中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
Chinese Journal of Anesthesiology
2015年
9期
1050-1053
,共4页
杨立斌%摆志霞%吕丹妮%刘海波%陈学新
楊立斌%襬誌霞%呂丹妮%劉海波%陳學新
양립빈%파지하%려단니%류해파%진학신
线粒体膜转运蛋白质类%脂肪乳剂,静脉注射用%布比卡因%药物毒性%心肌
線粒體膜轉運蛋白質類%脂肪乳劑,靜脈註射用%佈比卡因%藥物毒性%心肌
선립체막전운단백질류%지방유제,정맥주사용%포비잡인%약물독성%심기
Mitochondrial membrane transport proteins%Fat emulsions,intravenous%Bupivacaine%Drug toxicity%Myocardium
目的 评价线粒体通透性转运孔在脂肪乳逆转布比卡因大鼠心肌毒性中的作用.方法 培养H9c2心肌细胞,以105个/ml的密度接种于6孔板,采用随机数字表法分为4组,每组6孔,每孔2 ml:对照组(C组)加入PBS 100μl;布比卡因组(B组)加入布比卡因,终浓度为1 mmol/L;脂肪乳+布比卡因组(LB组)同时加入脂肪乳和布比卡因,终浓度分别为1％和1 mmol/L;脂肪乳+布比卡因+苍术苷组(LBA组)同时加入脂肪乳、布比卡因和线粒体通透性转换孔开放剂苍术苷,终浓度分别为1％、1 mmol/L和30 μmol/L,均孵育24 h.孵育结束后,采用Western blot法测定Bcl-2、Bax、磷酸化Bad (p-Bad)、caspase-3、活化的caspase-3、caspase-9、活化的caspase-9和细胞色素c(Cyt c)的表达水平,采用RT-PCR法检测Bcl-2 mRNA、Bax mRNA、Bad mRNA、caspase-9 mRNA和Cyt c mRNA的表达水平.结果 与C组比较,B组Bax/Bcl-2比值、活化的caspase-3/caspase-3比值、活化的caspase-9/caspase-9比值和Bax mRNA/Bcl-2 mRNA比值升高,p-Bad表达下调,Cyt c、Bad mRNA、caspase-9mRNA和Cyt c mRNA的表达上调(P＜0.05),LB组上述各指标差异无统计学意义(P＞0.05);与B组比较,LB组和LBA组Bax/Bcl-2比值、活化的caspase-3/caspase-3比值和活化、caspase-9/caspase-9比值和Bax mRNA/Bcl-2 mRNA比值降低,p-Bad表达上调,Cyt c、Bad mRNA、caspase-9 mRNA和Cyt cmRNA的表达下调(P＜0.05);与LB组比较,LBA组Bax/Bcl-2比值、活化的caspase-3/caspase-3比值、活化的caspase-9/caspase-9比值和Bax mRNA/Bcl-2 mRNA比值升高,p-Bad表达下调,Cyt c、BadmRNA、caspase-9 mRNA和Cyt c mRNA的表达上调(P＜0.05).结论 脂肪乳逆转布比卡因大鼠心肌毒性作用的机制与抑制线粒体通透性转换孔的开放有关.
目的 評價線粒體通透性轉運孔在脂肪乳逆轉佈比卡因大鼠心肌毒性中的作用.方法 培養H9c2心肌細胞,以105箇/ml的密度接種于6孔闆,採用隨機數字錶法分為4組,每組6孔,每孔2 ml:對照組(C組)加入PBS 100μl;佈比卡因組(B組)加入佈比卡因,終濃度為1 mmol/L;脂肪乳+佈比卡因組(LB組)同時加入脂肪乳和佈比卡因,終濃度分彆為1％和1 mmol/L;脂肪乳+佈比卡因+蒼術苷組(LBA組)同時加入脂肪乳、佈比卡因和線粒體通透性轉換孔開放劑蒼術苷,終濃度分彆為1％、1 mmol/L和30 μmol/L,均孵育24 h.孵育結束後,採用Western blot法測定Bcl-2、Bax、燐痠化Bad (p-Bad)、caspase-3、活化的caspase-3、caspase-9、活化的caspase-9和細胞色素c(Cyt c)的錶達水平,採用RT-PCR法檢測Bcl-2 mRNA、Bax mRNA、Bad mRNA、caspase-9 mRNA和Cyt c mRNA的錶達水平.結果 與C組比較,B組Bax/Bcl-2比值、活化的caspase-3/caspase-3比值、活化的caspase-9/caspase-9比值和Bax mRNA/Bcl-2 mRNA比值升高,p-Bad錶達下調,Cyt c、Bad mRNA、caspase-9mRNA和Cyt c mRNA的錶達上調(P＜0.05),LB組上述各指標差異無統計學意義(P＞0.05);與B組比較,LB組和LBA組Bax/Bcl-2比值、活化的caspase-3/caspase-3比值和活化、caspase-9/caspase-9比值和Bax mRNA/Bcl-2 mRNA比值降低,p-Bad錶達上調,Cyt c、Bad mRNA、caspase-9 mRNA和Cyt cmRNA的錶達下調(P＜0.05);與LB組比較,LBA組Bax/Bcl-2比值、活化的caspase-3/caspase-3比值、活化的caspase-9/caspase-9比值和Bax mRNA/Bcl-2 mRNA比值升高,p-Bad錶達下調,Cyt c、BadmRNA、caspase-9 mRNA和Cyt c mRNA的錶達上調(P＜0.05).結論 脂肪乳逆轉佈比卡因大鼠心肌毒性作用的機製與抑製線粒體通透性轉換孔的開放有關.
