摘要
目的评价磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路在富氢水(HRS)减轻心肺转流(CPB)致大鼠心肌损伤中的作用。方法清洁级成年雄性SD大鼠40只,10~12周龄,体重350~400 g。采用随机数字表法分为四组:假手术组(S组)、CPB组(C组)、HRS处理组(H组)和PI3K抑制剂组(P组),每组10只。S组仅进行血管穿刺置管,置管后通过尾静脉注射生理盐水6 ml/kg,机械通气90 min,其余各组建立无血预充心脏不停跳CPB模型,CPB 60 min。CPB前60 min,P组通过腹腔内注射PI3K抑制剂6 ml/kg(LY294002,5μg/ml)。CPB前30 min,C组通过尾静脉注射生理盐水6 ml/kg,H组和P组通过尾静脉注射HRS 6 ml/kg。四组均于CPB停机2 h后取相应标本进行检测。采用Masson染色法观察心肌组织形态学变化;测定大鼠心肌含水量;采用ELISA法测定血浆心肌损伤标记物[心肌肌钙蛋白I(cTnI)、乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)]和炎性因子[白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]以及心肌组织氧化应激指标[丙二醛(MDA)、髓过氧化物酶(MPO)、氧化物歧化酶(SOD)]浓度;采用Western blot法检测心肌组织凋亡相关蛋白[B细胞淋巴瘤-2蛋白(Bcl-2)、Bcl-2相关x蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)]及PI3K信号通路相关蛋白[PI3K、Akt、磷酸化Akt(p-Akt)、血红素加氧酶-1(HO-1)]含量。结果S组心肌组织结构基本正常,C组心肌纤维化且明显损伤,H组心肌纤维损伤得到改善。与H组比较较,P组大鼠心肌损伤程度较为严重。与S组比较,C组、H组和P组心肌含水量、血浆cTnI、LDH、CK-MB和IL-1β、IL-6、TNF-α浓度、心肌组织MDA、MPO浓度、Bax、caspase-3蛋白含量明显升高(P<0.05),SOD浓度、Bcl-2和PI3K、p-Akt、HO-1蛋白含量明显降低(P<0.05)。与C组比较,H组心肌含水量、血浆cTnI、LDH、CK-MB和IL-1β、IL-6、TNF-α浓度、心肌组织MDA、MPO浓度、心肌组织Bax、caspase-3蛋白含量明显降低(P<0.05),心肌组织SOD浓度、Bcl-2和PI3K、p-Akt、HO-1蛋白含量明显升高(P<0.05)。与H组比较,P组心肌含水量、血浆cTnI、LDH、CK-MB和IL-1β、IL-6、TNF-α浓度、心肌组织MDA、MPO浓度、心肌组织Bax、caspase-3蛋白含量明显升高(P<0.05),心肌组织SOD浓度、心肌组织Bcl-2和PI3K、p-Akt、HO-1蛋白含量明显降低(P<0.05)。C组和P组上述指标差异无统计学意义。结论HRS减轻CPB致大鼠心肌损伤的机制与激活PI3K/Akt信号通路有关。
Objective To evaluate the role of phosphatidyl inositol 3 kinase/protein kinase B(PI3K/Akt)signaling pathway in HRS-induced reduction of cardiopulmonary bypass(CPB)-caused myocardial injury in rats.Methods Forty clean-grade adult male SD rats,10-12 weeks old,weighing 350-400 g were divided into four groups by random number table method(n=10):sham operation group(group S),CPB group(group C),HRS treatment group(group H)and PI3K inhibitor group(group P).Group S was placed arteriovenous catheter only without CPB and mechanically ventilated for 90 minutes,and after placing arteriovenous catheter normal saline 6 ml/kg was injected through tail vein.CPB mode was established for 60 minutes in other groups.60 minutes before CPB,the PI3K inhibitor 6 ml/kg(LY294002,5μg/ml)was injected intraperitoneally into their abdomen,30 minutes before CPB,normal saline 6 ml/kg was injected through the tail vein in group C,HRS 6 ml/kg was injected through tail vein in groups H and P.The corresponding specimens were taken for testing after the CPB was stopped for 2 hours in all four groups.Morphology of the myocardial tissues was observed by Masson staining.The myocardial water content was determined.The myocardial injury markers[cardiac troponin I(cTnI),lactate dehydrogenase(LDH),creatinine kianse-MB(CK-MB)],inflammatory factors[interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α)]in plasma and oxidative stress factors[malonaldehyde(MDA),myeloperoxidase(MPO),superoxidedimutase(SOD)]concentration levels were detected by ELISA.Apoptosis-related proteins for myocardial cells[B-cell lymphoma-2(Bcl-2),Bcl-2 associated x protein(Bax),cysteinyl aspartate specific proteinase-3(caspase-3)]and PI3K signaling pathway related proteins[PI3K,Akt,phosphorylated Akt(p-Akt),heme oxygenase-1(HO-1)]were detected by Western blot.Results Masson trichrome staining showed that myocardial tissue structure in group S was basically normal,myocardial fibrosis and seriously injury in group C,HRS treatment in group H improved myocardial fibrosis and injury,myocardial damage in group P was more serious than that in group H.Compared with group S,the myocardial water content,cTnI,LDH,CK-MB and IL-1β,IL-6,TNF-αin plasma,MDA,MPO,Bax,caspase-3 levels were significantly increased(P<0.05),SOD,Bcl-2 and PI3K,p-Akt,HO-1 levels were significantly decreased in groups C,H,and P(P<0.05).Compared with group C,the myocardial water content,cTnI,LDH,CK-MB and IL-1β,IL-6,TNF-αin plasma,MDA,MPO,Bax,caspase-3 levels were significantly decreased in group H(P<0.05),SOD,Bcl-2 and PI3K,p-Akt,HO-1 levels were significantly increased in group H(P<0.05).Compared with group H,the myocardial water content,cTnI,LDH,CK-MB,IL-1β,IL-6,TNF-αin plasma,MDA,MPO,Bax,caspase-3 levels were significantly increased in group P(P<0.05),SOD,Bcl-2 and PI3K,p-Akt,HO-1 levels were significantly decreased in group P(P<0.05).There were no significant differences in the above indexes between the group C and group P.Conclusion The role of HRS reducing CPB-caused myocardial injury is related to activating the PI3K/Akt signaling pathway in rats.
作者
崔波
宋丹丹
孙莹杰
CUI Bo;SONG Dandan;SUN Yingjie(Department of Anesthesiology,General Hospital of Northern Theater Command,Shenyang 110016,China)
出处
《临床麻醉学杂志》
CAS
CSCD
北大核心
2022年第5期520-525,共6页
Journal of Clinical Anesthesiology
基金
辽宁省重点研发计划项目(2020JH2/10300051)。
关键词
心肌损伤
心肺转流
富氢水
PI3K/AKT信号通路
Myocardial injury
Cardiopulmonary bypass
Hydrogen-rich solution
PI3K/Akt signaling pathway
