中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2013年
9期
1117-1120
,共4页
吴昊%何淑芳%朱海娟%金世云%胡军%张野
吳昊%何淑芳%硃海娟%金世雲%鬍軍%張野
오호%하숙방%주해연%금세운%호군%장야
受体,阿片样%1-磷脂酰肌醇3-激酶%蛋白质丝氨酸苏氨酸激酶%细胞外信号调节MAP激酶类%哌啶类%缺血预处理%细胞低氧
受體,阿片樣%1-燐脂酰肌醇3-激酶%蛋白質絲氨痠囌氨痠激酶%細胞外信號調節MAP激酶類%哌啶類%缺血預處理%細胞低氧
수체,아편양%1-린지선기순3-격매%단백질사안산소안산격매%세포외신호조절MAP격매류%고정류%결혈예처리%세포저양
Receptors,opioid%1-Phosphatidylinositol 3-kinase%Protein-serine-threonine kinases%Extracellular signal-regulated MAP kinases%Piperidines%Ischemic preconditioning%Cell hypoxia
目的 评价阿片受体、磷脂酰肌醇-3-激酶/丝氨酸-苏氨酸蛋白激酶(PI3K/Akt)和细胞外信号调节激酶(ERK)信号通路在瑞芬太尼预处理(RPC)减轻大鼠心肌细胞缺氧/复氧(H/R)损伤中的作用.方法 健康成年雄性SD大鼠,处死后取心室肌组织,原代培养心肌细胞,调整细胞浓度为2×104个/ml后接种于24孔细胞培养板(500 μl/孔)中,采用随机数字表法,将其中108个培养孔分为12组(n=9):正常对照组(C组)；H/R损伤组(H/R组)采用缺氧90 min/复氧120 min的方法制备心肌细胞H/R损伤模型；缺氧预处理组(HPC组)在H/R前,培养孔经缺氧10 min/复氧30 min；瑞芬太尼预处理组(RPC组)在H/R前,以终浓度1μmol/L瑞芬太尼孵育心肌细胞10 min,再常规培养30 min;δ受体拮抗剂纳曲吲哚+ RPC组(NTD+ RPC组)、κ受体拮抗剂nor-binahorphimine(nor-BNI)+ RPC组(BNI+RPC组)、PI3K/Akt信号通路阻断剂渥曼青霉素+RPC组(W+ RPC组)、ERK信号通路阻断剂PD98059+ RPC组(PD+ RPC组)在RPC前10 min分别维持培养孔终浓度5μmol/L纳曲吲哚和nor-BNI、0.1μmol/L渥曼青霉素和30 μmol/L PD98059,然后与瑞芬太尼共孵育10 min,再常规培养30 min,随后行H/R；试剂对照组分别为NTD组、BNI组、W组和PD组.于复氧结束时,测定心肌细胞活力、细胞凋亡率和培养液乳酸脱氢酶(LDH)活性.结果 与C组比较,H/R组心肌细胞活力降低,细胞凋亡率和培养液LDH活性升高(P＜0.05)；与H/R组比较,HPC组、RPC组和BNI+ RPC组心肌细胞活力升高,细胞凋亡率和培养液LDH活性降低(P＜0.05),NTD+ RPC组、W+ RPC组、PD+ RPC组、NTD组、BNI组、W组和PD组上述指标差异无统计学意义(P＞0.05)；与RPC组比较,NTD+ RPC组、BNI+ RPC组、W+ RPC组和PD+ RPC组心肌细胞活力降低,细胞凋亡率和培养液LDH活性升高(P＜0.05).结论 瑞芬太尼预处理可能通过激活δ受体,进而激活PI3K/Akt和ERK信号通路,从而减轻大鼠心肌细胞H/R损伤.
