加入结合胆红素与游离胆红素培养的人脐静脉内皮细胞增殖、迁移及血管生成能力比较  

Comparison of proliferation,migration,and angiogenesis between HUVECs added with conjugated bilirubin and unconjugated bilirubin

作　　者：周雪丽 李曼 季文斌 吕振宇 杨燕[1] 朱建[2] ZHOU Xueli;LI Man;JI Wenbin;LYU Zhenyu;YANG Yan;ZHU Jian(Department of Medical Oncology,The First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China;不详)

机构地区：[1]蚌埠医学院第一附属医院肿瘤内科,安徽蚌埠233004 [2]蚌埠医学院第一附属医院心内科

出　　处：《山东医药》2022年第23期36-39,共4页Shandong Medical Journal

基　　金：安徽省临床重点专科建设项目(皖卫科教秘2017-27号);蚌埠医学院自然科学重点项目(2020byzd056);蚌埠医学院“512人才培育计划”(by51201321);蚌埠医学院研究生科研创新计划(Byycx20046)。

摘　　要：目的 比较结合胆红素(CB)和游离胆红素(UCB)对人脐静脉内皮细胞(HUVECs)增殖、迁移和血管生成能力的影响,并探讨其潜在的机制。方法 取传代培养的HUVECs,随机分为三组,CB组加入20μmol/L CB、UCB组加入20μmol/L UCB、Control组加入最大药物浓度的DMSO(体积比≤0.1%)。采用MTS法检测三组细胞增殖能力(OD值),细胞划痕实验检测三组培养10、24 h的迁移能力(划痕修复率),成管实验检测三组培养4、8、24 h的血管生成能力(管样结构数量),Western blotting法检测三组细胞外信号调节激酶(ERK)、p-ERK1/2及蛋白激酶B(Akt)、pAkt308、p-Akt473蛋白表达。结果 UCB组、Control组、CB组OD值分别为0.48±0.05、0.81±0.02、1.56±0.07,三组OD值依次升高(P均<0.05)。与Control组比较,CB组培养10、24 h的划痕修复率及培养4、8、24 h的管样结构数量均升高,UCB组均降低(P均<0.05)。与Control组比较,CB组细胞ERK、Akt蛋白表达均无明显变化(P均>0.05),UCB组细胞ERK、Akt蛋白表达均升高(P均<0.05)。与Control组比较,CB组及UCB组细胞p-ERK1/2、p-Akt308、pAkt473蛋白表达均升高,且UCB组升高更明显(P均<0.05)。结论 CB可促进HUVECs的细胞增殖、迁移和血管生成能力,其机制可能与激活ERK1/2及Akt信号通路有关,而UCB对HUVECs的上述生物学行为则具有相反的作用。Objective To compare the effects of conjugated bilirubin(CB) and unconjugated bilirubin(UCB) on proliferation,migration and angiogenesis of human umbilical vein endothelial cells(HUVECs),and to explore their underlying mechanisms.Methods The subcultured HUVECs were randomly divided into three groups:the CB group(added with 20 μmol/L CB),the UCB group(added with 20 μmol/L UCB) and the Control group [added with the maximum drug concentration of DMSO(volume ratio≤0.1%)].The proliferation abilities of HUVECs in the three groups were detected by MTS assay(OD value);the migration abilities were detected by Scratch test at 10 and 24 h,and the angiogenesis abilities of them were determined by tube formation assay at 4,8 and 24 h.Furthermore,Western blotting was performed to detect the protein expression levels of ERK,p-ERK1/2,p-Akt^(308) and p-Akt^(473) of HUVECs.Results The OD values of the UCB group,the Control group and the CB group were 0.48±0.05,0.81±0.02 and 1.56±0.07,respectively,and the OD values of the three groups successively increased(all P<0.05).Compared with the Control group,the rates of scratch wound repair at 10 and 24 h and the number of tube-like structures at 4,8 and 24 h increased in the CB group,and decreased in the UCB group(all P<0.05).Compared with the control group,the CB group had no significant change in the protein expression of ERK or Akt(both P>0.05),but UCB group had increased protein expression of ERK and Akt(both P<0.05).Compared with the Control group,the protein expression of p-Akt^(308) and p-Akt^(473) increased in both CB and UCB groups,and the increase was more significant in the UCB group(both P<0.05).Conclusion CB can enhance the proliferation,migration and angiogenesis of HUVECs by activating ERK1/2 and Akt signaling pathways,whereas UCB have the opposite effects on above biological behaviors of HUVECs.

关 键 词：肿瘤学实验研究 结合胆红素 游离胆红素 人脐静脉内皮细胞 细胞增殖 细胞迁移 血管生成 ERK Akt 

分 类 号：R73-3[医药卫生—肿瘤]