CD137通过调控血管平滑肌细胞白细胞介素1β分泌参与老年血管慢性炎症的机制研究  

CD137 is involved in aging vascular chronic inflammation by regulating the secretion of IL-1β in vascular smooth muscle cells

作　　者：姜瑜 赵佳 邓冰雁 兰甜宁 卞石惠 赵倩茹 戴芝银[1] 仲威[1] Jiang Yu;Zhao Jia;Deng Bingyan;Lan Tianning;Bian Shihui;Zhao Qianru;Dai Zhiyin;Zhong Wei(Department of Cardiology,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,Jiangsu Province,China)

机构地区：[1]江苏大学附属医院心内科,镇江212001 [2]江苏大学附属人民医院老年科 [3]江苏大学临床医学院

出　　处：《中华老年心脑血管病杂志》2022年第12期1301-1305,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(82000261);江苏省高等学校自然科学研究项目(20KJB320013);江苏省卫生计生委医学科研课题(H201644);江苏大学学生科研项目(20AD0144);镇江市社会发展项目(SH2019071)。

摘　　要：目的 探讨CD137通路参与老年血管慢性炎症的机制。方法 选择C57/B6小鼠40只,CD137小鼠20只,将小鼠分为对照组、衰老组、衰老CD137组,每组20只。通过Luminex液相芯片多因子检测技术检测各组血浆炎性因子表达。通过qPCR技术及免疫荧光技术对各组血管中白细胞介素(IL)-1β进行定量和定位。分别从C57/B6和CD137^(-/-)小鼠主动脉中提取并原代培养血管平滑肌细胞(VSMC)。通过博来霉素体外诱导细胞衰老,采用Western blot、qPCR分别检测各组细胞核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、IL-βmRNA和蛋白表达,采用ELISA检测细胞分泌IL-1β水平。结果 对照组、衰老组、衰老CD137组血浆IL-1β水平分别为(10.86±0.89)pg/ml、(61.40±3.88)pg/ml和(18.20±1.25)pg/ml。衰老组血浆趋化因子配体5、IL-3、IL-13、TNF-α、IL-1β水平较对照组明显升高,衰老CD137^(-/-)组血浆趋化因子配体5、IL-3、IL-13、TNF-α、IL-1β水平较衰老组明显降低(P<0.01)。衰老组血管NLRP3和IL-1βmRNA表达较对照组和衰老CD137组明显升高(3.45±0.62 vs 1.00±0.00,1.57±0.17,P<0.05;5.89±0.82 vs 1.00±0.00,2.64±0.34,P<0.05)。体外实验中,衰老组VSMC中NLRP3和IL-1β蛋白表达较对照组和衰老CD137^(-/-)组明显升高(1.65±0.05 vs 1.00±0.00,0.85±0.05,P<0.05;1.45±0.05 vs 1.00±0.00,1.35±0.05,P<0.05)。衰老组VSMC上清中IL-1β水平较对照组和衰老CD137^(-/-)组明显升高(27.50±4.32 vs 14.20±3.52,16.50±2.72,P<0.05)。结论 CD137通过调控VSMC中NLRP3、IL-1β表达参与老年血管慢性炎症。Objective To investigate the mechanism of CD137 signaling involved in chronic inflammation of aging vessels.Methods Forty C57/B6 mice and twenty CD137^(-/-)mice were divided into control group, aging group and aging CD137^(-/-)group, with 20 mice each group.Luminex liquid chip multi-factor assay was used to detect the expression levels of inflammatory factors in the plasma of each group.IL-1β in the vessels was quantified and localized by q-PCR and immunofluorescence assay.Vascular smooth muscle cells were extracted and primarily cultured from the aorta of C57/B6 and CD137^(-/-)mice, respectively, and were divided into control group, aging group and aging CD137group.Cell senescence was induced by bleomycin treatment.The protein and RNA levels of NLRP3 and IL-β were detected by Western blotting and qPCR,respectively.The secretion level of IL-1β of cells was measured by ELISA.Results The plasma IL-1 β level was 10.86±0.89,61.40±3.88 and 18.20±1.25 pg/ml, in the control, aging and aging CD137^(-/-)group, respectively.The levels of plasma chemokine ligand 5,IL-3,IL-13,TNF-α and IL-1β were significantly higher in the aging group than the control group, while the levels in the aging CD137^(-/-)group were obviously lower than those in the aging group(P<0.01).The mRNA levels of vascular NLRP3 and IL-1 β were significantly higher in the aging group than the control group and the aging CD137^(-/-)group(3.45±0.62 vs 1.00±0.00 and 1.57±0.17,P<0.05;5.89±0.82 vs 1.00±0.00 and 2.64±0.34,P<0.05).In in vitro experiment, the protein expression of NLRP3 and IL-1 β in VSMC of the aging group was significantly higher than that of control group and that of aging CD137^(-/-)group(1.65±0.05 vs 1.00±0.00 and 0.85±0.05,P<0.05;1.45±0.05 vs 1.00±0.00 and 1.35±0.05,P<0.05).The level of IL-1 β in the supernatant of VSMC was significantly higher in the aging group than the control group and the aging CD137^(-/-)group(27.50±4.32 vs 14.20±3.52 and 16.50±2.72,P<0.05).Conclusion CD137 is involved in chronic inflammation

关 键 词：肌 平滑 血管 白细胞介素1Β 炎症 RNA 信使 巨噬细胞 NLR家族 热蛋白结构域包含蛋白3 

分 类 号：R543[医药卫生—心血管疾病]