Interleukin-10 inhibits the in vivo and in vitro adverse effects of TNF-alpha on the endothelium of murine aorta.
Interleukin-10 inhibits the in vivo and in vitro adverse effects of TNF-alpha on the endothelium of murine aorta.
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DOI：
10.1152/ajpheart.00763.2009
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发表时间：
2010-10
期刊：
影响因子：
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通讯作者：
Saiprasad M. Zemse;C. Chiao;R. Hilgers;R. Webb
中科院分区：
文献类型：
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作者：
Saiprasad M. Zemse;C. Chiao;R. Hilgers;R. Webb
TNF-α是一种促炎细胞因子，是母体内皮功能障碍导致子痫前期的重要介质。在本研究中，我们检测了IL-10是否对TNF-α-诱导的小鼠主动脉内皮功能障碍有保护作用。在体外实验中，将C57BL/6雌性小鼠的主动脉环在Dulbecco改良Eagle培养基中孵育，在37℃条件下，分别存在载体（蒸馏H(2)O）、TNF-α (4 nmol/l)或重组小鼠IL-10 (300 ng/ml)，或同时存在TNF-α和IL-10 22 H。在体内实验中，C57BL6/IL-10基因敲除雌性小鼠用生理盐水或TNF-α （220 ng·kg（-1）·day(-1)）治疗14天。从体外和体内实验中分离主动脉环，安装在钢丝肌图（丹麦Myotech）上，拉伸至5 mN的张力。在苯肾上腺素（10 μmol/l）诱导的收缩过程中，构建乙酰胆碱（ACh, 0.001 ~ 10 μmol/l）对内皮依赖性松弛的累积浓度响应曲线。结果，与未处理的环相比，将主动脉环暴露于TNF-α中过夜导致对ACh的最大放松反应（E(max)）明显减弱（分别为22±4比82±3%）。与TNF-α处理的C57BL/6小鼠相比，IL-10敲除小鼠与TNF-α处理的ACh反应明显受损（E(max)）（分别为51±3比72±3%）。Western blot分析结果显示，体外和体内实验中，TNF-α可降低内皮细胞一氧化氮合酶（eNOS）的表达，而IL-10可恢复eNOS的表达。综上所述，抗炎细胞因子IL-10通过保护eNOS的表达来预防TNF-α引起的内皮依赖性血管舒张损伤。
TNF-α is a proinflammatory cytokine and is an important mediator of maternal endothelial dysfunction leading to preeclampsia. In this study, we tested whether IL-10 protects against TNF-α-induced endothelial dysfunction in murine aorta. In in vitro experiments, aortic rings of C57BL/6 female mice were incubated in Dulbecco's modified Eagle's medium in the presence of either vehicle (distilled H(2)O), TNF-α (4 nmol/l), or recombinant mouse IL-10 (300 ng/ml) or in the presence of both TNF-α and IL-10 for 22 h at 37°C. In in vivo experiments C57BL6/IL-10 knockout female mice were treated with saline or TNF-α (220 ng·kg(-1)·day(-1)) for 14 days. Aortic rings were isolated from in vitro and in vivo experiments and mounted in a wire myograph (Danish Myotech) and stretched to a tension of 5 mN. Endothelium-dependent relaxation was assessed by constructing cumulative concentration-response curves to acetylcholine (ACh, 0.001-10 μmol/l) during phenylephrine (10 μmol/l)-induced contraction. As a result, overnight exposure of aortic rings to TNF-α resulted in significant blunted maximal relaxing responses (E(max)) to ACh compared with untreated rings (22 ± 4 vs. 82 ± 3%, respectively). IL-10 knockout mice treated with TNF-α showed significant impairment in ACh responses (E(max)) compared with C57BL/6 mice treated with TNF-α (51 ± 3 vs. 72 ± 3%, respectively). Western blot analysis showed that endothelial nitric oxide synthase (eNOS) expression was reduced by TNF-α in in vitro and in vivo experiments, whereas IL-10 restored the eNOS expression. In conclusion, the anti-inflammatory cytokine IL-10 prevents impairment in endothelium-dependent vasorelaxation caused by TNF-α by protecting eNOS expression.