中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2009年
4期
359-363
,共5页
杨孟昌%陈玉培%徐茜%曹德钧
楊孟昌%陳玉培%徐茜%曹德鈞
양맹창%진옥배%서천%조덕균
异氟醚%脂肪乳剂%静脉注射用%1-磷脂酰肌醇3-激酶%肌细胞%心脏%细胞低氧%氧
異氟醚%脂肪乳劑%靜脈註射用%1-燐脂酰肌醇3-激酶%肌細胞%心髒%細胞低氧%氧
이불미%지방유제%정맥주사용%1-린지선기순3-격매%기세포%심장%세포저양%양
Isoflurane%Fat emulsions,intravenous%1-Phosphatidylinositol 3-kinase%Myocytes,cardiac%Cell hypoxia%Oxygen
目的 评价磷脂酰肌醇-3激酶/蛋白质丝氨酸苏氨酸激酶(PI3K/Akt)信号通路在乳化异氟醚预先给药减轻乳鼠心肌细胞缺氧复氧损伤中的作用.方法 Wistar乳鼠60只,乳鼠心肌细胞于24孔培养板原代培养后,采用缺氧2 h复氧2 h建立缺氧复氧模型.乳鼠心肌细胞随机分为6组,每组8孔,正常对照组(C组):按正常条件继续培养4 h;A/R组:制备缺氧复氧模型;EI组:于细胞缺氧前即刻在培养液中加入乳化异氟醚,终浓度为1.68 mmol/L;EI+wortmannin(PI3K特异性抑制剂)组(EIW组):于细胞缺氧前即刻在培养液中加入乳化异氟醚和wortmannin,终浓度分别为1.68 mmol/L和100 nmol/L;wortmannin组(W组):于细胞缺氧前即刻在培养液中加入wortmannin,终浓度为100 nmol/L;脂肪乳组(F组):与细胞缺氧前即刻在培养液中加入30%脂肪乳,终浓度为0.05%.复氧2 h时,取细胞培养上清液测定乳酸脱氢酶(LDH)活性、肌钙蛋白I(cTnI)浓度,心肌细胞匀浆后测定超氧化物歧化酶(SOD)活性与丙二醛(MDA)含量,并测定心肌细胞磷酸化Akt(p-Akt)的表达水平.结果 与C组比较,其余5组上清液LDH活性、cTnI浓度和心肌细胞MDA含量升高,心肌细胞SOD活性降低(P<0.05);与A/R组比较,EI组和EIW组上清液LDH活性、cTnI浓度和心肌细胞MDA含量降低,心肌细胞SOD活性升高,F组除心肌细胞SOD活性升高外(P<0.05),其余指标差异均无统计学意义(P>0.05),W组上述指标差异无统计学意义(P>0.05);与EI组比较,EIW组、W组和F组上清液LDH活性、cTnI浓度和心肌细胞MDA含量升高,心肌细胞SOD活性降低(P<0.05).C组、EIW组、H/R组、F组和EI组心肌细胞p-Akt表达水平依次升高(P<0.05).结论 乳化异氟醚预先给药减轻乳鼠心肌细胞缺氧复氧损伤可能与进一步激活PI3K/Akt信号通路有关.
目的 評價燐脂酰肌醇-3激酶/蛋白質絲氨痠囌氨痠激酶(PI3K/Akt)信號通路在乳化異氟醚預先給藥減輕乳鼠心肌細胞缺氧複氧損傷中的作用.方法 Wistar乳鼠60隻,乳鼠心肌細胞于24孔培養闆原代培養後,採用缺氧2 h複氧2 h建立缺氧複氧模型.乳鼠心肌細胞隨機分為6組,每組8孔,正常對照組(C組):按正常條件繼續培養4 h;A/R組:製備缺氧複氧模型;EI組:于細胞缺氧前即刻在培養液中加入乳化異氟醚,終濃度為1.68 mmol/L;EI+wortmannin(PI3K特異性抑製劑)組(EIW組):于細胞缺氧前即刻在培養液中加入乳化異氟醚和wortmannin,終濃度分彆為1.68 mmol/L和100 nmol/L;wortmannin組(W組):于細胞缺氧前即刻在培養液中加入wortmannin,終濃度為100 nmol/L;脂肪乳組(F組):與細胞缺氧前即刻在培養液中加入30%脂肪乳,終濃度為0.05%.複氧2 h時,取細胞培養上清液測定乳痠脫氫酶(LDH)活性、肌鈣蛋白I(cTnI)濃度,心肌細胞勻漿後測定超氧化物歧化酶(SOD)活性與丙二醛(MDA)含量,併測定心肌細胞燐痠化Akt(p-Akt)的錶達水平.結果 與C組比較,其餘5組上清液LDH活性、cTnI濃度和心肌細胞MDA含量升高,心肌細胞SOD活性降低(P<0.05);與A/R組比較,EI組和EIW組上清液LDH活性、cTnI濃度和心肌細胞MDA含量降低,心肌細胞SOD活性升高,F組除心肌細胞SOD活性升高外(P<0.05),其餘指標差異均無統計學意義(P>0.05),W組上述指標差異無統計學意義(P>0.05);與EI組比較,EIW組、W組和F組上清液LDH活性、cTnI濃度和心肌細胞MDA含量升高,心肌細胞SOD活性降低(P<0.05).C組、EIW組、H/R組、F組和EI組心肌細胞p-Akt錶達水平依次升高(P<0.05).結論 乳化異氟醚預先給藥減輕乳鼠心肌細胞缺氧複氧損傷可能與進一步激活PI3K/Akt信號通路有關.
