重组人白细胞介素-10对缺氧条件下人肺微血管内皮细胞凋亡的影响  

Effect of Recombinant Human Interleukin-10 on Hypoxia-Induced Apoptosis in Human Pulmonary Microvascular Endothelial Cells

作　　者：莫镜[1] 崔其亮[1] 姚君 张费通[1] 谭小华[1] 

机构地区：[1]广州医学院第三附属医院儿科,广州510150 [2]暨南大学第二临床医学院深圳市人民医院消化内科,广东深圳518020

出　　处：《实用儿科临床杂志》2012年第14期1107-1110,共4页Journal of Applied Clinical Pediatrics

基　　金：广东省科技计划项目(粤科规划字[2010]170号)

摘　　要：目的研究低氧条件下,重组人IL-10(rhIL-10)对人肺微血管内皮细胞(HPMVEC)凋亡和半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性的影响。方法常规培养HPMVEC细胞株,分为5组:正常对照组(常规培养);缺氧对照组(50 mL·L-1CO2,950mL·L-1N2);缺氧培养并加入rhIL-10,并按rhIL-10剂量(10×103ng.L-1、50×103ng.L-1、100×103ng.L-1)分为rhIL-10低剂量干预组、rhIL-10中剂量干预组、rhIL-10高剂量干预组。培养4 h、12 h、24 h、48 h离心收集细胞,化学比色法检测细胞核内Caspase-3活性,24 h核荧光染色并在荧光显微镜下观察荧光强度,并进行统计学分析。结果 4 h,凋亡蛋白Caspase-3相对活性各组间无明显差异(P>0.05);12 h,缺氧对照组,rhIL-10低、中剂量干预组与正常对照组比较显著升高(Pa<0.05),rhIL-10高剂量干预组与正常对照组比较无明显差异(P>0.05),rhIL-10各剂量干预组与缺氧对照组比较显著降低(Pa<0.05);24 h,缺氧对照组、rhIL-10各剂量干预组与正常对照组比较显著升高(Pa<0.05),rhIL-10各剂量干预组与缺氧对照组比较明显降低(Pa<0.05);48 h,缺氧对照组和rhIL-10低剂量干预组与正常对照组比较显著升高(Pa<0.05),低、中、高剂量rhIL-10干预组与缺氧对照组比较明显降低(Pa>0.05);缺氧凋亡峰值见于24 h。24 h,各组行凋亡细胞核Hoechst荧光染色,缺氧对照组和rhIL-10低、中、高剂量干预组灰度值与正常对照组比较均显著升高(Pa<0.05);rhIL-10低、中、高剂量rhIL-10干预组与缺氧对照组比较显著降低(Pa<0.05)。结论缺氧可促进HPMVEC凋亡,rhIL-10可一定程度地抑制HPMVEC凋亡,这些变化可能与急性缺氧性各器官疾病密切相关。Objective To investigate the ameliorative mechanism of recombinant human interleukin - 10 （ rhIL - 10） through inhibiting the apoptosis and Caspase - 3 activity in hypoxia - induced human pulmonary microvascular endothelial cells （ HPMVEC ）. Methods The cultured HPMVEC were randomly divided into normal control group,hypoxia control group,the low- rhIL- 10 interveation group,the middle- rhlL - 10 intervention group and the high - dose rhlL - 10 intervention group（ 10 ×10^3 ng·L-1 ,50×103 ng · L-l, 100×103 ng· L-1 ）. The normal group was cultured with the 37 ℃ and 50 mL· L-1COz and 210 mL· L-I O2 condition,and the hypoxis group was cultured with 50 mL· L -1 CO2 ,950 mL · L-I N2 condition. The culture time lasted for 4 h, 12 h ,24 h and 48 h ,respectively. The relative Caspase - 3 activity in all groups was detected with chromatometry at the end of 4 h, 12 h ,24 h and 48 h. The apoptosis fluorescence intensity was analyzed by using the fluorescent microscope at the end of 24 h with statistical method. Results There was no significant diffennce in the apoptosis and Caspase -3 activity among all the groups at the end of 4 h（ P 〉 0.05 ）. At the end of 12 h, the relative Caspase -3 activity was significantly increased in the hypoxia control group,the low dose and the middle dose rhIL- 10 intervention groups,compared to the normal control group （Pa 〈0.05）. The relative Caspase -3 activity in the high dose rhIL- 10 intervention groups was not obviously different from that in the nor- mal group （Pa 〉 0.05 ）, while it was relatively lower than that in the other groups （P 〈 0.05 ）. At the end of 24 h, the relative Caspase -3 activity in the hypoxia control group, the low dose, the middle dose and the high dose rhlL - 10 intervention groups were higher than that in the normal group （Pa 〈 0.05 ） ,while the relative Caspase -3 activity in the low ,the medial and the high dose rhIL- 10 intervention groups were lower than that in the hypoxia control group （P, 〈 0.05 ）. A

关 键 词：重组人白细胞介素-10 缺氧 凋亡 人肺微血管内皮细胞 半胱氨酸天冬氨酸蛋白酶-3 

分 类 号：R363[医药卫生—病理学]