目的 評價阿片受體、燐脂酰肌醇-3-激酶/絲氨痠-囌氨痠蛋白激酶(PI3K/Akt)和細胞外信號調節激酶(ERK)信號通路在瑞芬太尼預處理(RPC)減輕大鼠心肌細胞缺氧/複氧(H/R)損傷中的作用.方法 健康成年雄性SD大鼠,處死後取心室肌組織,原代培養心肌細胞,調整細胞濃度為2×104箇/ml後接種于24孔細胞培養闆(500 μl/孔)中,採用隨機數字錶法,將其中108箇培養孔分為12組(n=9):正常對照組(C組)；H/R損傷組(H/R組)採用缺氧90 min/複氧120 min的方法製備心肌細胞H/R損傷模型；缺氧預處理組(HPC組)在H/R前,培養孔經缺氧10 min/複氧30 min；瑞芬太尼預處理組(RPC組)在H/R前,以終濃度1μmol/L瑞芬太尼孵育心肌細胞10 min,再常規培養30 min;δ受體拮抗劑納麯吲哚+ RPC組(NTD+ RPC組)、κ受體拮抗劑nor-binahorphimine(nor-BNI)+ RPC組(BNI+RPC組)、PI3K/Akt信號通路阻斷劑渥曼青黴素+RPC組(W+ RPC組)、ERK信號通路阻斷劑PD98059+ RPC組(PD+ RPC組)在RPC前10 min分彆維持培養孔終濃度5μmol/L納麯吲哚和nor-BNI、0.1μmol/L渥曼青黴素和30 μmol/L PD98059,然後與瑞芬太尼共孵育10 min,再常規培養30 min,隨後行H/R；試劑對照組分彆為NTD組、BNI組、W組和PD組.于複氧結束時,測定心肌細胞活力、細胞凋亡率和培養液乳痠脫氫酶(LDH)活性.結果 與C組比較,H/R組心肌細胞活力降低,細胞凋亡率和培養液LDH活性升高(P＜0.05)；與H/R組比較,HPC組、RPC組和BNI+ RPC組心肌細胞活力升高,細胞凋亡率和培養液LDH活性降低(P＜0.05),NTD+ RPC組、W+ RPC組、PD+ RPC組、NTD組、BNI組、W組和PD組上述指標差異無統計學意義(P＞0.05)；與RPC組比較,NTD+ RPC組、BNI+ RPC組、W+ RPC組和PD+ RPC組心肌細胞活力降低,細胞凋亡率和培養液LDH活性升高(P＜0.05).結論 瑞芬太尼預處理可能通過激活δ受體,進而激活PI3K/Akt和ERK信號通路,從而減輕大鼠心肌細胞H/R損傷.
목적 평개아편수체、린지선기순-3-격매/사안산-소안산단백격매(PI3K/Akt)화세포외신호조절격매(ERK)신호통로재서분태니예처리(RPC)감경대서심기세포결양/복양(H/R)손상중적작용.방법 건강성년웅성SD대서,처사후취심실기조직,원대배양심기세포,조정세포농도위2×104개/ml후접충우24공세포배양판(500 μl/공)중,채용수궤수자표법,장기중108개배양공분위12조(n=9):정상대조조(C조)；H/R손상조(H/R조)채용결양90 min/복양120 min적방법제비심기세포H/R손상모형；결양예처리조(HPC조)재H/R전,배양공경결양10 min/복양30 min；서분태니예처리조(RPC조)재H/R전,이종농도1μmol/L서분태니부육심기세포10 min,재상규배양30 min;δ수체길항제납곡신타+ RPC조(NTD+ RPC조)、κ수체길항제nor-binahorphimine(nor-BNI)+ RPC조(BNI+RPC조)、PI3K/Akt신호통로조단제악만청매소+RPC조(W+ RPC조)、ERK신호통로조단제PD98059+ RPC조(PD+ RPC조)재RPC전10 min분별유지배양공종농도5μmol/L납곡신타화nor-BNI、0.1μmol/L악만청매소화30 μmol/L PD98059,연후여서분태니공부육10 min,재상규배양30 min,수후행H/R；시제대조조분별위NTD조、BNI조、W조화PD조.우복양결속시,측정심기세포활력、세포조망솔화배양액유산탈경매(LDH)활성.결과 여C조비교,H/R조심기세포활력강저,세포조망솔화배양액LDH활성승고(P＜0.05)；여H/R조비교,HPC조、RPC조화BNI+ RPC조심기세포활력승고,세포조망솔화배양액LDH활성강저(P＜0.05),NTD+ RPC조、W+ RPC조、PD+ RPC조、NTD조、BNI조、W조화PD조상술지표차이무통계학의의(P＞0.05)；여RPC조비교,NTD+ RPC조、BNI+ RPC조、W+ RPC조화PD+ RPC조심기세포활력강저,세포조망솔화배양액LDH활성승고(P＜0.05).결론 서분태니예처리가능통과격활δ수체,진이격활PI3K/Akt화ERK신호통로,종이감경대서심기세포H/R손상.
