| 卢杰,付鑫,张继红,等.ApoE基因敲除小鼠主动脉肿瘤坏死因子-α/p38丝裂原活化蛋白激酶/血清核因子-κB/视黄醇结合蛋白信号通路变化研究[J].安徽医药,2018,22(7):1278-1281. |
| ApoE基因敲除小鼠主动脉肿瘤坏死因子-α/p38丝裂原活化蛋白激酶/血清核因子-κB/视黄醇结合蛋白信号通路变化研究 |
| Changes of TNF-α/p38MAPK/NF-κB/RBP4 signaling pathway in aorta of ApoE-/- mice |
| 投稿时间：2016-12-24 |
| DOI： |
| 中文关键词: 动脉粥样硬化 肿瘤坏死因子-α p38丝裂原活化蛋白激酶类 NF-κB 视黄醇结合蛋白质类 小鼠 |
| 英文关键词: Atherosclerosis Tumor necrosis factor-alpha p38 mitogen-activated protein kinases NF-kappa B Retinol-binding proteins Mice |
| 基金项目:沈阳市科技局计划项目(F15-139-9-03) ApoE基因敲除小鼠主动脉肿瘤坏死因子-α/p38丝裂原活化蛋白激酶/血清核因子-κB/视黄醇结合蛋白信号通路变化研究卢杰,付鑫,张继红,陈红光,高俊甲,范军,张颖 (沈阳医学院附属第二医院心血管内科,辽宁 沈阳 110036) |
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| 中文摘要: |
| 目的 通过观察ApoE基因敲除动脉粥样硬化小鼠主动脉肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)/p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinases ,p38MAPK)/血清核因子-κ B(Nuclear Factor κB,NF-κ B)/视黄醇结合蛋白(Retinol Binding Protein-4,RBP4)信号通路变化,探讨RBP4在动脉粥样硬化的炎症反应过程的作用及发生机制。方法 10只ApoE-/-小鼠给予高脂饲料喂养8周,进行动脉粥样硬化模型复制,空白对照组选取具有相同遗传背景的同龄野生型C57BL/6J小鼠10只。饲养8周后,分离两组小鼠血清,分离主动脉,分别保存于4%多聚甲醛和-80 ℃冰箱。Elisa法检测小鼠血清TNF-α、RBP4水平变化,全自动生化分析仪检测血清中甘油三酯(Triglyceride,TG)、低密度脂蛋白(Low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白(High density lipoprotein cholesterol,HDL-C)、血清总胆固醇(Total cholesterol,TC)水平,HE染色法观察两组小鼠主动脉组织形态学变化,蛋白质印迹法和RT-PCR法检测主动脉TNF-α、p38MAPK、NF-κB、RBP4蛋白和基因表达情况。结果 血脂水平检测发现,与空白对照组相比,模型组小鼠血清中TG、TC、LDL-C水平显著升高(P<0.01),HDL-C水平显著降低(P<0.01)；HE 染色观察发现,较空白对照组小鼠,模型组实验小鼠主动脉管腔内可见较大的AS斑块形成。蛋白质印迹法(western blot)结果显示,与空白对照组比较,模型组小鼠TNF-α[(0.93±0.05)mg·L-1,t=-9.079,P<0.001)]、NF-κB[(0.82±0.03) mg·L-1,t=-9.718,P<0.001]水平明显升高,p38MAPK、RBP4水平显著升高(P<0.01)。PCR结果显示,模型组小鼠TNF-α(0.78±0.03 mg·L-1)和RBP4(0.39±0.02 mg·L-1)水平较空白对照组明显升高(P<0.05,P<0.01)。结论 TNF-α/p38MAPK/ NF-κB / RBP4信号通路可能参与了动脉粥样硬化的形成及炎症反应过程。 |
| 英文摘要: |
| Objective To investigate the role and mechanism of RBP4 in the inflammatory reaction of atherosclerosis by observing the changes of TNF-α/p38MAPK/ NF-κB/RBP4 signaling pathway in aorta of ApoE-/- atherosclerosis mice. Methods 10 ApoE-/- mice were fed with high fat diet for 8 weeks, to replicate the model of atherosclerosis. Ten C57BL/6J mice with same age and genetic background were composed in blank control group. 8 weeks later, the serum and aorta of mices was separated. Elisa method was used to detect the changes of serum TNF-αand RBP4 levels in mice. The content of TG, TC, HDL-C, LDL-C in serum were detected by automatic biochemical analyzer. HE staining method was used to observe the morphological changes of the aorta in two groups of mice. Blot Western and RT-PCR method were adopted to detect the expression of TNF-α, p38MAPK, NF-κB and RBP4 in aorta. Results Compared with blank control group, model group has significantly increased in TC, TG, LDL-C levels (P<0.01), decreased in HDL-C level (P<0.01). HE staining showed that, compared with the blank control group, the model group, the larger the AS plaque formation was observed in the aorta lumen of the model group. Blot Western results show that compared with the blank control group, in model group, the levels of TNF-α[(0.93±0.05) mg·L-1, t=-9.079, P<0.001], NF-κB[(0.82±0.03) mg·L-1, t=-9.718, P<0.001]were significantly increased (P<0.05). RBP4 and p38MAPK levels were significantly elevated (P<0.01). PCR results showed that, compared with the blank control group, the levels of TNF-α (0.78±0.03) mg·L-1 and RBP4 (0.39±0.02) mg·L-1 in model group were significantly increased (P<0.05,P<0.01). Conclusion TNF-α/p38MAPK/ NF-κB/RBP4 signaling pathway may be involved in the formation of atherosclerosis and inflammatory reaction process. |
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