목적 평개선립체통투성전운공재지방유역전포비잡인대서심기독성중적작용.방법 배양H9c2심기세포,이105개/ml적밀도접충우6공판,채용수궤수자표법분위4조,매조6공,매공2 ml:대조조(C조)가입PBS 100μl;포비잡인조(B조)가입포비잡인,종농도위1 mmol/L;지방유+포비잡인조(LB조)동시가입지방유화포비잡인,종농도분별위1％화1 mmol/L;지방유+포비잡인+창술감조(LBA조)동시가입지방유、포비잡인화선립체통투성전환공개방제창술감,종농도분별위1％、1 mmol/L화30 μmol/L,균부육24 h.부육결속후,채용Western blot법측정Bcl-2、Bax、린산화Bad (p-Bad)、caspase-3、활화적caspase-3、caspase-9、활화적caspase-9화세포색소c(Cyt c)적표체수평,채용RT-PCR법검측Bcl-2 mRNA、Bax mRNA、Bad mRNA、caspase-9 mRNA화Cyt c mRNA적표체수평.결과 여C조비교,B조Bax/Bcl-2비치、활화적caspase-3/caspase-3비치、활화적caspase-9/caspase-9비치화Bax mRNA/Bcl-2 mRNA비치승고,p-Bad표체하조,Cyt c、Bad mRNA、caspase-9mRNA화Cyt c mRNA적표체상조(P＜0.05),LB조상술각지표차이무통계학의의(P＞0.05);여B조비교,LB조화LBA조Bax/Bcl-2비치、활화적caspase-3/caspase-3비치화활화、caspase-9/caspase-9비치화Bax mRNA/Bcl-2 mRNA비치강저,p-Bad표체상조,Cyt c、Bad mRNA、caspase-9 mRNA화Cyt cmRNA적표체하조(P＜0.05);여LB조비교,LBA조Bax/Bcl-2비치、활화적caspase-3/caspase-3비치、활화적caspase-9/caspase-9비치화Bax mRNA/Bcl-2 mRNA비치승고,p-Bad표체하조,Cyt c、BadmRNA、caspase-9 mRNA화Cyt c mRNA적표체상조(P＜0.05).결론 지방유역전포비잡인대서심기독성작용적궤제여억제선립체통투성전환공적개방유관.
Objective To evaluate the effect of mitochondrial permeability transition pore (mPTP) in lipid emulsion-induced inversion of bupivacaine myocardiotoxicity in rats.Methods H9c2 cells were inoculated in 6-well plates at a density of 105 cells/ml, and were randomly divided into 4 groups (6 wells in each group, 2 ml/well) using a random number table: control group (group C) , bupivacaine group (group B) , lipid emusion + bupivacaine group (group LB) , and lipid emusion + bupivacaine + atractyloside group (group LBA).Phosphate buffer solution 100 μl was added to the culture medium in group C.In group B, bupivacaine was added to the culture medium with the final concentration of 1 mmol/L.In group LB, lipid emusion and bupivacaine were added to the culture medium with the final concentrations of 1％ and 1 mmol/L, respectively.In group LBA, lipid emusion, bupivacaine and atractyloside (an mPTP opener) were added to the culture medium with the final concentrations of 1％, 1 mmol/L and 30 μmol/L, respectively.All the cells were incubated for 24 h.After the end of incubation, the expression of Bcl-2, Bax, phosphorylated Bad (p-Bad) , caspase-3, activated caspase-3, caspase-9,activated caspase-9 and cytochrome c (Cyt c) was detected using Western blot.The expression of Bcl-2 mRNA, Bax mRNA, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was detected using real-time reverse transcriptase polymerase chain reaction.The ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were calculated.Results Compared with group C,the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/ Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group B (P＜0.05) , and no significant change was found in the parameters mentioned above in group LB (P＞0.05).Compared with group B, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly decreased, the expression of p-Bad was up-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was down-regulated in LB and LBA groups (P＜ 0.05).Compared with group LB, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group LBA (P ＜ 0.05).Conclusion The mechanism underlying lipid emulsioninduced inversion of bupivacaine myocardiotoxicity is related to inhibited mPTP opening in rats.
