摘要
目的观察钠-钾-氯共转运体1(Na+-K+-2Cl-cotransporter 1,NKCC1)抑制剂布美他尼(Bumetanide)对脑微血管内皮细胞(BMEC)缺氧损伤后增殖的影响。方法将原代培养的大鼠BMEC分为3组:对照组、氧糖剥夺/复氧(OGD/Reo)组和OGD/Reo+Bumetanide(10μmol/L)组。在氧糖剥夺/复氧6、12、24和48h时,用MTT法检测细胞活力,Edu染色以及流式细胞术检测细胞增殖及细胞周期情况。结果氧糖剥夺/复氧6、12、24h,BMEC活力下降,Edu染色阳性率减少且S期细胞减少。而与OGD/Reo组相比,OGD/Reo+Bumetanide组BMEC活力增加[复氧6h:(0.497±0.039)vs.(0.612±0.044)],Edu染色阳性率增加[6h时:(5.742±0.195)%vs.(7.125±0.144)%],且S期细胞增多[6h时:(5.845±0.389)%vs.(7.124±0.542)%],差异均有统计学意义(均P<0.05)。结论布美他尼可促进缺氧损伤后BMEC的活力及细胞增殖,提示NKCC1通路在BMEC缺氧损伤后的细胞增殖中发挥了重要作用。
Objective To observe the effects of Bumetanide（NKCC1 inhibitor）on proliferation of brain microvascular endo‐thelial cells（BMEC）after oxygen glucose deprivation/reoxygenation（OGD/Reo）.Methods Primary BMEC were randomly di‐vided into three groups ：control group ,OGD/Reo group and OGD/Reo＋Bumetanide group. At different time points（6 ,12 ,24 , 48 h）of reoxygenation ,cell vitality was determined by MTT test.Edu dyeing and flow cytometry were used to detect the cell cy‐cle progress.Results After intervention with OGD/Reo for 6 ,12 and 24 h ,BMEC viability ,Edu positive rate and cells in S phase were decreased.Compared with the OGD/Reo group ,the cell vitality ,Edu positive rate and cells in S phase were signifi‐cantly increased [（0.497 ± 0.039） vs. （0.612 ± 0.044） ,（5.742 ± 0.195）% vs. （7.125 ± 0.144）% ,（5.845 ± 0.389）% vs. （7.124 ± 0.542）% ,all P＆lt; 0.05].Conclusion Bumetanide can increase BMEC vitality and promote cell proliferation after OGD ,suggesting NKCC1 plays an important role in proliferation of BMEC after OGD.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2014年第6期615-618,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81471200
No.81030021)
