摘要
目的研究CXCR3及IP-10对高糖诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)凋亡的影响,并对其机制进行初步探讨。方法 HUVEC在高糖和正常糖DMEM培养液中分别培养24 h、48 h、72 h,流式细胞术检测凋亡率,Western blot检测CXCR3表达。反义肽核酸(asPNA)阻断高糖组细胞表达CXCR3后,Western blot检测CXCR3表达,流式细胞术检测凋亡率。CXCR3配体IP-10及拮抗剂AMG487干预高糖组细胞48 h后,流式细胞术检测凋亡率。结果高糖组48 h、72 h时细胞凋亡率(22.56±1.83)%、(25.33±2.34)%较对照组(3.01±0.27)%、(3.80±0.32)%增加显著(均为P<0.01)。高糖组48 h、72 h CXCR3表达量0.57±0.04、0.87±0.03较对照组48 h 0.36±0.02、0.71±0.02明显上调(P<0.01、<0.05)。1μmol·L-1asPNA CXCR3组、2μmol·L-1asPNA CXCR3组CXCR3表达量0.32±0.02、0.14±0.02均较空白对照组(0.61±0.02)及阴性对照组(0.59±0.03)显著下降(均为P<0.01)。同时,1μmol·L-1asPNA CXCR3组、2μmol·L-1asPNA CXCR3组细胞凋亡率(15.33±1.05)%、(9.00±0.76)%也较空白对照组(22.98±1.92)%及阴性对照组(21.34±1.73)%明显下降(均为P<0.01)。IP-10组细胞凋亡率(38.05±2.88)%较高糖对照组(21.11±2.02)%、IP-10+AMG487组(20.80±2.17)%及AMG487组(19.54±1.93)%显著增加(均为P<0.01)。结论 CXCR3、IP-10促进高糖诱导的HUVEC凋亡,其促凋亡机制与IP-10/CXCR3轴有关。
Objective To investigate the effects of CXC chemokine receptor 3 （CXCR3） and interferon induced protein 10 （IP-10） on apoptosis of human umbilical vein endothelial cells（HUVEC） induced by high glucose, and preliminaxy probe the apoptosis mechanism. Methods HUVEC were cultured respectively in high glucose and normal glucose DMEM medium for 24 hours, 48 hours and 72 hours. Flow cytometry was used to detect the apoptotic rate. The expression of CXCR3 was tested by Western blot. The expression of CXCR3 was blocked by antiseuse peptide nucleic acid （asPNA） in high glucose group, then expression of CXCR3 was tested by Western blot, flow cytometry was used to detect the apoptotic rate. HUVEC cultured in high glucose group were intervened with CXCR3＇ s ligand （IP-10） and antagonist（ AMG487 ） for 48 hours, then flow cytometry was used to detect the apoptotie rate. Results The apoptotic rate of high glucose group at 48 hours （ 22.56 ± 1. 83 ） % and 72 hours （ 25.33 ± 2.34 ） % were significantly higher than （3.01 ±0.27）% and 72h group （3.80 ±0.32）% of normal glucose group （both P 〈0.01 ）. And CXCR3 expression of high glucose group at 48 hours （0.57 ± 0.04 ） and 72 hours （0.87 ± 0.03 ） were obviously higher than those of normal glucose group at48 hours （0.36±0.02） and72 hours （0.71 ±0.02） （P〈0.01, 〈 0.05）. CXCR3 expression of asPNA CXCR3 1 μmol · L^-1 （0.32 ± 0. 02） and 2 μmol ·L^-1 group （0.14 ±0.02） was drastically down-regulated （ all P 〈0.01 ）, compared with blank control （0.61 ±0. 02） and negative control group （0.59 ±0. 026）. At the same time,the apoptotic rate of asPNA CXCR3 1 μmol · L^-1 （ 15.33 ± 1.05 ）% and 2 μmol· L^-1 group （9.00 ± 0. 76）% was markedly lower than that of blank control （22. 98 ± 1.92）% and negative control group （21.34 ± 1.73 ）% （ all P 〈0.01 ）. The apoptotie rate of IP- 10 group （ 38.05 ± 2.88） % was significantly higher than that of high glucose group �
出处
《眼科新进展》
CAS
北大核心
2014年第7期628-632,共5页
Recent Advances in Ophthalmology
基金
上海市科委自然科学基金资助(编号:10ZR1418500)
上海市重点学科建设资助项目(编号:S30205)~~
