中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2009年
8期
741-744
,共4页
庄欣琪%吕国义%杨阔%徐勇%邓迺封
莊訢琪%呂國義%楊闊%徐勇%鄧迺封
장흔기%려국의%양활%서용%산내봉
p38丝裂原活化蛋白激酶类%麻醉药,吸入%缺血后处理%心肌再灌注损伤
p38絲裂原活化蛋白激酶類%痳醉藥,吸入%缺血後處理%心肌再灌註損傷
p38사렬원활화단백격매류%마취약,흡입%결혈후처리%심기재관주손상
p38mitogen-activated protein kinases%Anesthetics,inhalation%Ischemie postcon-ditioning%Myocardial reperfusion injury
目的 评价p38丝裂原活化蛋白激酶(p38MAPK)信号转导通路在七氟烷后处理减轻乳鼠心肌细胞缺氧复氧损伤中的作用.方法 健康新生SD大鼠,日龄1～3 d,处死后取心室肌组织,培养心肌细胞,随机分为7组:对照组(C组)于CO2培养箱中持续培养3 h;缺氧复氧组(AR组)细胞缺氧2 h,复氧1 h;七氟烷后处理组(SP组)细胞缺氧2 h,复氧开始即刻更换为3%七氟烷饱和的DMEM培养液,孵育20 min,再更换为无血清DMEM培养液,继续复氧40 min;七氟烷后处理+SB203580组(SP+SB组)于七氟烷后处理同时加入SB203580(p38MAPK特异性抑制剂)至5 μmol/L,孵育20 min;七氟烷后处理+二甲亚砜组(SP+DMSO组)于七氟烷后处理同时加入0.1%DMSO,孵育20 min;SB203580组(SB组)于复氧开始时加入SB203580至5 μmol/L,孵育20 min;二甲亚砜组(DMSO组)于复氧开始即刻加入0.1%DMSO,孵育20 min.各组细胞分别接种于24孔培养板(1 ml/孔)、35 mm培养皿(5 mlnd/皿)和50 ml培养瓶(8 ml/瓶)中,每组12孔、6皿和6瓶.于复氧结束后,采用比色法测定细胞培养液乳酸脱氢酶(LDH)活性;采用台盼蓝排斥实验测定细胞存活率;采用流式细胞仪测定细胞凋亡率;采用Western blot法测定磷酸化p38MAPK(p-p38MAPK)表达水平.结果 与C组比较,其余各组LDH活性升高,细胞存活率降低,细胞凋亡率升高,AR组、SP组、SP+SB组、SP+DMSO组和DMSO组p-p38MAPK表达上调(P<0.05);与AR组比较,SP组、SP+SB组和SP+DMSO组LDH活性降低,细胞存活率升高,细胞凋亡率降低,p-p38MAPK表达上调(P<0.05);与SP组比较,SP+SB组、SB组和DMSO组LDH活性升高,细胞存活率降低,细胞凋亡率升高,p-p38MAPK表达下调(P<0.05).结论 七氟烷后处理可通过激活p38MAPK信号转导通路减轻乳鼠心肌细胞缺氧复氧损伤.
目的 評價p38絲裂原活化蛋白激酶(p38MAPK)信號轉導通路在七氟烷後處理減輕乳鼠心肌細胞缺氧複氧損傷中的作用.方法 健康新生SD大鼠,日齡1～3 d,處死後取心室肌組織,培養心肌細胞,隨機分為7組:對照組(C組)于CO2培養箱中持續培養3 h;缺氧複氧組(AR組)細胞缺氧2 h,複氧1 h;七氟烷後處理組(SP組)細胞缺氧2 h,複氧開始即刻更換為3%七氟烷飽和的DMEM培養液,孵育20 min,再更換為無血清DMEM培養液,繼續複氧40 min;七氟烷後處理+SB203580組(SP+SB組)于七氟烷後處理同時加入SB203580(p38MAPK特異性抑製劑)至5 μmol/L,孵育20 min;七氟烷後處理+二甲亞砜組(SP+DMSO組)于七氟烷後處理同時加入0.1%DMSO,孵育20 min;SB203580組(SB組)于複氧開始時加入SB203580至5 μmol/L,孵育20 min;二甲亞砜組(DMSO組)于複氧開始即刻加入0.1%DMSO,孵育20 min.各組細胞分彆接種于24孔培養闆(1 ml/孔)、35 mm培養皿(5 mlnd/皿)和50 ml培養瓶(8 ml/瓶)中,每組12孔、6皿和6瓶.于複氧結束後,採用比色法測定細胞培養液乳痠脫氫酶(LDH)活性;採用檯盼藍排斥實驗測定細胞存活率;採用流式細胞儀測定細胞凋亡率;採用Western blot法測定燐痠化p38MAPK(p-p38MAPK)錶達水平.結果 與C組比較,其餘各組LDH活性升高,細胞存活率降低,細胞凋亡率升高,AR組、SP組、SP+SB組、SP+DMSO組和DMSO組p-p38MAPK錶達上調(P<0.05);與AR組比較,SP組、SP+SB組和SP+DMSO組LDH活性降低,細胞存活率升高,細胞凋亡率降低,p-p38MAPK錶達上調(P<0.05);與SP組比較,SP+SB組、SB組和DMSO組LDH活性升高,細胞存活率降低,細胞凋亡率升高,p-p38MAPK錶達下調(P<0.05).結論 七氟烷後處理可通過激活p38MAPK信號轉導通路減輕乳鼠心肌細胞缺氧複氧損傷.
