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肥胖对脓毒症小鼠模型的肝脏损害作用及其对炎症反应的影响
Effects of obesity on liver injury and inflammatory response in septic mouse model
- 重庆医科大学学报 2017年42卷第12期 页码：1620-1626
- 作者机构：
重庆市急救医疗中心肝胆外科
- 作者简介：
- 基金信息：
DOI：10.13406/j.cnki.cyxb.001433
中图分类号： R-332;R459.7纸质出版日期：2017，
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周玉庆, 陈翔. 肥胖对脓毒症小鼠模型的肝脏损害作用及其对炎症反应的影响[J]. 重庆医科大学学报, 2017,42(12):1620-1626.
Zhou Yuqing, Chen Xiang. Effects of obesity on liver injury and inflammatory response in septic mouse model[J]. Journal of Chongqing Medical University, 2017,42(12):1620-1626.
周玉庆, 陈翔. 肥胖对脓毒症小鼠模型的肝脏损害作用及其对炎症反应的影响[J]. 重庆医科大学学报, 2017,42(12):1620-1626. DOI： 10.13406/j.cnki.cyxb.001433.
Zhou Yuqing, Chen Xiang. Effects of obesity on liver injury and inflammatory response in septic mouse model[J]. Journal of Chongqing Medical University, 2017,42(12):1620-1626. DOI： 10.13406/j.cnki.cyxb.001433.
摘要
目的:探讨肥胖对脓毒症小鼠肝损害和炎症反应的影响。方法:6周龄C57BL/6J野生型雄性小鼠80只随机分为高脂饮食组(HFD组
40只)和普通饮食组(ND组
40只)
饲养6周后称重
测葡萄糖耐量
比色法检测肝脏甘油三酯(triglyceride
TG)水平。采用盲肠结扎穿刺(cecal ligation-peferation
CLP)法构建脓毒症模型
观察小鼠生存率
取血浆和肝脏组织
HE染色观察小鼠肝脏组织
采用酶联免疫法(enzyme-linked immunosorbent assay
ELISA)检测谷丙转氨酶(alanine aminotransferase
ALT)、瘦素(leptin)、脂联素(adiponectin)、肿瘤坏死因子-α(tumor necrosis factor
TNF-α)、白介素-6(interleukin-6
IL-6)及IL-10的表达
RT-PCR检测活化蛋白(activator protein-1
AP-1)mRNA的表达。结果:HFD组小鼠体质量明显重于ND组小鼠[(34.867±1.465)vs.(25.678±1.761)
t=-9.827
P=0.000]
负荷实验示:HFD组与ND组相比
血糖水平更高(F=200.921
P=0.000)
肝脏甘油三酯水平更高(t=31.793
P=0.000)。肥胖小鼠相对于普通饮食小鼠脓毒症时生存率更低
生存时间更短(P=0.037)。经CLP处理后
肥胖小鼠血浆瘦素水平仍明显高于非肥胖小鼠[(27.64±5.16)vs.(7.12±2.91)ng/mL
F=95.141
P=0.000]
而脂联素在肥胖小鼠脓毒症时表达降低[6 h ND组vs.HFD组:(8 445.344±3 245.632)vs.(2 434.532±2.012)
t=2.660
P=0.000]。脓毒症时小鼠血浆TNF-α、IL-6、IL-10表达均增加[TNF-α:(2.017±1.071)vs.(170.698±37.705)
t=8.137
P=0.001;(2.760±0.853)vs.(122.080±19.520)
t=13.553
P=0.000;IL-6:(19.955±3.500)vs.(404.752±50.701)
t=18.070
P=0.000;(15.975±4.022)vs.(533.064±104.866)
t=11.032
P=0.000;IL-10:(79.810±7.042)vs.(277.156±124.213)
t=4.335
P=0.012;(120.678±7.172)vs.(214.111±85.322)
t=3.925
P=0.020]
但肥胖小鼠相对非肥胖小鼠TNF-α和IL-10表达降低[TNF-α:(170.698±37.705)vs.(122.080±19.520)
t=0.981
P=0.356;IL-10:(277.156±124.213)vs.(214.111±85.322)
t=3.216
P=0.031]。血浆ALT的水平在肥胖的脓毒症小鼠中明显高于非肥胖的脓毒症小鼠[(132.774±45.662)vs.(236.633±42.782)
t=2.831
P=0.045]。CLP刺激后
AP-1在非肥胖小鼠活性增加
但在肥胖小鼠中活性增加更显著[ND 0 h vs.6 h
(1.069±0.404)vs.(1.653±0.184)
F=32.886
P=0.000;6 h ND vs.HFD:(1.653±0.184)vs.(5.021±0.623)
F=29.189
P=0.000]。结论:高脂饮食诱导的肥胖小鼠在脓毒症时生存率低
肝脏损伤较重
炎症反应降低
这可能与AP-1活性增加有关。
Abstract
Objective:To explore the role of obesity in sepsis-induced liver injury and inflammation.Methods:The experimental groups consisted of 80 male C57 BL/6 mice at 6 weeks of age.Animals were randomized to HFD（n=40）or ND（n=40）for 6 weeks
