丝裂原激活蛋白激酶激酶激酶激酶4在动脉粥样硬化易损斑块形成中的调节作用  

Regulatory role of Map4k4 protein expression in atherosclerotic vulnerable plaque formation

作　　者：陶晓芳 王雅洁 王文斌 费年华 TAO Xiaofang;WANG Yajie;WANG Wenbin;FEI Nianhua(Department of Geriatrics,Xianning Central Hospital(the First Affiliated Hospital of Hubei University of Science and Technology),Xianning 437000,China)

机构地区：[1]咸宁市中心医院(湖北科技学院附属第一医院)老年病科,湖北咸宁437000 [2]咸宁市中心医院(湖北科技学院附属第一医院)甲状腺乳腺外科,湖北咸宁437000

出　　处：《实用医学杂志》2022年第11期1346-1352,共7页The Journal of Practical Medicine

基　　金：湖北省科技厅科研项目(编号:WJ2019M096)。

摘　　要：目的 探讨丝裂原激活蛋白激酶激酶激酶激酶4(Map4k4)在血管紧张素Ⅱ(Ang Ⅱ)诱导的动脉粥样硬化(AS)易损斑块形成中的调节作用。方法 ApoE^(-/-)小鼠随机分为4组(每组8只):Control组、Ang Ⅱ组、Ang Ⅱ+shMap4k4组和Ang Ⅱ+Map4k4组。采用油红O染色(脂质)、Masson染色(胶原)、α-肌动蛋白免疫荧光染色(平滑肌细胞)和F4/80染色(巨噬细胞)检测主动脉窦斑块稳定性。通过免疫荧光和Western blot分析Map4k4表达。体外构建Map4k4敲低或过表达巨噬细胞,用于确认Map4k4在Ang Ⅱ诱导的细胞凋亡中的作用。结果 Ang Ⅱ促进了ApoE^(-/-)小鼠AS斑块易损性和巨噬细胞凋亡(P <0.05),伴随着Map4k4表达的上调(P <0.05)。与Ang Ⅱ组小鼠相比,Ang Ⅱ+shMap4k4组小鼠的AS斑块易损性[(1.22±0.06)vs.(0.49±0.04)]和巨噬细胞凋亡[(8.42±0.40)vs.(3.49±0.31)]明显减弱(P <0.05),而Ang Ⅱ+Map4k4组小鼠上述指标明显增强[(1.22±0.06)vs.(1.45±0.05),(8.42±0.40)vs.(14.11±0.78),P <0.05]。体外实验显示,Map4k4敲低有效减弱了Ang Ⅱ诱导的c-Caspase3、Bax蛋白表达增加(P <0.05),并增加了Bcl-2蛋白表达(P <0.05)。而Map4k4过表达进一步增强了Ang Ⅱ的诱导作用(P <0.05)。结论Ang Ⅱ通过上调Map4k4表达促进巨噬细胞凋亡,并进一步参与AS中易损斑块的形成。Objective To explore the regulatory role of mitogen-activated protein kinase kinase kinase kinase 4(Map4k4)in the formation of vulnerable plaques in atherosclerosis(AS)induced by angiotensin Ⅱ(Ang Ⅱ).Methods Thirty-two ApoE^(-/-)mice were randomly divided into four groups(8 in each group):control,Ang Ⅱ,Ang Ⅱ+shMap4k4 and Ang Ⅱ+Map4k4. Oil red O staining(lipid),Masson staining(collagen),α-actin immunofluorescence staining(smooth muscle cells)and F4/80 staining(macrophages)were used to detect the stability of aortic sinus plaque. The expression of Map4k4 was analyzed by immunofluorescence and Western blot. Map4k4 knockdown or overexpression macrophages were constructed in vitro to confirm the role of Map4k4 in Ang Ⅱ-induced apoptosis. Results The atherosclerotic plaque vulnerability and macrophage apoptosis were promoted in ApoE^(-/-)mice by Ang Ⅱ(P < 0.05),accompanied by the upregulation of Map4k4 expression. Compared with the Ang Ⅱ group(P < 0.05),the AS plaque vulnerability[(1.22 ± 0.06)vs.(0.49 ± 0.04)]and macrophage apoptosis[(8.42 ± 0.40)vs.(3.49 ± 0.31)]of the Ang Ⅱ+shMap4k4 group were significantly reduced(P < 0.05),while the above indicators of the Ang Ⅱ+Map4k4 group were significantly enhanced[(1.22 ± 0.06)vs.(1.45 ± 0.05);(8.42 ± 0.40)vs.(14.11 ± 0.78),P < 0.05]. In vitro experiments showed that Map4k4 knockdown effectively attenuated the increased expression of c-Caspase3 and Bax protein induced by Ang Ⅱ(P < 0.05),and enhanced the expression of Bcl-2 protein(P < 0.05). The overexpression of Map4k4 further enhanced the induction of Ang Ⅱ(P < 0.05). Conclusion Ang Ⅱ via upregulating Map4k4 expression promotes macrophage apoptosis so as to promote vulnerable plaque formation in AS.

关 键 词：丝裂原激活蛋白激酶激酶激酶激酶4 血管紧张素Ⅱ 动脉粥样硬化 易损斑块 巨噬细胞 

分 类 号：R541.4[医药卫生—心血管疾病]