목적 평개p38사렬원활화단백격매(p38MAPK)신호전도통로재칠불완후처리감경유서심기세포결양복양손상중적작용.방법 건강신생SD대서,일령1～3 d,처사후취심실기조직,배양심기세포,수궤분위7조:대조조(C조)우CO2배양상중지속배양3 h;결양복양조(AR조)세포결양2 h,복양1 h;칠불완후처리조(SP조)세포결양2 h,복양개시즉각경환위3%칠불완포화적DMEM배양액,부육20 min,재경환위무혈청DMEM배양액,계속복양40 min;칠불완후처리+SB203580조(SP+SB조)우칠불완후처리동시가입SB203580(p38MAPK특이성억제제)지5 μmol/L,부육20 min;칠불완후처리+이갑아풍조(SP+DMSO조)우칠불완후처리동시가입0.1%DMSO,부육20 min;SB203580조(SB조)우복양개시시가입SB203580지5 μmol/L,부육20 min;이갑아풍조(DMSO조)우복양개시즉각가입0.1%DMSO,부육20 min.각조세포분별접충우24공배양판(1 ml/공)、35 mm배양명(5 mlnd/명)화50 ml배양병(8 ml/병)중,매조12공、6명화6병.우복양결속후,채용비색법측정세포배양액유산탈경매(LDH)활성;채용태반람배척실험측정세포존활솔;채용류식세포의측정세포조망솔;채용Western blot법측정린산화p38MAPK(p-p38MAPK)표체수평.결과 여C조비교,기여각조LDH활성승고,세포존활솔강저,세포조망솔승고,AR조、SP조、SP+SB조、SP+DMSO조화DMSO조p-p38MAPK표체상조(P<0.05);여AR조비교,SP조、SP+SB조화SP+DMSO조LDH활성강저,세포존활솔승고,세포조망솔강저,p-p38MAPK표체상조(P<0.05);여SP조비교,SP+SB조、SB조화DMSO조LDH활성승고,세포존활솔강저,세포조망솔승고,p-p38MAPK표체하조(P<0.05).결론 칠불완후처리가통과격활p38MAPK신호전도통로감경유서심기세포결양복양손상.
Objective To evalnate the role of p38 mitogen-activated protein kinase (p38MAPK) signal pathway in the protective effect of sevoflurane postconditioning (S-Postcon) on cultured neonatal rat cardiomyocytes against anoxia/reoxygenation (A/R) injury. Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into 7 groups: group Ⅰ normal control (C); group Ⅱ A/R; group Ⅲ S-Poatcan + A/R; group ⅣS-Postcon + SB203580 + A/R; group Ⅴ S-Postcon + DMSO + A/R; group Ⅵ SB203580 + A/R and group Ⅶ DMSO + A/R. GroupⅡ-Ⅶ were exposed to 2 h anoxia (95% N2-5% CO2) followed by 1 h reoxygenation. In group Ⅲ, Ⅳ and Ⅴ the cultured myocytes were postconditioned with 20 min 3% sevoflurane in 97% O2 alone (in group Ⅲ) or in conjunction with 5 μmol/L 5B203580 (a specific p38 MAPK inhibitor) (in group Ⅳ) or 0.1% DMSO (in group Ⅴ) followed by 40 min reoxygenation. The cardiomyocytes were reoxygenated in the presence of 5 μmol/L SB203580 in group Ⅵ or 0.1% DMSO in group Ⅶ. The LDH activity, cell survival rate and apoptotic rate were measured at the end of the experiment. The levels of phosphor-p38MAPK (p-p38MAPK) was detected by Western blotting. Results S-Postcon reduced LDH activity and apoptotic rate and increased cell survival rate and the level of p-p38MAPK as compared with group A/R (group Ⅱ). The myocardial protecfive effect of S-Posteon was eliminated by p38MAPK inhibltor-SB203580 and p-p38MAPK level was also decreased at the same time.Conclusion Sevoflurane postconditioning can attenuate anoxia/reoxygenation induced cardiomyocyte injury through activation of p38MAPK signal pathway.