then they were weighed.Glucose tolerance tests were measured.The liver triglyceride levels were determined by colorimetric assay kit.Poly microbial sepsis was induced by cecal ligation-peferation（CLP） and mice were monitored for survival.Plasma and liver tissues were collected.HE staining was used to observe the liver tissue of mice.The levels of alanine aminotransferase（ALT）
leptin
adiponectin
TNF-α
IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay（ELISA）.RT-PCR was used to detect the expression of activator protein-1 expression.Results:The body weight of HFD group was significantly higher than that of ND group[（34.867±1.465）vs.（25.678±1.761）
t=-9.827
P=0.000].Glucose tolerance test:compared with the ND group
the HFD group had higher blood glucose levels and higher hepatic triglyceride levels（F=200.921
P=0.000;t=31.793
P=0.000）.Obese mice had a lower survival rate and a shorter survival time（P=0.037）than ND mice in sepsis.After CLP treatment
the levels of plasma leptin in obese mice were still significantly higher than those in non-obese mice [（27.64 ±5.16） ng/mL vs.（7.12 ±2.91） ng/mL
F =95.141
P=0.000]
while adiponectin expressions were decreased in obese mice[6 h ND group vs.HFD group:（8 445.344±3 245.632）vs.（2 434.532±2.012）
t=2.660
P=0.000].The expressions of TNF-α
IL-6 and IL-10 were increased in septic mice[TNF-α:（2.017±1.071）vs.（170.698±37.705）
t=8.137
P=0.001;（2.760±0.853）vs.（122.080±19.520）
t=13.553
P=0.000;IL-6:（19.955±3.500）vs.（404.752±50.701）
t=18.070
P=0.000;（15.975±4.022）vs.（533.064±104.866）
t=11.032
P=0.000;IL-10:（79.810±7.042）vs.（277.156±124.213）
t=4.335
P=0.012;（120.678±7.172）vs.（214.111±85.322）
t=3.925
P=0.020]
but the expression of TNF-α and IL-10 in obese mice was lower than that in non-obese mice[TNF-α:（170.698±37.705）vs.（122.080±19.520）
t=0.981
P=0.356;IL-10:（277.156±124.213）vs.（214.111±85.322）
t=3.216
P=0.031].Plasma ALT levels were significantly higher in obese sepsis mice than in non-obese sepsis mice[（132.774 ±45.662）vs.（236.633±42.782）
t=2.831
P=0.045].Although AP-1 activity increased in mice without obesity at 6 h after CLP
AP-1 activity was significantly higher in mice with obesity than in mice without obesity after CLP[ND 0 h vs.6 h:（1.069±0.404）vs.（1.653±0.184）
F=32.886
P=0.000;6 h ND vs.HFD:（1.653±0.184）vs.（5.021 ±0.623）
F=29.189
P=0.000].Conclusion:High fat diet induced obese mice in sepsis have low survival rate
severe liver injury
decreased inflammatory response
which may be related to increased activity of AP-1.
关键词
脓毒症肥胖肝损害炎症反应
Keywords
sepsisobesityliver injuryinflammatory response
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