白细胞介素-33抑制脂多糖诱导的心脏微血管内皮细胞通透性的研究初探  

Interleukin 33 inhibits lipopolysaccharide-induced high permeability of cardiac microvascular endothelial cells

作　　者：黄珊珊 杨舟鑫[2] 郭冬阳 贾兵兵[2] 严静[2] Huang Shanshan;Yang Zhouxin;Guo Dongyang;Jia Bingbing;Yan Jing(The Second Clinical Medical Collage,Zhejiang Chinese Medicine University,Hangzhou 310053,China;Department of Critical Care Medicine,Zhejiang Hospital,Hangzhou 310013,China)

机构地区：[1]浙江中医药大学第二临床学院,杭州310053 [2]浙江医院重症医学科,杭州310013

出　　处：《中华内科杂志》2022年第5期559-564,共6页Chinese Journal of Internal Medicine

基　　金：国家自然科学基金 (81772051)。

摘　　要：目的:探讨白细胞介素-33(IL-33)对脂多糖(LPS)诱导的大鼠心脏微血管内皮细胞(RCMECs)通透性的影响。方法:体外培养RCMECs,分为空白对照组、LPS组、IL-33组和LPS+IL-33组。用细胞计数试剂(CCK8)检测IL-33对RCMECs增殖的影响。异硫氰酸荧光素(FITC)-葡聚糖法检测4组单层RCMECs通透性。Western Blot检测4组RCMECs中血管内皮钙黏蛋白、Ras同源基因家族(Rho)成员A(RhoA)、磷酸化Rho相关含卷曲螺旋蛋白激酶(p-ROCK2)蛋白表达。高通量测序比较LPS组和LPS+IL-33组的差异基因表达,并对差异基因进行基因本体(GO)富集分析。结果:10~50 ng/ml的IL-33对RCMECs增殖无显著影响( P>0.05)。与空白对照组比,LPS组RCMECs通透性增加(相对灰度值1.404±0.029 比 1.000±0.200, P<0.05),血管内皮钙黏蛋白表达显著下调(相对灰度值0.429 5±0.012 9 比 0.594 9±0.014 2, P<0.05);而与LPS组比,LPS+IL-33组可降低RCMECs通透性(相对灰度值0.948±0.013, P<0.01),上调血管内皮钙黏蛋白表达(相对灰度值0.549 1±0.012 0, P<0.005)。与空白对照组比,LPS组RhoA(相对灰度值0.211 4±0.009 9 比 0.135 0±0.007 6, P<0.000 1)、p-ROCK(相对灰度值0.656 3±0.013 2 比 0.503 6±0.036 2, P<0.000 1,)蛋白表达上调;与LPS组比,LPS+IL-33组RhoA(相对灰度值 0.157 7±0.010 7, P=0.000 2)、p-ROCK(相对灰度值0.427 7±0.003 8, P<0.000 1)蛋白表达降低。高通量测序发现,LPS组、LPS+IL-33组下调的差异基因与细胞骨架及Rho信号通路相关。 结论:IL-33可能通过抑制Rho/Rho相关含卷曲螺旋蛋白激酶信号通路RhoA、p-ROCK蛋白表达,改善LPS诱导的心脏微血管内皮细胞的高通透性。Objective To investigate the effect of interleukin-33(IL-33)on lipopolysaccharide(LPS)-induced permeability of rat cardiac microvascular endothelial cells(RCMECs).Methods RCMECs were cultured in vitro to be divided into control group,LPS group,IL-33 group and LPS+IL-33 group.The effect of IL-33 on the proliferation of RCMECs was detected by cell counting reagent(CCK8).Fluorescein isothiocyanate(FITC)-dextran assay was used to evaluate the permeability of RCMECs.The expression of vascular endothelial calmodulin,ras homologous gene family(Rho)member A(RhoA)and phosphorylated Rho-associated coiled-coil-containing protein kinase(p-ROCK2)proteins were tested by western blot.High-throughput sequencing and gene ontology(GO)were performed for gene expression in LPS and LPS+IL-33 groups.Results No significant effect of IL-33 at 10-50 ng/ml on the proliferation of RCMECs was observed(P>0.05).Compared with the control group,the permeability of RCMECs(permeability coefficient ratio 1.404±0.029 vs.1.000±0.200,P<0.05)was significantly increased in LPS group and the expression of vascular endothelial calmodulin(relative gray value 0.4295±0.0129 vs.0.5949±0.0142,P<0.05)was down-regulated,while the permeability of monolayers(permeability coefficient ratio,0.948±0.013,P<0.01)was decreased in LPS+IL-33 group and the expression of vascular endothelial calmodulin(relative grayscale value 0.5491±0.0120,P<0.005)was up-regulated compared with the LPS group.High-throughput sequencing data revealed that the differential genes downregulated in the LPS and LPS+IL-33 groups were associated with cytoskeleton and Rho signaling pathway.Compared with the control group,RhoA(relative gray value 0.2114±0.0099 vs.0.1350±0.0076,P<0.0001)and p-ROCK(relative gray value 0.6563±0.0132 vs.0.5036±0.0362,P<0.0001)protein expression was upregulated in the LPS group.When compared with LPS group,RhoA(relative gray value 0.1577±0.0107,P=0.0002),p-ROCK(relative gray value 0.4277±0.0038,P<0.0001)protein expression was decreased in LPS+IL-33 group.Conc

关 键 词：白细胞介素33 脂多糖类 心脏微血管内皮细胞 通透性 

分 类 号：R459.7[医药卫生—急诊医学]