骨髓来源单核巨噬细胞对高血压小鼠心脏损伤和重构的影响  

Effect of bone marrow derived macrophages on hypertension induced cardiac injury and remodeling in mice

作　　者：李玉琳[1] 张聪聪[1] 吴依娜[1] 王春筱[1] 阿希[1] 王吉静[1] 王月丽[1] 王绿娅[1] 杜杰[1] 

机构地区：[1]北京市心肺血管疾病研究所,100029

出　　处：《中国心血管杂志》2013年第2期116-120,共5页Chinese Journal of Cardiovascular Medicine

基　　金：国家自然科学基金(81230006;81000069);北京市自然科学基金(7132043;2010B031)~~

摘　　要：目的观察高血压介导骨髓来源单核巨噬细胞在心脏浸润的情况,探讨其对高血压心脏损伤和重构的影响。方法 20只雄性C57BL/6J小鼠随机分为对照组和AngⅡ灌注组,每组10只,采用植入式胶囊渗透压泵分别灌注生理盐水和AngⅡ,采用鼠尾套法监测小鼠动脉血压。于灌注后7 d,采用免疫组织化学、Masson染色和流式细胞技术观察心脏纤维化和心脏中单核巨噬细胞的浸润;采用流式细胞技术和实时荧光定量PCR(rt-PCR)观察循环中单核细胞上CCR2受体表达和心脏中CCR2受体的配体MCP-1表达;16只雄性小鼠随机分为4组,每组4只,采用尾静脉注射脂质体包裹氯膦酸二钠方法清除小鼠体内巨噬细胞,以注射脂质体包裹生理盐水为对照组,分别灌注生理盐水和AngⅡ后观察心脏纤维化。结果与生理盐水灌注对照组相比,AngⅡ灌注后1 d收缩压开始升高,7 d血压显著升高[收缩压:(157.0±13.9)mm Hg比(93.0±11.1)mm Hg,P<0.01],心脏中胶原沉积增加(4.1%±0.7%比0.4%±0.1%,P<0.01),Ⅰ型胶原和肌成纤维细胞的标志物α-SMA的蛋白表达增加。流式检测示心脏中CD45+F4/80+细胞浸润增加;心肌组织半乳糖凝集素2(Mac-2,巨噬细胞标志)和CD68阳性巨噬细胞浸润增加;外周血CD45+CD11B+单核细胞表达CCR2,心脏中MCP-1的mRNA表达增加;小鼠去除巨噬细胞后,AngⅡ灌注7 d的心脏中巨噬细胞浸润减少,心脏纤维化程度减轻。结论高血压促进骨髓来源单核巨噬细胞在心脏的募集,巨噬细胞浸润与心脏纤维化发生相关,去敲除巨噬细胞能够抑制高血压心脏纤维化。Objective It was previously proved that the infihration of bone marrow derived monocyte/macrophage is critical to the inflammatory response. In this study, we aimed to explore the relationship between the infiltration of bone marrow derived monocyte/macrophage and the formation of hypertension-induced cardiac fibrosis. Methods The 20 C57BL/6J mice were infused with a 1500 ng· kg^-1 · min^-1 dose of either 0. 9% saline （saline group,n = 10） or angiotensin Ⅱ （Ang Ⅱ group, n = 10） via an osmotic mini-pump subcutaneously implanted for 7 days. Blood pressure was measured by tail-cuff method to evaluate the effect of Ang Ⅱ infusion. At day 7, AngⅡ or saline infused mice were evaluated by Masson, immunohistochemistry staining and flow cytometry to analyze the cardiac fibrosis and the infiltration of monocyte/macrophage. Flow cytometry and real-time-PCR were used to evaluate the expression of CCR2 on the periphery macrophages and monocyte chemoattractant protein-1 （MCP-1）, the ligand of CCR2 receptor, in the heart tissue. The 16 mice were divided into 4 groups, the intravenous injection of liposome- encapsulated Clodronate was used to clear off the periphery macrophage, the liposome-encapsulated saline was used as control, then mice were infused with either 0. 9% saline or angiotensin Ⅱ to study the fibrosis formation. Results Compared with saline infusion control group, systolic blood pressure began to increase at day 1 and was significantly increased at day 7 after Ang 11 infusion [ （93.0 ± 11.1 ） mm Hg vs. （ 157.0 ± 13. 9）ram Hg, P 〈 0. 01 ]. Collagen deposition was significantly increased （4. 1% - 0. 7% vs. 0. 4% - 0. 1%, P 〈0. 01 ） in the Ang 11 infused heart, and the protein expression levels of collagen I and α-SMA were also increased. Flow cytometry results shown that there were more CD45 ＋ F4/80 ＋ macrophages infiltrated in the Ang Ⅱ infused heart compared with control heart. Immunhistochemistry staining data shown that the galectin 3 （also known as Mac-2） pos

关 键 词：血管紧张素Ⅱ 高血压 心脏重构 巨噬细胞 趋化因子类 

分 类 号：R544.1[医药卫生—心血管疾病]