模拟失重下miR-212-3p/SIRT1通路调控微血管内皮细胞增殖  

MiR-212-3p/SIRT1 pathway regulates microvascular endothelial cell proliferation under simulated microgravity

作　　者：徐丽群 张晓雁 张丽君 孙权 薛桐 胡泽兵 曹新生 石菲 张舒 XU Liqun;ZHANG Xiaoyan;ZHANG Lijun;SUN Quan;XUE Tong;HU Zebing;CAO Xinsheng;SHI Fei;ZHANG Shu(Department of Aerospace Biodynamics,School of Aerospace Medicine,Air Force Medical University,Ministry-of-Education Key Laboratory of Aerospace Medicine,Xi'an 710032,China)

机构地区：[1]空军军医大学航空航天医学系生物动力学教研室,航空航天医学教育部重点实验室,陕西西安710032

出　　处：《空军军医大学学报》2022年第1期6-12,共7页Journal of Air Force Medical University

基　　金：国家自然科学基金(31971171)。

摘　　要：目的 探究模拟失重下miR-212-3p/SIRT1信号通路对微血管内皮细胞增殖的影响,为失重导致的心血管功能失调的在体靶向干预寻求治疗靶点。方法 利用2D回转器对小鼠脑微血管内皮细胞进行模拟失重处理,探究miR-212-3p及沉默信息调节因子2相关酶I(SIRT1)在模拟失重下的表达变化;向bEnd.3细胞中转染mimic-212-3p、inhibitor-212-3p、pcDNA3.1-SIRT1及相应的阴性对照进行功能验证或挽救实验;采用qRT-PCR检测miR-212-3p及SIRT1 mRNA表达变化;采用Western blotting检测PCNA、SIRT1蛋白的表达变化;采用CCK-8及5-乙炔基-2′-脱氧尿苷(EdU)检测微血管内皮细胞增殖能力。结果 模拟失重下,bEnd.3细胞中miR-212-3p表达升高、而SIRT1表达下降(P<0.01);转染mimic-212-3p、inhibitor-212-3p后,Western blotting、CCK-8及EdU检测均表明过表达miR-212-3p能够显著抑制微血管内皮细胞增殖,抑制miR-212-3p能够促进微血管内皮细胞增殖(P<0.01);此外,沉默miR-212-3p可以缓解模拟失重导致的微血管内皮细胞增殖抑制,表现为模拟失重下转染inhibitor-212-3p后PCNA蛋白表达增加,细胞增殖率升高(P<0.01);mimic-212-3p与pcDNA3.1-SIRT1共转染实验表明miR-212-3p通过抑制SIRT1从而负调控微血管内皮细胞增殖(P<0.01)。结论 模拟失重促进miR-212-3p表达,而抑制SIRT1表达;miR-212-3p通过直接负调控SIRT1从而抑制微血管内皮细胞增殖,提示抑制miR-212-3p/SIRT1通路可以部分降低模拟失重对微血管内皮细胞的增殖抑制,miR-212-3p可以作为潜在的干预靶点。Objective To explore the effect of miR-212-3 p/SIRT1 signaling pathway on the proliferation of microvascular endothelial cells under simulated microgravity, and provide a therapeutic target for intervention in vivo of cardiovascular dysfunction induced by microgravity. Methods Mouse cerebral microvascular endothelial cells were cultured under simulated microgravity by 2 D clinostat to observe the changes in the expression of miR-212-3 p and silent information regulator 1(SIRT1)under simulated microgravity. Mimic-212-3 p, inhibitor-212-3 p, pcDNA3.1-SIRT1 and the corresponding negative controls were transfected into bEnd.3 cells for functional verification or rescue experiments. The qRT-PCR was used to detect the expression of miR-212-3 p and SIRT1. The protein expression of PCNA and SIRT1 was detected by Western blotting. CCK-8 and EdU assays were used to detect the proliferation ability of microvascular endothelial cells. Results The expression of miR-212-3 p increased, while the expression of SIRT1 decreased in bEnd.3 cells under simulated microgravity(P<0.01);after transfection with mimic-212-3 p and inhibitor-212-3 p, Western blotting, CCK-8 and EdU assays showed that over-expression of miR-212-3 p significantly inhibited the proliferation of microvascular endothelial cells(P<0.01);in addition, silencing miR-212-3 p alleviated the suppression of microvascular endothelial cell proliferation induced by simulated microgravity, manifested as increased expression of PNCA and cell proliferation rate after transfection with inhibitor-212-3 p under simulated microgravty(P<0.01). After mimic-212-3 p and pcDNA3.1-SIRT1 were co-transfected into bEnd.3 cells, miR-212-3 p negatively regulated the proliferation of the microvascular endothelial cells by inhibiting SIRT1(P<0.01). Conclusion Simulated microgravity can promote the expression of miR-212-3 p and inhibit the expression of SIRT1. The miR-212-3 p can inhibit the proliferation of microvascular endothelial cells by regulating SIRT1, suggesting that inhibition of miR-

关 键 词：miR-212-3p SIRT1 模拟失重 细胞增殖 小鼠脑微血管内皮细胞 

分 类 号：R285[医药卫生—中药学]