中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2011年
8期
1001-1004
,共4页
孙志鹏%罗婷%黄德樱%乐江
孫誌鵬%囉婷%黃德櫻%樂江
손지붕%라정%황덕앵%악강
血红素加氧酶-1%细胞低氧%氧%肌细胞,心脏%七氟醚
血紅素加氧酶-1%細胞低氧%氧%肌細胞,心髒%七氟醚
혈홍소가양매-1%세포저양%양%기세포,심장%칠불미
Hene oxygenase-1%Cell hypoxia%Oxygen%Myocytes,cardiac%Sevoflurane
目的 探讨血红素加氧酶-1(HO-1)在七氟醚预处理减轻乳鼠心肌细胞缺氧复氧损伤中的作用.方法 新生健康清洁级SD大鼠15只,日龄1～3d,处死后取心室肌组织,原代培养心肌细胞,以1×106个/ml接种于6孔培养板或以2× 105个/ml接种于24孔培养板,采用随机数字表法,将其随机分为4组(n＝25):对照组(C组)常规培养；缺氧复氧组(H/R组)采用缺氧2h,复氧1h的方法制备心肌细胞缺氧复氧损伤模型；七氟醚预处理组(S+ H/R组)细胞经2.5％七氟醚预处理20min后行药物洗脱10 min,再行缺氧复氧处理；锌原卟啉+七氟醚预处理组(ZnPP+ S+ H/R组)细胞经HO-1抑制剂锌原卟啉3 μmol/L孵育1h后,行七氟醚预处理及缺氧复氧处理.于复氧结束后测定心肌细胞HO-1表达、细胞凋亡率、细胞内游离Ca2+浓度([Ca2+]i)、线粒体膜通透性转运孔(PTP)开放程度、细胞色素C(Cyto C)表达及培养液LDH和CK活性.结果 与C组比较,H/R组心肌细胞HO-1和胞浆Cyto C表达上调,线粒体Cyto C表达下调,培养液LDH、CK活性、细胞凋亡率、[Ca2+]i和PTP开放度升高(P<0.01).与H/R组比较,S+H/R组心肌细胞HO-1和线粒体Cyto C表达上调,胞浆Cyto C 表达下调,培养液LDH、CK活性、细胞凋亡率、[Ca2+]i和PIP开放度降低(P<0.01).与S+H/R组比较,ZnPP+ S+ H/R组心肌细胞HO-1和线粒体Cyto C表达下调,胞浆CytoC表达上调,培养液LDH、CK活性、细胞凋亡率、[Ca2+]i和PTP开放度升高(P<0.01).结论 HO-1表达上调参与了七氟醚预处理减轻乳鼠心肌细胞缺氧复氧损伤.
目的 探討血紅素加氧酶-1(HO-1)在七氟醚預處理減輕乳鼠心肌細胞缺氧複氧損傷中的作用.方法 新生健康清潔級SD大鼠15隻,日齡1～3d,處死後取心室肌組織,原代培養心肌細胞,以1×106箇/ml接種于6孔培養闆或以2× 105箇/ml接種于24孔培養闆,採用隨機數字錶法,將其隨機分為4組(n＝25):對照組(C組)常規培養；缺氧複氧組(H/R組)採用缺氧2h,複氧1h的方法製備心肌細胞缺氧複氧損傷模型；七氟醚預處理組(S+ H/R組)細胞經2.5％七氟醚預處理20min後行藥物洗脫10 min,再行缺氧複氧處理；鋅原卟啉+七氟醚預處理組(ZnPP+ S+ H/R組)細胞經HO-1抑製劑鋅原卟啉3 μmol/L孵育1h後,行七氟醚預處理及缺氧複氧處理.于複氧結束後測定心肌細胞HO-1錶達、細胞凋亡率、細胞內遊離Ca2+濃度([Ca2+]i)、線粒體膜通透性轉運孔(PTP)開放程度、細胞色素C(Cyto C)錶達及培養液LDH和CK活性.結果 與C組比較,H/R組心肌細胞HO-1和胞漿Cyto C錶達上調,線粒體Cyto C錶達下調,培養液LDH、CK活性、細胞凋亡率、[Ca2+]i和PTP開放度升高(P<0.01).與H/R組比較,S+H/R組心肌細胞HO-1和線粒體Cyto C錶達上調,胞漿Cyto C 錶達下調,培養液LDH、CK活性、細胞凋亡率、[Ca2+]i和PIP開放度降低(P<0.01).與S+H/R組比較,ZnPP+ S+ H/R組心肌細胞HO-1和線粒體Cyto C錶達下調,胞漿CytoC錶達上調,培養液LDH、CK活性、細胞凋亡率、[Ca2+]i和PTP開放度升高(P<0.01).結論 HO-1錶達上調參與瞭七氟醚預處理減輕乳鼠心肌細胞缺氧複氧損傷.
