华中科技大学学报(医学版)
華中科技大學學報(醫學版)
화중과기대학학보(의학판)
ACTA UNIVERSITATIS MEDICINAE TONGJI
2014年
6期
615-618
,共4页
付佩彩%喻志源%李彩红%唐荣华%叶茂斌%王伟
付珮綵%喻誌源%李綵紅%唐榮華%葉茂斌%王偉
부패채%유지원%리채홍%당영화%협무빈%왕위
布美他尼%钠-钾-氯共转运体1%脑微血管内皮细胞%细胞增殖
佈美他尼%鈉-鉀-氯共轉運體1%腦微血管內皮細胞%細胞增殖
포미타니%납-갑-록공전운체1%뇌미혈관내피세포%세포증식
Bumetanide%Na+-K+-2Cl-cotransporter 1%brain microvascular endothelial cell%cell proliferation
目的:观察钠‐钾‐氯共转运体1(Na+‐K+‐2 Cl-cotransporter 1,NKCC1)抑制剂布美他尼(Bumetanide)对脑微血管内皮细胞(BM EC )缺氧损伤后增殖的影响。方法将原代培养的大鼠BM EC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48 h时,用M T T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、24 h ,BM EC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BM EC活力增加[复氧6 h:(0.497±0.039) v s.(0.612±0.044)],Edu染色阳性率增加[6 h时:(5.742±0.195)% v s.(7.125±0.144)%],且S期细胞增多[6 h时:(5.845±0.389)% v s.(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。
目的:觀察鈉‐鉀‐氯共轉運體1(Na+‐K+‐2 Cl-cotransporter 1,NKCC1)抑製劑佈美他尼(Bumetanide)對腦微血管內皮細胞(BM EC )缺氧損傷後增殖的影響。方法將原代培養的大鼠BM EC分為3組:對照組、氧糖剝奪/複氧(OGD/Reo )組和OGD/Reo+Bumetanide(10μmol/L )組。在氧糖剝奪/複氧6、12、24和48 h時,用M T T法檢測細胞活力,Edu染色以及流式細胞術檢測細胞增殖及細胞週期情況。結果氧糖剝奪/複氧6、12、24 h ,BM EC活力下降,Edu染色暘性率減少且S期細胞減少。而與OGD/Reo組相比,OGD/Reo+Bumetanide組BM EC活力增加[複氧6 h:(0.497±0.039) v s.(0.612±0.044)],Edu染色暘性率增加[6 h時:(5.742±0.195)% v s.(7.125±0.144)%],且S期細胞增多[6 h時:(5.845±0.389)% v s.(7.124±0.542)%],差異均有統計學意義(均 P<0.05)。結論佈美他尼可促進缺氧損傷後BMEC的活力及細胞增殖,提示NKCC1通路在BMEC缺氧損傷後的細胞增殖中髮揮瞭重要作用。
목적:관찰납‐갑‐록공전운체1(Na+‐K+‐2 Cl-cotransporter 1,NKCC1)억제제포미타니(Bumetanide)대뇌미혈관내피세포(BM EC )결양손상후증식적영향。방법장원대배양적대서BM EC분위3조:대조조、양당박탈/복양(OGD/Reo )조화OGD/Reo+Bumetanide(10μmol/L )조。재양당박탈/복양6、12、24화48 h시,용M T T법검측세포활력,Edu염색이급류식세포술검측세포증식급세포주기정황。결과양당박탈/복양6、12、24 h ,BM EC활력하강,Edu염색양성솔감소차S기세포감소。이여OGD/Reo조상비,OGD/Reo+Bumetanide조BM EC활력증가[복양6 h:(0.497±0.039) v s.(0.612±0.044)],Edu염색양성솔증가[6 h시:(5.742±0.195)% v s.(7.125±0.144)%],차S기세포증다[6 h시:(5.845±0.389)% v s.(7.124±0.542)%],차이균유통계학의의(균 P<0.05)。결론포미타니가촉진결양손상후BMEC적활력급세포증식,제시NKCC1통로재BMEC결양손상후적세포증식중발휘료중요작용。
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< 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.