목적 평개린지선기순-3격매/단백질사안산소안산격매(PI3K/Akt)신호통로재유화이불미예선급약감경유서심기세포결양복양손상중적작용.방법 Wistar유서60지,유서심기세포우24공배양판원대배양후,채용결양2 h복양2 h건립결양복양모형.유서심기세포수궤분위6조,매조8공,정상대조조(C조):안정상조건계속배양4 h;A/R조:제비결양복양모형;EI조:우세포결양전즉각재배양액중가입유화이불미,종농도위1.68 mmol/L;EI+wortmannin(PI3K특이성억제제)조(EIW조):우세포결양전즉각재배양액중가입유화이불미화wortmannin,종농도분별위1.68 mmol/L화100 nmol/L;wortmannin조(W조):우세포결양전즉각재배양액중가입wortmannin,종농도위100 nmol/L;지방유조(F조):여세포결양전즉각재배양액중가입30%지방유,종농도위0.05%.복양2 h시,취세포배양상청액측정유산탈경매(LDH)활성、기개단백I(cTnI)농도,심기세포균장후측정초양화물기화매(SOD)활성여병이철(MDA)함량,병측정심기세포린산화Akt(p-Akt)적표체수평.결과 여C조비교,기여5조상청액LDH활성、cTnI농도화심기세포MDA함량승고,심기세포SOD활성강저(P<0.05);여A/R조비교,EI조화EIW조상청액LDH활성、cTnI농도화심기세포MDA함량강저,심기세포SOD활성승고,F조제심기세포SOD활성승고외(P<0.05),기여지표차이균무통계학의의(P>0.05),W조상술지표차이무통계학의의(P>0.05);여EI조비교,EIW조、W조화F조상청액LDH활성、cTnI농도화심기세포MDA함량승고,심기세포SOD활성강저(P<0.05).C조、EIW조、H/R조、F조화EI조심기세포p-Akt표체수평의차승고(P<0.05).결론 유화이불미예선급약감경유서심기세포결양복양손상가능여진일보격활PI3K/Akt신호통로유관.
Objective To evaluate the role of PI3K/Akt signal pathway in anoxia-reoxygenation (A/R) injury attenuated by emulsified isoflurane pretreatment in neonatal rat cardiomyocytes. Methods The cardiomyocytes of 60 neonatal Wistar rats (1-3 days old) were incubated in 24 well culture plates and randomly assigned into 6 groups (n=8 each): control group (group C);A/R group ;EI group;EI + wortmannin (specific inhibitor of PI3K) group (group EIW);wortmannin group (group W);fat emulsion group (group F). In group C, the cardiomyocytes were cultured for 4 h. In group A/R, the cardiomyocytes were exposed to 95% N2-5% CO2 saturated ischemic solution for 2 h followed by 2 h reoxygenation with 95 % O2-5% CO2 saturated reperfusion solution. In group EI, emulsified isoflurane was added to the culture medium immediately before cardiomyocyte hypoxia with the final concentration of 1.68 mmol/L. In group EIW, emulsified isoflurane and wortmannin were added to the culture medium in combination immediately before cardiomyocyte hypoxia with the final concentrations of 1.68 mmol/L and 100 nmol/L respectively. In group W and F, wortmannin and 30% fat emulsion were added to the culture medium immediately before cardiomyocyte hypoxia with the final concentration of 100 nmol/L and 0.05% respectively. The supernatant was taken from culture medium to measure LDH activity and cTnI concentration, and myocardial cell homogenate was prepared to measure SOD activity and MDA content and to detect phosphorylation-Akt (p-Akt) expression by Western blot at the end of 2 h reoxygenation. Results The supernatant LDH activity, cTnI concentration and cardiomyocyte MDA content were significantly higher, while SOD activity lower in the other 5 groups than in group C (P<0.05). The supernatant LDH activity, cTnI concentration and cardiomyocyte MDA content were significantly lower, while eardiomyocyte SOD activity higher in group EI and EIW, and cardiomyocyte SOD activity higher in group F than in group AIR (P<0.05), there was no significant difference in the above parameters between group W and A/R (P>0.05). The supernatant LDH activity, cTnI concentration and cardiomyocyte MDA content were significantly higher, while cardiomyocyte SOD lower in group EIW, W, and F than in group EI (P<0.05). The cardiomyocyte p-Akt expression was significantly increased in ascending order from group C, EIW, A/R, F to group EI (P<0.05). Conclusion Emulsified isoflurane pretreatment can attenuate A/R injury in neonatal rat cardiomyocytes via activation of PI3K/Akt signal pathway.