Objective To evaluate the role of opioid receptors and phosphatidylinositol 3-kinase/proteinserine-threonine kinases (PI3K/Akt) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of hypoxia/reoxygenation (H/R)-induced injury to cardiomyocytes by remifentanil preconditioning in rats.Methods Primary cardiomyocytes were obtained from adult male Sprague-Dawley rats and cultured in DMEM culture medium.The cells were seeded in 48-well plates (density 2 × 104 cells/ml,500 μl/well) and randomly divided into 12 groups (n =9 each):control group (group C),group H/R,hypoxia preconditioning group (group HPC),remifentanil preconditioning (RPC) group,naltrindole (δ receptor antagonist) + RPC group,nor-binaltorphimine (κ receptor antagonist) + RPC group (BNI + RPC group),wortmannin (PI3K inhibitor) + RPC group (W+ RPC group),PD98059 (ERK inhibitor) + RPC group (PD + RPC group),NTD group,BNI group,W group and PD group.In group H/R,the cardiomyocytes were exposed to 90 min of hypoxia,followed by 120 min of reoxygenation.In group HPC,the cardiomyocytes were exposed to 10 min of hypoxia,followed by 30 min of reoxygenation before H/R.In group RPC,the cardiomyocytes were preconditioned with remifentanil with the final concentration of 1 μmol/L for 10 min,followed by 30 min routine culture before H/R.In NTD + RPC,BNI + RPC,W + RPC and PD + RPC groups,naltrindole 5μmol/L (final concentration),nor-binaltorphimine 5 μmol/L (final concentration),wortmannin 0.1 μmol/L (final concentration) and PD98059 30μmol/L (final concentration)were added,respectively,and then the cells were coincubated with remifentanil for 10 min,followed by 30 min routine culture before H/R.The viability of cardiomyocytes,cell apoptosis and activity of lactate dehydrogenase (LDH) in the culture medium were detected.The apoptosis rate (AR) was calculated.Results Compared with group C,the viability of cardiomyocytes,AR and activity of LDH in the culture medium were significantly increased in group H/R (P ＜ 0.05).Compared with group H/R,the viability of cardiomyocytes,AR and activity of LDH in the culture medium were significantly decreased in HPC,RPC and BNI + RPC groups (P ＜ 0.05),and no significant changes were found in the parameters mentioned above in NTD + RPC,W + RPC,PD + RPC,NTD,BNI,W,and PD groups (P ＞ 0.05).The viability of cardiomyocytes was significantly lower,and the AR and activity of LDH in the culture medium were higher in NTD + RPC,BNI + RPC,W + RPC,and PD + RPC groups than in RPC group (P ＜ 0.05).Conclusion Remifentanil preconditioning activates PI3K/Akt and ERK signaling pathways possibly through activating δ opioid receptors thus attenuating H/R-induced injury to cardiomyocytes in rats.
