中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2011年
1期
102-104
,共3页
陈瑾%曾苏%康焕菊%沈施仁
陳瑾%曾囌%康煥菊%瀋施仁
진근%증소%강환국%침시인
二氮嗪%心脏%内皮,血管%氧%细胞低氧%缺氧诱导因子-1α亚基%肿瘤抑制蛋白质p53
二氮嗪%心髒%內皮,血管%氧%細胞低氧%缺氧誘導因子-1α亞基%腫瘤抑製蛋白質p53
이담진%심장%내피,혈관%양%세포저양%결양유도인자-1α아기%종류억제단백질p53
Dazoxide%Heart%Endothelium,vascular%Oxygen%Cell hpoxia%Hypoxia-inducible factor 1,alpha subunit%Tumor suppressor protein p53
目的 探讨二氮嚓预先给药对大鼠心肌微血管内皮细胞缺氧复氧时缺氧诱导因子-1α(HIF-1α)mRNA和p53 mRNA表达的影响.方法 培养SD大鼠心肌微血管内皮细胞,以1×106/ml的密度接种于96孔板(100μl/孔)或培养皿(2 ml/皿),采用随机数字表法,将其随机分为4组(n=24):正常对照组(C组)不作任何处理,缺氧复氧组(H/R组)、二氮嗪预先给药组(DZ组)、二氮嗪预先给药+线粒体ATP敏感性钾通道阻断剂5-羟葵酸组(DZ+5-HD组)均进行缺氧2 h复氧2 h.DZ组和DZ+5-HD组在缺氧前2 h分别加入100μmol/L二氮嗪、100μmol/L二氮嗪+100μmol/L 5-羟葵酸.于复氧2h时测定细胞活力、细胞凋亡率、HIF-1α mRNA和p53 mRNA表达水平.结果 与C组比较,H/R组细胞活力降低,细胞凋亡率升高,HIF-1αmRNA和p53 mRNA表达上调(P＜0.01);与H/R组比较,DZ组细胞活力升高,细胞凋亡率降低,HIF-1αmRNA表达上调,p53 mRNA表达下调(P＜0.05或0.01);5-羟葵酸可抑制二氮嗪预先给药导致的上述改变(P＜0.05).结论 二氮嗪预先给药可上调HIF-1和下调p53表达,从而减轻大鼠心肌微血管内皮细胞缺氧复氧损伤,其机制与激活线粒体ATP敏感性钾通道有关.
目的 探討二氮嚓預先給藥對大鼠心肌微血管內皮細胞缺氧複氧時缺氧誘導因子-1α(HIF-1α)mRNA和p53 mRNA錶達的影響.方法 培養SD大鼠心肌微血管內皮細胞,以1×106/ml的密度接種于96孔闆(100μl/孔)或培養皿(2 ml/皿),採用隨機數字錶法,將其隨機分為4組(n=24):正常對照組(C組)不作任何處理,缺氧複氧組(H/R組)、二氮嗪預先給藥組(DZ組)、二氮嗪預先給藥+線粒體ATP敏感性鉀通道阻斷劑5-羥葵痠組(DZ+5-HD組)均進行缺氧2 h複氧2 h.DZ組和DZ+5-HD組在缺氧前2 h分彆加入100μmol/L二氮嗪、100μmol/L二氮嗪+100μmol/L 5-羥葵痠.于複氧2h時測定細胞活力、細胞凋亡率、HIF-1α mRNA和p53 mRNA錶達水平.結果 與C組比較,H/R組細胞活力降低,細胞凋亡率升高,HIF-1αmRNA和p53 mRNA錶達上調(P＜0.01);與H/R組比較,DZ組細胞活力升高,細胞凋亡率降低,HIF-1αmRNA錶達上調,p53 mRNA錶達下調(P＜0.05或0.01);5-羥葵痠可抑製二氮嗪預先給藥導緻的上述改變(P＜0.05).結論 二氮嗪預先給藥可上調HIF-1和下調p53錶達,從而減輕大鼠心肌微血管內皮細胞缺氧複氧損傷,其機製與激活線粒體ATP敏感性鉀通道有關.
목적 탐토이담찰예선급약대대서심기미혈관내피세포결양복양시결양유도인자-1α(HIF-1α)mRNA화p53 mRNA표체적영향.방법 배양SD대서심기미혈관내피세포,이1×106/ml적밀도접충우96공판(100μl/공)혹배양명(2 ml/명),채용수궤수자표법,장기수궤분위4조(n=24):정상대조조(C조)불작임하처리,결양복양조(H/R조)、이담진예선급약조(DZ조)、이담진예선급약+선립체ATP민감성갑통도조단제5-간규산조(DZ+5-HD조)균진행결양2 h복양2 h.DZ조화DZ+5-HD조재결양전2 h분별가입100μmol/L이담진、100μmol/L이담진+100μmol/L 5-간규산.우복양2h시측정세포활력、세포조망솔、HIF-1α mRNA화p53 mRNA표체수평.결과 여C조비교,H/R조세포활력강저,세포조망솔승고,HIF-1αmRNA화p53 mRNA표체상조(P＜0.01);여H/R조비교,DZ조세포활력승고,세포조망솔강저,HIF-1αmRNA표체상조,p53 mRNA표체하조(P＜0.05혹0.01);5-간규산가억제이담진예선급약도치적상술개변(P＜0.05).결론 이담진예선급약가상조HIF-1화하조p53표체,종이감경대서심기미혈관내피세포결양복양손상,기궤제여격활선립체ATP민감성갑통도유관.
Objective To investigate the effects of diazoxide pretreatment on the expression of hypoxia-inducible factor-1α (HIF-1α) mRNA and p53 mRNA in rat myocardial microvascular endothelial cells exposed to hypoxia-reoxygenation (H/R).Methods The SD rat myocardial microvascular endothelial cells were cultured. The cells were seeded in 96-well plates ( 100 μl/hole) or in 6 cm diameter dishes (2 ml/dish) with the density of 1 ×106/ml and randomly divided into 4 groups ( n = 24 each): normal control group (group C), H/R group, H/R +diazoxide pretreatment group (group DZ) and H/R + diazoxide pretreatment + mitochondrial ATP-sensitive potassium channel blocker 5-hydroxydecanoate (5-HD) group (group DZ + 5-HD). The cells were exposed to 2 h hypoxia followed by 2 h reoxygenation. Diazoxide 100 μmol/L and diazoxide 100 μmol/L + 5-HD 100 μmol/L were added to the culture medium 2 h before hypoxia in DZ and DZ + 5-HD groups respectively. The cell vitality, apoptotic rate and expression of HIF-1α mRNA and p53 mRNA were detected at the end of reoxygenation. Results Compared with group C, the cell vitality was significantly decreased, apoptotic rate increased and the expression of HIF- 1α mRNA and p53 mRNA up-regulated in H/R group ( P ＜ 0.01). Compared with group H/R, the cell vitality was significantly increased, apoptotic rate decreased, the expression of HIF-1α mRNA up-regulated and the expression of p53 mRNA down-regulated in group DZ ( P ＜ 0.05 or 0.01 ). 5-HD could inhibit diazoxide pretreatment-induced changes mentioned above (P ＜ 0.05 ). Conclusion Diazoxide pretreatment can reduce H/R injury in rat myocardial microvascular endothelial cells through up-regulating the expression of HIF-1α and down-regulating the expression of p53, and the mechanism is related to activation of mitochondrial ATP-sensitive potassium channels.