목적 탐토혈홍소가양매-1(HO-1)재칠불미예처리감경유서심기세포결양복양손상중적작용.방법 신생건강청길급SD대서15지,일령1～3d,처사후취심실기조직,원대배양심기세포,이1×106개/ml접충우6공배양판혹이2× 105개/ml접충우24공배양판,채용수궤수자표법,장기수궤분위4조(n＝25):대조조(C조)상규배양；결양복양조(H/R조)채용결양2h,복양1h적방법제비심기세포결양복양손상모형；칠불미예처리조(S+ H/R조)세포경2.5％칠불미예처리20min후행약물세탈10 min,재행결양복양처리；자원계람+칠불미예처리조(ZnPP+ S+ H/R조)세포경HO-1억제제자원계람3 μmol/L부육1h후,행칠불미예처리급결양복양처리.우복양결속후측정심기세포HO-1표체、세포조망솔、세포내유리Ca2+농도([Ca2+]i)、선립체막통투성전운공(PTP)개방정도、세포색소C(Cyto C)표체급배양액LDH화CK활성.결과 여C조비교,H/R조심기세포HO-1화포장Cyto C표체상조,선립체Cyto C표체하조,배양액LDH、CK활성、세포조망솔、[Ca2+]i화PTP개방도승고(P<0.01).여H/R조비교,S+H/R조심기세포HO-1화선립체Cyto C표체상조,포장Cyto C 표체하조,배양액LDH、CK활성、세포조망솔、[Ca2+]i화PIP개방도강저(P<0.01).여S+H/R조비교,ZnPP+ S+ H/R조심기세포HO-1화선립체Cyto C표체하조,포장CytoC표체상조,배양액LDH、CK활성、세포조망솔、[Ca2+]i화PTP개방도승고(P<0.01).결론 HO-1표체상조삼여료칠불미예처리감경유서심기세포결양복양손상.
Objective To investigate the role of heme oxygenase-1 (HO-1 ) in process of sevoflurane preconditioning attenuating hypoxia-reoxygenation (H/R) injury in cultured neonatal rat cardiomyocytes.Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into four groups ( n ＝ 25 each):control group (group C),group H/R,sevoflurane preconditioning group(group S + H/R),and HO-1 inhibitor zinc protoporphyria (ZnPP) and sevoflurane preconditioning group(group ZnPP + S + H/R).In group H/R,the cardiomyocytes were exposed to 2 hours of hypoxia,followed by 1 hour of reoxygenation.Group S+ H/R received 2.5％ sevoflurane preconditioning for 20 minutes followed by 10 minutes of wash-out before H/R.ZnPP was added to the culture medium with final concentrations of 3 μMol/L 1 hour before sevoflurane preconditioning and H/R in group ZnPP +S + H/R.HO- 1 expression,apoptosis rate,concentration of free calcium ( [ Ca2 + ] i),mitochondrial membrane permeability transition pore (PTP),cytoohrome C ( Cyto C) expression and activities of lactate dehydrogenase (LDH) and creatine kinase (CK) in culture supernatant were detected after reoxygenation.Results Compared with group C,the expression of HO-1 and cytoplasmic Cyto C of cardiomyocytes were up-regulated,mitochondrial Cyto C was down-regulated,while the [Ca2+ ]i,opening degree of PIP,apoptosis rate and activities of LDH and CK in culture supernatant were increased in group H/R.Compared with group H/R,the expression of HO-1 and mitochondrial Cyto C of cardiomyocytes were up-regulated,cytoplasmic Cyto C was down-regulated,while the [Ca2+ ] i,opening degree of PTP,apoptosis rate and activities of LDH and CK in culture supernatant were decreased in group S + H/R.Compared with group S + H/R,the expression of HO-1 and mitochondrial Cyto C of cardiomyocytes were down-regulated,cytoplasmic Cyto C was up-regulated,while the [Ca2+ ]i,opening degree of PTP,apoptosis rate and activities of LDH and CK in culture supematant were increased in group ZnPP + S + H/R.Conclusion The up-regulation of HO-1 is involved in the process of sevoflurane preconditioning attenuating hypoxia-reoxygenation injury in cultured neonatal rat cardiomyocytes.
