摘要
目的:探讨环磷酸鸟苷-腺苷合成酶-干扰素基因刺激因子(cGAS-STING)信号通路对心肌缺血再灌注损伤(MIRI)动物模型中巨噬细胞极化的调控作用。方法:40只健康雄性C57BL/6小鼠,随机均分为对照组、假手术组、模型组和RU.521(cGAS抑制剂)组。模型组和RU.521组构建MIRI模型,假手术组仅开胸但不结扎冠脉,对照组无手术操作。RU.521组给予5 mg·kg^(-1)·d^(-1)的RU.521腹腔注射,其余3组注射等体积0.9%氯化钠注射液,连续7 d。实验结束后,收集心肌组织与血清样本,采用蛋白免疫印迹(WB)法检测cGAS、STING蛋白表达,流式细胞术(FCM)测定M1、M2型巨噬细胞占比,酶联免疫吸附(ELISA)法检测IL-6、TNF-α、IL-10、TGF-β含量。结果:与对照组和假手术组相比,模型组小鼠的cGAS、STING蛋白表达水平,M1型巨噬细胞占比以及IL-6、TNF-α含量均显著升高,M2型巨噬细胞占比及IL-10、TGF-β水平均显著降低(P<0.001);与模型组比较,RU.521组小鼠的GAS、STING蛋白表达水平,M1型巨噬细胞占比以及IL-6、TNF-α含量均显著下降,M2型巨噬细胞占比及IL-10、TGF-β水平均显著升高(P<0.01)。结论:缺血再灌注损伤触发cGAS-STING信号通路的激活,可促进巨噬细胞向M1型极化,并增强炎症反应,而抑制该通路则有助于巨噬细胞向M2型极化,从而限制过度激活的炎症反应。
Objective:To investigate the regulation of cyclic guanosine monophosphate-adenosine monophosphate synthasestimulator of interferon genes(cGAS-STING)signal pathway on macrophage polarization in an animal model of myocardial isch-emia-reperfusion injury(MIRI).Methods:A total of 40 healthy male C57BL/6 mice were randomly divided into the control group,the sham group,the model group,and the RU.521(cGAS inhibitor)group,with 10 mice in each group.The model of MIRI was constructed in the model group and RU.521 group,the sham group had only thoracotomy but no coronary ligation,and the control group had no surgical procedure.The RU.521 group was intraperitoneally injected at a dose of 5 mg/kg/d,and the remaining three groups were injected with an equal volume of 0.9%sodium chloride injection for 7 d.At the end of the experiment,myocardial tissue and serum samples were collected,the expression of cGAS and STING proteins was detected by Western blot(WB),the proportion of M1 and M2 macrophages was determined by flow cytometry,and the contents of IL-6,TNF-α,IL-10 and TGF-βwere detected by enzyme-linked immunosorbent assay.Results:Compared to the control group and sham group,the expression levels of cGAS and STING proteins,the proportion of M1 macrophages,and the contents of IL-6 and TNF-αwere significantly increased,but the proportion of M2 macrophages and the levels of IL-10 and TGF-βwere significantly decreased in model group(P<0.001);Compared to the model group,the levels of GAS and STING protein expression,the proportion of M1 macrophages,and the contents of IL-6 and TNF-αwere significantly decreased,but the proportion of M2 macrophages and the levels of IL-10 and TGF-βwere significantly increased in the RU.521 group(P<0.01).Conclusion:Ischemia-reperfusion trig-gers activation of the cGAS-STING signaling pathway to promote the polarization of macrophages towards type M1 and enhance the inflammatory response.However,inhibition of this signaling pathway contributes to the polarization of macrophages towards type M2,which can limit the overactivated inflammatory response.
作者
张焕鑫
沈祥礼
葛振嵘
王潇
宋涛
姜述斌
ZHANG Huanxin;SHEN Xiangli;GE Zhenrong;WANG Xiao;SONG Tao;JIANG Shubin(The Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,China)
出处
《海南医科大学学报》
北大核心
2025年第3期178-184,共7页
Journal of Hainan Medical University
基金
新疆维吾尔自治区自然科学基金项目(2023D01C138)。
