL-10 expression in rat primary Kupffer cells

Research funding:

摘要

摘要:
目的本研究观察外源性脂联素对体外培养的大鼠原代Kupffer细胞白细胞介素（IL）-10表达的调节作用,以阐明脂联素的抗炎作用机制。方法通过两步胶原酶灌流、密度梯度离心和2 h选择性贴壁方法分离纯化大鼠肝脏Kupffer细胞。酶联免疫吸附试验方法检测脂联素作用下原代Kupffer细胞上清液中IL-10的表达水平。荧光定量PCR法检测脂联素作用下原代Kupffer细胞IL-10 mRNA表达。结果 0.5、1.0、1.5μg/ml脂联素处理Kupffer细胞4、8、12、16 h,与对照组相比,同一浓度的脂联素随着培养时间的延长,Kupffer细胞IL-10分泌逐渐增加;而同一作用时间下,随着脂联素浓度的增加,Kupffer细胞IL-10分泌量并没有逐渐增加。1.0和1.5μg/ml脂联素作用Kupffer细胞16 h,IL-10基因表达量与对照组相比显著增加,各处理组间无明显差异。结论脂联素可上调大鼠原代Kupffer细胞IL-10的表达,这可能是其抗炎作用机制之一。
- 脂联素 /
- 枯否细胞 /
- 白细胞介素10
Abstract:
Objective To investigate whether adiponectin exposure can induce expression of the anti-inflammatory cytokine, interleukin-10 (IL-10) , in Kupffer cells, a major source of inflammatory cytokines in liver.Methods Primary Kupffer cells were isolated from male Wistar rats by two-step collagenase perfusion and Percoll density gradient centrifugation, purified by selective adherence in culture, and confirmed by immunofluorescence detection of ED2 expression.The Kupffer cells were treated with 0.5, 1.0 or 1.5 μg/ml of adiponectin.Untreated cells served as controls.At 4, 8, 12 and 16 h of exposure, IL-10 protein levels in supernatant were detected by enzyme-linked immunosorbent assay, and IL-10 mRNA levels were measured by real-time reverse transcription PCR.Results None of the concentrations of adiponectin stimulated IL-10 secretion to a level significantly higher than levels detected in control supernatants when exposed to the cells for 4, 8 or 12 h.However, after 16 hours of exposure, the adiponectin treatments did stimulate the IL-10 secretion to significantly higher levels than that in the control group (all P<0.05) .In addition, after 16 h of exposure, the 1.0 and 1.5 g/ml treatment groups showed significantly higher IL-10 mRNA levels than the control group (both P<0.05) .However, the levels of mRNA stimulation were not significantly different between the two treatment groups.Conclusion Adiponectin up-regulates IL-10 expression in primary cultured rat Kupffer cells, which may act as an anti-inflammatory mechanism of adiponectin.
- adiponectin /
- Kupffer cells /
- interleukin-10

HTML全文

参考文献(14)

[1] Kiso K, Ueno S, Fukuda M, et al. The role of Kupffer cells in carbon tetrachloride intoxication in mice[J]. Biol Pharm Bull, 2012, 35 (6) : 980-983.

[2]Das SK, Balakrishnan V.Role of cytokines in the pathogene-sis of non-alcoholic fatty liver disease[J].Ind J Clin Bio-chem, 2011, 26 (2) :202-209.

[3] An L, Wang X, Cederbaum AI. Cytokines in alcoholic liver disease[J]. Arch Toxicol, 2012, 86 (9) : 1337-1348.

[4]O’Shea JJ, Murray PJ.Cytokine signaling modules in inflam-matory responses[J].Immunity, 2008, 28 (4) :477-487.

[5]Friedman SL.Molecular regulation of hepatic fibrosis, an in-tegrated cellular response to tissue injury[J].J Biol Chem, 2000, 275 (4) :2247-2250.

[6]You M, Considine RV, Leone TC, et al.Role of adiponectinin the protective action of dietary saturated fat against alco-holic fatty liver in mice[J].Hepatology, 2005, 42 (3) :568-577.

[7]Esfandiari F, You M, Villanueva JA, et al.S-adenosylme-thionine attenuates hepatic lipid synthesis in micropigs fedethanol with a folate-deficient diet[J].Alcohol Clin ExpRes, 2007, 31 (7) :1231-1239.

[8]Song Z, Zhou Z, Deaciuc I, et al.Inhibition of adiponectin pro-duction by homocysteine:a potential mechanism for alcoholicliver disease[J].Hepatology, 2008, 47 (3) :867-879.

[9] Ajmo JM, Liang X, Rogers CQ, et al. Resveratrol alleviates alcoholic fatty liver in mice[J]. Am J Physiol Gastrointest Liver Physiol, 2008, 295 (4) : G833-842.

[10]Xu A, Wang Y, Keshaw H, et al.The fat-derived hormone ad-iponectin alleviates alcoholic and nonalcoholic fatty liver diseasesin mice[J].J Clin Invest, 2003, 112 (1) :91-100.

[11] Thakur V, Pritchard MT, McMullen MR, et al. Adiponectin normalizes LPS-stimulated TNF-alpha production by rat Kupffer cells after chronic ethanol feeding[J]. Am J Physiol Gastrointest Liver Physiol, 2006, 290 (5) : 998-1007.

[12]Park PH, McMullen MR, Huang H, et al.Short-term treat-ment of RAW264.7 macrophages with adiponectin increasestumor necrosis factor-alpha (TNF-alpha) expression viaERK1/2 activation and Egr-1 expression:role of TNF-al-pha in adiponectin-stimulated interleukin-10 production[J].J Biol Chem, 2007, 282 (30) :21695-21703.

[13]Mandal P, Pritchard MT, Nagy LE.Anti-inflammatory path-ways and alcoholic liver disease:Role of an adiponectin/inter-leukin-10/heme oxygenase-1 pathway[J].World J Gastro-enterol, 2010, 16 (11) :1330-1336.

[14]Mandal P, Park PH, McMullen MR, et al.The anti-inflam-matory effects of adiponectin are mediated via a heme oxy-genase-1-dependent pathway in rat Kupffer cells[J].Hepatology, 2010, 51 (4) :1420-1429.

施引文献

资源附件(0)

访问统计