右美托咪定对心脏手术小鼠术后疼痛和心肌细胞凋亡的影响及机制研究  

Dexmedetomidine reduces pain and inhibits apoptosis of myocardial cells through the PI3K/AKT signaling pathway in mice during cardiac surgery.

作　　者：苏鸿莉[1] 张天花 李玲霞[1] SU Hong-li;ZHANG Tian-hua;LI Ling-xia(Department of Anesthesiology,Affiliated Hospital of Yan'an University,Yan'an Shaanxi 716000,China;Department of Cardiology,Yan'an Hospital of Traditional Chinese Medicine,Yan'an Shaanxi 716000,China.)

机构地区：[1]延安大学附属医院麻醉科,陕西延安716000 [2]延安市中医医院心血管内科,陕西延安716000

出　　处：《临床和实验医学杂志》2021年第18期1932-1936,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省社会发展科技攻关项目(编号:2016SF-075)。

摘　　要：目的探讨右美托咪定(DEX)对小鼠心脏手术疼痛以及心肌保护的作用及其有关机制。方法取24只8周龄无特定病原体(SPF)雄性C57BL/6小鼠用于构建心肌缺血模型,并根据随机数字表法分为3组:右美托咪定低剂量组(25μg/kg)、高剂量组(50μg/kg)和模型组(注射等量生理盐水)。术后3 d,对小鼠进行疼痛评估。然后分离模型小鼠心肌细胞,根据实验目的分为:对照组:等体积磷酸盐缓冲溶液干预;渥曼青霉素(Wm)组:PI3K抑制剂渥曼青霉素(1 mg/mL)干预;DEX组:右美托咪定(10μmol/L)干预;DEX+Wm组:右美托咪定(10μmol/L)和渥曼青霉素(1 mg/mL)干预。培养72 h后,采用CCK-8检测细胞增殖、TUNEL检测凋亡情况;免疫印迹检测小鼠心肌细胞中磷脂酰肌醇-3-激酶(PI3K)、丝磷酸化苏氨酸蛋白激酶(p-AKT)、Caspase-3和Bcl-2的蛋白表达情况。结果与模型组比较,右美托咪定低剂量组和高剂量组可显著减轻心脏搭桥手术小鼠MGS得分(P<0.05),其中以右美托咪定高剂量组作用更明显(P<0.05);右美托咪定低剂量组和高剂量组小鼠的心率和平均动脉压在心脏搭桥手术后恢复显著(P<0.05)。右美托咪定干预12~72 h,与对照组相比,Wm组和DEX组心肌细胞活力显著降低(P<0.05),而DEX+Wm组与对照组相比则差异无统计学意义(P>0.05);与Wm组相比,DEX组和DEX+Wm组心肌细胞活力显著上升,且DEX+Wm组也显著高于DEX组(P<0.05)。与对照组相比,右美托咪定处理组小鼠心肌细胞凋亡率降低(P<0.05),而该作用被PI3K抑制剂渥曼青霉素阻断(P<0.05)。在右美托咪定处理的小鼠心肌细胞中,Bcl-2的表达水平上调,而Caspase-3的表达水平显著下调(P<0.05);上述作用同样被PI3K抑制剂渥曼青霉素阻断(P<0.05)。与对照组相比,DEX处理小鼠的心肌细胞内PI3K和AKT的表达水平明显升高,且AKT磷酸化也显著上调(P<0.05)。PI3K抑制剂Wm可明显拮抗右美托咪定的心肌细胞抗凋亡作用(P<0.05)。结论右美托咪�Objective To investigate the effect of dexmedetomidine(DEX)on the pain and myocardial protection of mouse cardiac surgery and its related mechanisms.Methods Twenty-four 8-week-old male C57BL/6 mice with Specific Pathogen Free(SPF)were used to construct the myocardial ischemia model and were divided into 3 groups according to the random number table method:Dexmedetomidine low-dose group(25μg/kg),high-dose group(50μg/kg)and model group(injected equal amount of saline).Three days after surgery,the mice were evaluated for pain.Then the model mouse cardiomyocytes were isolated.According to the purpose of the experiment,they were divided into:control group:equal volume PBS intervention;wortmannin(Wm)group:PI3K inhibitor wortmannin(1 mg/mL)intervention;DEX group:dexmedetomidine(10μmol/L)intervention;DEX+Wm group:dexmedetomidine(10μmol/L)and wortmannin(1 mg/mL)intervention.After 72 hours of culture,CCK-8 was used to detect cell proliferation and TUNEL was used to detect apoptosis.The protein expressions of phosphatei-dylinositol 3 kinase(PI3K),phosphorylation serine protein kinase(p-Akt),Caspase-3 and Bcl-2 in mouse cardiomyocytes were detected by Western blotting.Results Pretreatment with different doses of dexmedetomidine could significantly reduce the MGS scores of mice undergoing cardiac bypass surgery(P<0.05),and the effect of high-dose dexmedetomidine group was more obvious(P<0.05).The heart rate and mean arterial pressure of mice in the low-dose and high-dose groups of dexmedetomidine recovered significantly after cardiac bypass surgery(P<0.05).After dexmedetomidine intervention for 12 to 72 h,compared with the control group,the vitality of myocardial cells in the Wm group and the DEX group was significantly reduced(P<0.05),while the DEX+Wm group had no significant difference compared with the control group(P>0.05).Compared with the Wm group,the vitality of myocardial cells in the DEX group and the DEX+Wm group increased significantly,and the DEX+Wm group was also significantly higher than the DEX group(P<0.05

关 键 词：小鼠 右美托咪定 心肌保护 疼痛 凋亡 PI3K/AKT信号通路 

分 类 号：R614[医药卫生—麻醉学]